We report a complete 3D structural model of typical epithelial primary cilia based on structural maps of full-length primary cilia obtained by serial section electron tomography. Our data demonstrate ...the architecture of primary cilia differs extensively from the commonly acknowledged 9+0 paradigm. The axoneme structure is relatively stable but gradually evolves from base to tip with a decreasing number of microtubule complexes (MtCs) and a reducing diameter. The axonemal MtCs are cross-linked by previously unrecognized fibrous protein networks. Such an architecture explains why primary cilia can elastically withstand liquid flow for mechanosensing. The nine axonemal MtCs in a cilium are found to differ significantly in length indicating intraflagellar transport processes in primary cilia may be more complicated than that reported for motile cilia. The 3D maps of microtubule doublet–singlet transitions generally display longitudinal gaps at the inner junction between the A- and B-tubules, which indicates the inner junction protein is a major player in doublet–singlet transitions. In addition, vesicles releasing from kidney primary cilia were observed in the structural maps, supporting that ciliary vesicles budding may serve as ectosomes for cell–cell communication.
Centrioles are vital cellular structures that form centrosomes and cilia. The formation and function of cilia depends on a set of centriole's distal appendages. In this study, we use correlative ...super resolution and electron microscopy to precisely determine where distal appendage proteins localize in relation to the centriole microtubules and appendage electron densities. Here we characterize a novel distal appendage protein ANKRD26 and detail, in high resolution, the initial steps of distal appendage assembly. We further show that distal appendages undergo a dramatic ultra-structural reorganization before mitosis, during which they temporarily lose outer components, while inner components maintain a nine-fold organization. Finally, using electron tomography we reveal that mammalian distal appendages associate with two centriole microtubule triplets via an elaborate filamentous base and that they appear as almost radial finger-like protrusions. Our findings challenge the traditional portrayal of mammalian distal appendage as a pinwheel-like structure that is maintained throughout mitosis.
FUS-proteinopathies, a group of heterogeneous disorders including ALS-FUS and FTLD-FUS, are characterized by the formation of inclusion bodies containing the nuclear protein FUS in the affected ...patients. However, the underlying molecular and cellular defects remain unclear. Here we provide evidence for mitochondrial localization of FUS and its induction of mitochondrial damage. Remarkably, FTLD-FUS brain samples show increased FUS expression and mitochondrial defects. Biochemical and genetic data demonstrate that FUS interacts with a mitochondrial chaperonin, HSP60, and that FUS translocation to mitochondria is, at least in part, mediated by HSP60. Down-regulating HSP60 reduces mitochondrially localized FUS and partially rescues mitochondrial defects and neurodegenerative phenotypes caused by FUS expression in transgenic flies. This is the first report of direct mitochondrial targeting by a nuclear protein associated with neurodegeneration, suggesting that mitochondrial impairment may represent a critical event in different forms of FUS-proteinopathies and a common pathological feature for both ALS-FUS and FTLD-FUS. Our study offers a potential explanation for the highly heterogeneous nature and complex genetic presentation of different forms of FUS-proteinopathies. Our data also suggest that mitochondrial damage may be a target in future development of diagnostic and therapeutic tools for FUS-proteinopathies, a group of devastating neurodegenerative diseases.
Micro electrical discharge machining (micro EDM) is able to remove conductive material by non-contact instantaneous high temperature, which is more suitable for machining titanium and its alloys ...compared with traditional machining methods. To further improve the machining efficiency and machined surface quality of micro EDM, the nano particle surfactant mixed micro EDM method is put forward in this paper. Experiments were conducted to explore the effect of nano particle surfactant on the micro EDM performance of titanium alloy. The results show that the material removal rate of micro EDM in dielectric mixed with TiO2 is the highest when open-circuit voltage is 100 V, followed by Al2O3 and ZrO2. Lower tool wear rate can be produced by using dielectric mixed with nano particle surfactant. The taper ratio of micro EDM in dielectric mixed with nano particle surfactant is higher than that in deionized water. The surface roughness Ra of micro EDM in dielectric mixed with TiO2 can be 50% lower than that in deionized water. It is helpful to improve the machining performance by adding surface surfactant in the dielectric of micro EDM.
