The plasma membrane ATP release channel pannexin 1 has been implicated in numerous physiological and pathophysiological processes associated with purinergic signaling, including cancer progression, apoptotic cell clearance, inflammation, blood pressure regulation, oocyte development, epilepsy and neuropathic pain. Here, we present nearatomic resolution structures of Xenopus tropicalis and Homo sapiens PANX1 determined by cryo-electron microscopy that reveal a heptameric channel architecture. Compatible with ATP permeation, the transmembrane pore and cytoplasmic vestibule are exceptionally wide. An extracellular tryptophan ring located at the outer pore creates a constriction site, potentially functioning as a molecular sieve that restricts the size of permeable substrates. In combination with functional characterization, this work elucidates the previously unknown architecture of pannexin channels and establishes a foundation for understanding their unique channel properties as well as for developing rational therapies..

No abstract

BackgroundTissue engineering has emerged as a promising alternative for small-diameter vascular grafts. The aim of this study was to determine the feasibility of using decellularized aortae of fetal pigs (DAFPs) to construct tissue-engineered, small-diameter vascular grafts and to test the performance and application of DAFPs as vascular tissue-engineered scaffolds in the canine arterial system.MethodsDAFPs were prepared by continuous enzymatic digestion. Canine vascular endothelial cells (ECs) were seeded onto DAFPs in vitro and then the vascular grafts were cultured in a custom-designed vascular bioreactor system for 7 days of dynamic culture following 3 days of static culture. The grafts were then transplanted into the common carotid artery of the same seven dogs from which ECs had been derived (two grafts were prepared for each dog with one as a backup; therefore, a total of 14 tissue-engineered blood vessels were prepared). At 1, 3, and 6 months post-transplantation, ultrasonography and contrast-enhanced computed tomography (CT) were used to check the patency of the grafts. Additionally, vascular grafts were sampled for histological and electron microscopic examination.ResultsTissue-engineered, small-diameter vascular grafts can be successfully constructed using DAFPs and canine vascular ECs. Ultrasonographic and CT test results confirmed that implanted vascular grafts displayed good patency with no obvious thrombi. Six months after implantation, the grafts had been remodeled and exhibited a similar structure to normal arteries. Immunohistochemical staining showed that cells had evenly infiltrated the tunica media and were identified as muscular fibroblasts. Scanning electron microscopy showed that the graft possessed a complete cell layer, and the internal cells of the graft were confirmed to be ECs by transmission electron microscopy.ConclusionsTissue-engineered, small-diameter vascular grafts constructed using DAFPs and canine vascular ECs can be successfully transplanted to replace the canine common carotid artery. This investigation potentially paves the way for solving a problem of considerable clinical need, i.e., the requirement for small-diameter vascular grafts.

Stereolithography is an attractive technique for the fabrication of complex‐shaped ceramic components with high dimensional accuracy. One of the challenges in this technology is the development of high solid loading, low viscosity photosensitive ceramic suspension. In this study, the dispersion of zirconia in photocurable resin and the slurry properties were intensively investigated. Rheological measurements showed that DISPERBYK‐103 proved to be an effective dispersant. 42 vol% ZrO2 suspension was successfully prepared using 3.5 wt% DISPERBYK‐103 as the dispersant, with a suitable viscosity (4.88 Pa·s) below the maximum allowable viscosity value (5 Pa·s) for stereolithography applications. The adsorption behavior of DISPERBYK‐103 on the surface of zirconia powders was characterized by TG and FT‐IR, confirming the dispersion effect of dispersant. Contact angle measurements were also conducted to show that the adsorption of DISPERBYK‐103 could help to improve the wettability between powder and photocurable resin. Results showed that DISPERBYK‐103 was effective for the preparation of suitable slurries for the development of ZrO2 ceramics through stereolithography.

The cyanobacterial type I NAD(P)H dehydrogenase (NDH-1) complexes play a crucial role in a variety of bioenergetic reactions such as respiration, CO2 uptake, and cyclic electron transport around photosystem I. Two types of NDH-1 complexes, NDH-1MS and NDH-1MS′, are involved in the CO2 uptake system. However, the composition and function of the complexes still remain largely unknown. Here, we found that deletion of ndhM caused inactivation of NDH-1-dependent cyclic electron transport around photosystem I and abolishment of CO2 uptake, resulting in a lethal phenotype under air CO2 condition. The mutation of NdhM abolished the accumulation of the hydrophilic subunits of the NDH-1, such as NdhH, NdhI, NdhJ, and NdhK, in the thylakoid membrane, resulting in disassembly of NDH-1MS and NDH-1MS′ as well as NDH-1L. In contrast, the accumulation of the hydrophobic subunits was not affected in the absence of NdhM. In the cytoplasm, the NDH-1 subcomplex assembly intermediates including NdhH and NdhK were seriously affected in the ΔndhM mutant but not in the NdhI-deleted mutant ΔndhI. In vitro protein interaction analysis demonstrated that NdhM interacts with NdhK, NdhH, NdhI, and NdhJ but not with other hydrophilic subunits of the NDH-1 complex. These results suggest that NdhM localizes in the hydrophilic subcomplex of NDH-1 complexes as a core subunit and is essential for the function of NDH-1MS and NDH-1MS′ involved in CO2 uptake in Synechocystis sp. strain PCC 6803.

Zirconia ceramics have shown a wide applicable prospect in dental prosthetics because it possesses excellent mechanical performance and biocompatibility. The rheological behavior and curing properties of zirconia stereolithography slurry were studied, as well as the microstructure of the green, pyrolyzed, and sintered body.Variquat CC-42 NS was proved to be effective for zirconia powder dispersing in photocurable resin (SP-RC700). The curing characteristic of slurry showed that the depth of penetration tended to increase with the solid loading. Finally, the zirconia all-ceramic crowns were fabricated via DLP-based stereolithography printer. Results showed that the particles were evenly distributed in the cured resin matrix without obvious agglomeration, and the interlayered structure disappeared after binder burnout. The sample with a major crystalline phase of tetragonal zirconia and a porosity of 10(1)% were obtained after sintering at 1550°C. K E Y W O R D Sceramic stereolithography, microstructure, rheological properties, zirconia all-ceramic crown

The large size complex of cyanobacterial NAD(P)H dehydrogenase (NDH-1) complex (NDH-1L) plays crucial role in a variety of bioenergetic reactions such as respiration and cyclic electron flow around photosystem I. Although the complex has been isolated and identified, its biochemical function still remains to be clarified. Here, we highly purified the NDH-1L complex from the cells of Thermosynechococcus elongatus by Ni(2+) affinity chromatography and size-exclusion chromatography. The purified NDH-1L complex has an apparent total molecular mass of approximately 500 kDa. 14 known subunits were identified by mass spectrometry and immunoblotting, including the NdhS subunit containing ferredoxin (Fd)-docking site domain. Surface plasmon resonance measurement demonstrates that the NDH-1L complex could bind to Fd with the binding constant (K D) of 59 µM. Yeast two-hybrid system assay further confirmed the interaction of Fd with NdhS and indicated that NdhH is involved in the interaction. Our results provide direct biochemical evidence that the cyanobacterial NDH-1 complex catalyzes the electron transport from reduced Fd to plastoquinone via NdhS and NdhH.

No abstract