A single N-terminal phosphomimic disrupts TDP-43 polymerization, phase separation, and RNA splicing.

TDP-43 is an RNA-binding protein active in splicing that concentrates into membraneless ribonucleoprotein granules and forms aggregates in amyotrophic lateral sclerosis (ALS) and Alzheimer's disease. Although best known for its predominantly disordered C-terminal domain which mediates ALS inclusions, TDP-43 has a globular N-terminal domain (NTD). Here, we show that TDP-43 NTD assembles into head-to-tail linear chains and that phosphomimetic substitution at S48 disrupts TDP-43 polymeric assembly, discourages liquid-liquid phase separation (LLPS) in vitro, fluidizes liquid-liquid phase separated nuclear TDP-43 reporter constructs in cells, and disrupts RNA splicing activity. Finally, we present the solution NMR structure of a head-to-tail NTD dimer comprised of two engineered variants that allow saturation of the native polymerization interface while disrupting higher-order polymerization. These data provide structural detail for the established mechanistic role of the well-folded TDP-43 NTD in splicing and link this function to LLPS. In addition, the fusion-tag solubilized, recombinant form of TDP-43 full-length protein developed here will enable future phase separation and in vitro biochemical assays on TDP-43 function and interactions that have been hampered in the past by TDP-43 aggregation.

Creation of a 3-dimensional virtual dental patient for computer-guided surgery and CAD-CAM interim complete removable and fixed dental prostheses: A clinical report.

This clinical report proposes a digital workflow using 2-dimensional (2D) digital photographs, a 3D extraoral facial scan, and cone beam computed tomography (CBCT) volumetric data to create a 3D virtual patient with craniofacial hard tissue, remaining dentition (including surrounding intraoral soft tissue), and the realistic appearance of facial soft tissue at an exaggerated smile under static conditions. The 3D virtual patient was used to assist the virtual diagnostic tooth arrangement process, providing patient with a pleasing preoperative virtual smile design that harmonized with facial features. The 3D virtual patient was also used to gain patient's pretreatment approval (as a communication tool), design a prosthetically driven surgical plan for computer-guided implant surgery, and fabricate the computer-aided design and computer-aided manufacturing (CAD-CAM) interim prostheses.

Trophic transfer of DDE, BP-3 and chlorpyrifos from microplastics to tissues in Dicentrarchus labrax.

Microplastic pollution and associated chemical contaminants is a topic of growing interest. In recent years, the number of publications reporting the presence of microplastics (MPs) in marine organisms has increased exponentially. However, there is a gap in knowledge about the trophic transfer of contaminants from microplastics to animal tissues, as well as possible health effects. In this study we analyzed the trophic transfer and biomagnification of three chemical pollutants present in microplastics: dichlorodiphenyldichloroethylene (DDE-p,p'), benzophenone 3 (BP-3) and chlorpyrifos (CPS). The reference values used were concentrations found in environmental microplastics in the Canary Islands (minimum and maximum). European seabass (Dicentrarchus labrax) were fed for 60 days with 5 different treatments: A) feed; B) feed with chemical pollutants at maximum concentration; C) feed + 10 % virgin MPs; D) feed + 10 % MPs with chemical pollutants at minimum concentration; E) feed + 10 % MPs with chemical pollutants at maximum concentration. We detected trophic transfer of DDE-p,p', CPS and BP-3 from the feed (treatment B) to the muscle and liver of fish. In the case of DDE-p,p', transfer to liver and muscle was also observed in the treatments consisting of feed plus plastics with different levels of contamination (C, D and E). No effect of the experimental treatments on fish condition indices was observed.

Bioaccumulation of additives and chemical contaminants from environmental microplastics in European seabass (Dicentrarchus labrax).