Fatigue performance is a serious concern for mechanical components subject to cyclical stresses, particularly where safety is paramount. The fatigue performance of components relies closely on their ...surface integrity because the fatigue cracks generally initiate from free surfaces. This paper reviewed the published data, which addressed the effects of machined surface integrity on the fatigue performance of metal workpieces. Limitations in existing studies and the future directions in anti-fatigue manufacturing field were proposed. The remarkable surface topography (e.g., low roughness and few local defects and inclusions) and large compressive residual stress are beneficial to fatigue performance. However, the indicators that describe the effects of surface topography and residual stress accurately need further study and exploration. The effect of residual stress relaxation under cycle loadings needs to be precisely modeled precisely. The effect of work hardening on fatigue performance had two aspects. Work hardening could increase the material yield strength, thereby delaying crack nucleation. However, increased brittleness could accelerate crack propagation. Thus, finding the effective control mechanism and method of work hardening is urgently needed to enhance the fatigue performance of machined components. The machining-induced metallurgical structure changes, such as white layer, grain refinement, dislocation, and martensitic transformation affect the fatigue performance of a workpiece significantly. However, the unified and exact conclusion needs to be investigated deeply. Finally, different surface integrity factors had complicated reciprocal effects on fatigue performance. As such, studying the comprehensive influence of surface integrity further and establishing the reliable prediction model of workpiece fatigue performance are meaningful for improving reliability of components and reducing test cost.
GaAs monolithic microwave integrated circuits (MMICs) with different back metallization systems (TiW/Au and Au/Ti/Au) exhibit different problems in the automatic Au-Sn eutectic bonding process, such ...as edge breakage or excessive voids. In this study, the formation mechanism of the edge breakage and excessive voids were investigated to prevent the damage of the MMICs in mass production scenarios. The microstructure and elemental distribution were studied using a scanning electron microscope and energy-dispersive spectroscopy. The void contents of the brazed region were measured with three-dimensional computed tomography. The top Au layer of the TiW/Au metallization partially dissolved in the melting An-Sn solder. Consequently, liquidus temperature of the solder increased, leading to isothermal solidification with the formation of ζ-Au
Sn in the scrubbing process, which was the reason for the edge breakage. The terminal Au film of the Au/Ti/Au metallization completely dissolved in the melting An-Sn solder. The metallurgical combination was achieved by the formation of the TiAu
intermetallic compound between the Au-Sn solder and the Ti layer. The wettability of Au-Sn solder on Ti layer should be improved to prevent the formation of the excessive voids.
Aggregation behaviors in mixtures of an anionic gemini surfactant 1,3-bis(N-dodecyl-N-propanesulfonate sodium)–propane (C12C3C12(SO3)2) and a cationic single-chain surfactant cetyltrimethylammonium ...bromide (CTAB) have been investigated in aqueous solutions at pH 9.5 by turbidity, rheology, isothermal titration microcalorimetry (ITC), cryogenic transmission electron microscopy, and dynamic light scattering. Reversible aggregate transitions from spherical micelles to wormlike micelles, vesicles, and back to wormlike micelles and spherical micelles are successfully realized through fine regulation over the mixing ratio of surfactants, i.e., the anionic/cationic charge ratio. The five aggregate regions display distinguished phase boundaries so that the aggregate regions can be well controlled. From thermodynamic aspect, the ITC curves clearly reflect all the aggregate transitions and the related interaction mechanism. The self-assembling ability of the C12C3C12(SO3)2/CTAB mixtures are significantly improved compared with both individual surfactants. Micelle growth from spherical to long wormlike micelles takes place at a relative low total concentration, i.e., 2.0 mM. The wormlike micelle solution at 10 mM or higher shows high viscosity and shear thinning property. Moreover, the C12C3C12(SO3)2/CTAB mixtures do not precipitate even at 1:1 charge ratio and relative high concentration. It suggests that applying gemini surfactant should be an effective approach to improve the solubility of anionic/cationic surfactant mixtures and in turn may promote applications of the surfactant mixtures.
