The C-Terminal TDP-43 Fragments Have a High Aggregation Propensity and Harm Neurons by a Dominant-Negative Mechanism

TAR DNA binding protein 43 KD (TDP-43) is an essential gene that regulates gene transcription, mRNA splicing and stability. In amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD), two fatal neurodegenerative diseases, TDP-43 is fragmented, generating multiple fragments that include the C-terminal fragment of ∼25 KD. The role of these fragments in the pathogenesis of ALS and FTD is not clear. Here we investigated the aggregation propensity in various polypeptide regions of TDP-43 in mammalian cells and the effect of these fragments on cultured neurons. By expressing the full length and various TDP-43 fragments in motor neuron-derived NSC-34 cells and primary neurons, we found that both N- and C-terminal fragments of TDP-43 are prone to aggregate and the C-terminal end of RRM2 region is required, though not sufficient, for aggregation. The aggregation of the TDP-43 fragments can drive co-aggregation with the full-length TDP-43, consequently reducing the nuclear TDP-43. In addition, the TDP-43 fragments can impair neurite growth during neuronal differentiation. Importantly, overexpression of the full-length TDP-43 rescues the neurite growth phenotype whereas knockdown of the endogenous TDP-43 reproduces this phenotype. These results suggest that TDP-43 fragments, particularly the pathologically relevant C-terminal fragments, can impair neuronal differentiation by dominant-negatively interfering with the function of the full length TDP-43, thus playing a role in pathogenesis in ALS and FTD

Glycosaminoglycans and other sulphated polysaccharides in calculogenesis of urinary stones

Naturally occurring glycosaminoglycans (GAGs) and other, semisynthetic, sulphated polysaccharides are thought to play an important role in urolithiasis. Processes involved in urinary stone formation are crystallization and crystal retention. Oxalate transport and renal tubular cell injury are determining factors in these processes. In this article experimental results concerning the possible mechanisms of action of GAGs and other sulphated polysaccharides are reviewed. GAGs are inhibitors of crystal growth and agglomeration and possibly also of nucleation. They can prevent crystal adherence, correct an abnormal oxalate flux and prevent renal tubular cell damage

Oxalate-induced ceramide accumulation in Madin-Darby canine kidney and LLC-PK1 cells

BACKGROUND: Oxalate exposure produces oxidant stress in renal epithelial cells leading to death of some cells and adaptation of others. The pathways involved in these diverse actions remain unclear, but appear to involve activation of phospholipase A2 (PLA2) and redistribution of membrane phospholipids. The present studies examined the possibility that oxalate actions may also involve increased accumulation of ceramide, a lipid-signaling molecule implicated in a variety of pathways, including those leading to apoptotic cell death. METHODS: Ceramide accumulation was examined in renal epithelial cells from pig kidney (LLC-PK1 cells) and from dog kidney [Madin-Darby canine kidney (MDCK cells)] using the diacylglycerol kinase assay. Sphingomyelin degradation was assessed by monitoring the disappearance of 3H-sphingomyelin from cells that had been prelabeled with [3H]-choline. The effects of oxalate were compared with those of other oxidants (peroxide, xanthine/xanthine oxidase), other organic acids (formate and citrate), and a known activator of sphingomyelinase in these cells [tumor necrosis factor-alpha (TNF-alpha)]. Separate studies determined whether oxalate-induced accumulation of ceramide could be blocked by pretreatment with antioxidants [Mn (III) tetrakis (1-methyl-4-pyridyl) porphyrin (Mn TMPyP, a superoxide dismutase mimetic) or N-acetylcysteine (NAC; an antioxidant)], with an inhibitor of ceramide synthase [fumonisin B1 (FB1)] or with an inhibitor of PLA2 [arachidonyl trifluoromethylketone (AACOCF3)]. RESULTS: Oxalate exposure produced a significant time- and concentration-dependent increase in cellular ceramide. A reciprocal decrease in 3H-sphingomyelin was observed under these conditions. Increases in cellular ceramide levels were also observed after treatment with other oxidants (hydrogen peroxide, and xanthine/xanthine oxidase), activators of sphingomyelinase (TNF-alpha), exogenous sphingomyelinase, or arachidonic acid. Formate produced similar (albeit smaller) effects, and citrate did not. The oxidant-induced increases in ceramide were attenuated by pretreatment with NAC (a glutathione precursor) and MnTMPyP (a superoxide dismutase mimetic), suggesting a role for cellular redox states. The oxalate-induced increase in ceramide was also attenuated by pretreatment with AACOCF3, suggesting a role for PLA2. Pretreatment with FB1 produced a small but statistically insignificant attenuation of the response to oxalate. CONCLUSIONS: Oxalate exposure produces a marked accumulation of ceramide in renal epithelial cells by a process that is redox sensitive and mediated in part by activation of PLA2. Since cellular sphingomyelin decreased as ceramide increased, it seems likely that oxalate actions are mediated, at least in part, by an increase in sphingomyelinase activity, although alterations in ceramide synthase are also possible. Further study is required to define the steps involved in oxalate actions and to determine the extent to which ceramide signaling mediates oxalate actions

The Design and Implementation of Virtual Roaming in Yunnan Diqing Tibetan traditional Villages

Traditional residence is the continuation of intangible cultural heritage and the primitive soil for development. At present, the protection and inheritance of traditional villages have been impacted by the process of modernization, and the phenomenon of assimilation is very serious. This article takes the above questions as the breakthrough point, and then analyzes why and how to use virtual reality technology to better solve the above problems, and take the Yunnan Diqing Tibetan traditional dwellings as the specific example to explore. First, using VR technology, with real images and sound, the paper simulate a near real virtual world. Secondly, we collect a large amount of real image information, and make the visualization model of building by using 3DMAX software platform, UV Mapping and Rendering optimization. Finally, the Vizard virtual reality development platform was used to establish the roaming system and realize the virtual interaction. The roaming system was posted online so that overcome the disadvantages of not intuitive and low capability of interaction, and these new ideas can give a whole new meaning in the protection projects of the cultural relic buildings. At the same time, visitors could enjoy the "Dian-style" architectural style and cultural connotation of dwelling house in Diqing Yunnan

The Design and Implementation of Virtual Roaming in Yunnan Diqing Tibetan traditional Villages

Traditional residence is the continuation of intangible cultural heritage and the primitive soil for development. At present, the protection and inheritance of traditional villages have been impacted by the process of modernization, and the phenomenon of assimilation is very serious. This article takes the above questions as the breakthrough point, and then analyzes why and how to use virtual reality technology to better solve the above problems, and take the Yunnan Diqing Tibetan traditional dwellings as the specific example to explore. First, using VR technology, with real images and sound, the paper simulate a near real virtual world. Secondly, we collect a large amount of real image information, and make the visualization model of building by using 3DMAX software platform, UV Mapping and Rendering optimization. Finally, the Vizard virtual reality development platform was used to establish the roaming system and realize the virtual interaction. The roaming system was posted online so that overcome the disadvantages of not intuitive and low capability of interaction, and these new ideas can give a whole new meaning in the protection projects of the cultural relic buildings. At the same time, visitors could enjoy the "Dian-style" architectural style and cultural connotation of dwelling house in Diqing Yunnan