Marine microplastic pollution is one of the biggest environmental problems we face. The growth of plastic production has not ceased since the 1950s and it is currently estimated that 368 tons of plastic were produced in 2019 (PlascticsEurope, 2020). Geyer et al. (2017) estimate that 79% of the plastic produced in the world still remains in the environment; this plastic due to the effect of degradation and subsequent fragmentation, is present in the form of microplastics in all oceans and, due to its small size can be ingested by fish and filter-feeding organisms. In addition, microplastics have additives and chemical contaminants associated with them, and the potential effect of microplastic ingestion on marine organisms, and through them, the potential risk to humans, is unknown. In the present study, European seabass (Dicentrarchus labrax) were fed for 60 days with three treatments: Control (feed), MP (feed with 10% virgin microplastics) and EMP (feed with 10% environmental microplastics), being the first study to evaluate long-term accumulation of contaminants due to ingestion of environmental microplastics (EMP) in fish. Both plastic additives such as PBDEs, and chemical contaminants adsorbed from the environment such as PCBs and DDE, were analyzed in the EMP, feed and liver. The concentration of microplastics in the feed was calculated based on the MPs/zooplankton wet weight (WW) ratio of 0.1 found in an area of maximum accumulation in the Canary Islands. Therefore, it is an experiment that simulates real conditions, but in the worst-case scenario, using both, concentrations based on data obtained in oceanographic campaigns and microplastics collected from the environment. Our results show that in this scenario, additives and chemical contaminants adsorbed on EMPs bioaccumulate in fish liver due to long-term ingestion of microplastics.

Impact of polypropylene microplastics and chemical pollutants on European sea bass (Dicentrarchus labrax) gut microbiota and health.

Plastic pollution has become a global problem for marine ecosystems. Microplastics (MPs) are consumed by several marine organisms, including benthic and pelagic fish species that confuse them with food sources, thus contributing to bioaccumulation along the food chain. In addition to structural intestinal damage, ingestion of MPs represents a pathway for fish exposure to potentially hazardous chemicals, too. Most of them are endocrine disrupters, genotoxic or induce immune depression in fish. Accordingly, we assessed the combined toxicological effects of microplastics (MPs) and adsorbed pollutants by adding them to marine fish diet. European sea bass (Dicentrarchus labrax) juveniles were fed for 60 days with feeds containing polypropylene MPs, either virgin or contaminated with chemical pollutants (a blend of dichlorodiphenyldichloroethylene, chlorpyrifos, and benzophenone-3). The data demonstrated a synergic action of MPs and chemical pollutants to induce an inflammatory-like response in distal intestine of sea bass as shown by the up regulation of cytokine il-6 and tnf-α expression. Morphological analysis detected the presence of a focus of lymphocytes in anterior and posterior intestinal segments of fish fed with contaminants in the diet. With regard to microbiota, significant changes in bacterial species richness, beta diversity, and composition of gut microbiota were observed as a consequence of both pollutants and polluted MPs ingestion. These perturbations in gut microbial communities, including the reduction of beneficial lactic acid bacteria and the increase in potential pathogenic microorganism (Proteobacteria and Vibrionales), were undeniable signs of intestinal dysbiosis, which in turn confirmed the signs of inflammation caused by pollutants, especially when combined with MPs. The results obtained in this study provide, therefore, new insights into the potential risks of ingesting MPs as pollutant carriers in marine fish.

Key nutritional factors and interactions during larval development of pikeperch (Sander lucioperca).

The effects of 8 nutritional variables (Ca/P, Eicosapentaenoic acid (20:5n-3) + Docosahexaenoic acid (22:6n - 3) (EPA + DHA), Arachidonic acid (20:4n - 6) (ARA), Se, vitamins E, C, D and A) were investigated to identify their respective importance and interactions in pikeperch larval development. In this respect, two modalities (low and high levels) of each variable were tested through a fractional factorial experimental design allowing a reduction from 256 (28) to 16 (28 - 4) experimental units. Survival was significantly higher in larvae fed a high Ca/P diet while larval growth was significantly lower in larvae fed the same diet variant, associated with a higher incidence of kyphosis and pectoral anomalies in these larvae. Lordosis and scoliosis seemed to be mostly affected by dietary long chain polyunsaturated fatty acids (LC-PUFAs). A significant interaction was shown between n-3 LC-PUFA and vitamin C on jaw anomalies, while myocyte-specific enhancer factor 2C (mef2c) gene expression correlated positively with dietary vitamin C increment. Results also demonstrated an effect of the different nutrients and their interactions on the activity levels of digestive enzymatic activities. The results of the present study highlight the importance of the interactions between Ca/P, LC-PUFAs and vitamins C and E, suggesting their essential roles as key nutritional factors influencing pikeperch larval development.