A complex modulation algorithm for focusing beam shaping with a phase-only spatial light modulator is designed. This method modulates the amplitude and phase of a collimated beam synchronously, and ...the modulated beam passing through an objective lens will generate a shaped focusing beam. The characteristic of the complex modulation was studied, while the Mixed-Region Amplitude Freedom (MRAF) method was selected as a comparative reference in this research. In the theoretical simulation, the complex modulation shows excellent performance with a roughness of 0.32% RMS and 0.54% MAX. Otherwise, to further verify this complex modulation algorithm, an experiment was implemented to generate a square-shaped focusing flattop beam. The complex modulation generated a flattop beam with a roughness of 3.1% RMS and 6.1% MAX, better than the MRAF method. This result also proves that this complex modulation has better robustness.
Exocysts are highly conserved octameric complexes that play an essential role in the tethering of Golgi-derived vesicles to target membranes in eukaryotic organisms. Genes encoding the EXO70 subunit ...are highly duplicated in plants. Based on expression analyses, we proposed previously that individual EXO70 members may provide the exocyst with functional specificity to regulate cell type— or cargo-specific exocytosis, although direct evidence is not available. Here, we show that, as a gene expressed primarily during tracheary element (TE) development, EXO70A1 regulates vesicle trafficking in TE differentiation in Arabidopsis thaliana. Mutations of EXO70A1 led to aberrant xylem development, producing dwarfed and nearly sterile plants with very low fertility, reduced cell expansion, and decreased water potential and hydraulic transport. Grafting of a mutant shoot onto wild-type rootstock rescued most of these aboveground phenotypes, while grafting of a wild-type shoot to the mutant rootstock did not rescue the short root hair phenotype, consistent with the role of TEs in hydraulic transport from roots to shoots. Histological analyses revealed an altered pattern of secondary cell wall thickening and accumulation of large membrane-bound compartments specifically in developing TEs of the mutant. We thus propose that EXO70A1 functions in vesicle trafficking in TEs to regulate patterned secondary cell wall thickening.
Abstract
We aimed to elucidate the differences in genomic methylation patterns between ADLI and non-ADLI patients to identify DNA methylation-based biomarkers. Genome-wide DNA methylation patterns ...were obtained using Infinium MethylationEPIC (EPIC) BeadChip array to analyze 14 peripheral blood samples (7 ADLI cases, 7 non-ADLI controls). Changes in the mRNA and DNA methylation in the target genes of another 120 peripheral blood samples (60 ADLI cases, 60 non-ADLI controls) were analyzed by real-time polymerase chain reaction and pyrosequencing, respectively. A total of 308 hypermethylated CpG sites and 498 hypomethylated CpG sites were identified. Significantly, hypermethylated CpG sites cg06961147 and cg24666046 in TANC1 associated with ADLI was identified by genome-wide DNA methylation profiling. The mRNA expression of TANC1 was lower in the cases compared to the controls. Pyrosequencing validated these two differentially methylated loci, which was consistent with the results from the EPIC BeadChip array. Receiver operating characteristic analysis indicated that the area under the curve of TANC1 (cg06961147, cg24666046, and their combinations) was 0.812, 0.842, and 0.857, respectively. These results indicate that patients with ADLI have different genomic methylation patterns than patients without ADLI. The hypermethylated differentially methylated site cg06961147 combined with cg24666046 in TANC1 provides evidence for the diagnosis of ADLI.
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