A Micro-Topography Measurement and Compensation Method for the Key Component Surface Based on White-Light Interferometry.

The grinding grooves of material removal machining and the residues of a machining tool on the key component surface cause surface stress concentration. Thus, it is critical to carry out precise measurements on the key component surface to evaluate the stress concentration. Based on white-light interferometry (WLI), we studied the measurement distortion caused by the reflected light from the steep side of the grinding groove being unable to return to the optical system for imaging. A threshold value was set to eliminate the distorted measurement points, and the cubic spline algorithm was used to interpolate the eliminated points for compensation. The compensation result agrees well with the atomic force microscope (AFM) measurement result. However, for residues on the surface, a practical method was established to obtain a microscopic 3D micro-topography point cloud and a super-depth-of-field fusion image simultaneously. Afterward, the semantic segmentation network U-net was adopted to identify the residues in the super-depth-of-field fusion image and achieved a recognition accuracy of 91.06% for residual identification. Residual feature information, including height, position, and size, was obtained by integrating the information from point clouds and super-depth-of-field fusion images. This work can provide foundational data to study surface stress concentration.

Cryo-EM structure of human heptameric pannexin 2 channel.

Pannexin 2 (Panx2) is a large-pore ATP-permeable channel with critical roles in various physiological processes, such as the inflammatory response, energy production and apoptosis. Its dysfunction is related to numerous pathological conditions including ischemic brain injury, glioma and glioblastoma multiforme. However, the working mechanism of Panx2 remains unclear. Here, we present the cryo-electron microscopy structure of human Panx2 at a resolution of 3.4 Å. Panx2 structure assembles as a heptamer, forming an exceptionally wide channel pore across the transmembrane and intracellular domains, which is compatible with ATP permeation. Comparing Panx2 with Panx1 structures in different states reveals that the Panx2 structure corresponds to an open channel state. A ring of seven arginine residues located at the extracellular entrance forms the narrowest site of the channel, which serves as the critical molecular filter controlling the permeation of substrate molecules. This is further verified by molecular dynamics simulations and ATP release assays. Our studies reveal the architecture of the Panx2 channel and provide insights into the molecular mechanism of its channel gating.

De Novo Design of Peptidic Positive Allosteric Modulators Targeting TRPV1 with Analgesic Effects.

Transient receptor potential vanilloid 1 (TRPV1) ion channel is a nociceptor critically involved in pain sensation. Direct blockade of TRPV1 exhibits significant analgesic effects but also incurs severe side effects such as hyperthermia, causing failures of TRPV1 inhibitors in clinical trials. In order to selectively target TRPV1 channels that are actively involved in pain-sensing, peptidic positive allosteric modulators (PAMs) based on the high-resolution structure of the TRPV1 intracellular ankyrin-repeat like domain are de novo designed. The hotspot centric approach is optimized for protein design; its usage in Rosetta increases the success rate in protein binder design. It is demonstrated experimentally, with a combination of fluorescence resonance energy transfer (FRET) imaging, surface plasmon resonance, and patch-clamp recording, that the designed PAMs bind to TRPV1 with nanomolar affinity and allosterically enhance its response to ligand activation as it is designed. It is further demonstrated that the designed PAM exhibits long-lasting in vivo analgesic effects in rats without changing their body temperature, suggesting that they have potentials for developing into novel analgesics.

Molecular mechanisms underlying menthol binding and activation of TRPM8 ion channel.

Menthol in mints elicits coolness sensation by selectively activating TRPM8 channel. Although structures of TRPM8 were determined in the apo and liganded states, the menthol-bounded state is unresolved. To understand how menthol activates the channel, we docked menthol to the channel and systematically validated our menthol binding models with thermodynamic mutant cycle analysis. We observed that menthol uses its hydroxyl group as a hand to specifically grab with R842, and its isopropyl group as legs to stand on I846 and L843. By imaging with fluorescent unnatural amino acid, we found that menthol binding induces wide-spread conformational rearrangements within the transmembrane domains. By Φ analysis based on single-channel recordings, we observed a temporal sequence of conformational changes in the S6 bundle crossing and the selectivity filter leading to channel activation. Therefore, our study suggested a 'grab and stand' mechanism of menthol binding and how menthol activates TRPM8 at the atomic level.

Evaluation of sampling sizes on the intertidal macroinfauna assessment in a subtropical mudflat of Hong Kong.

In this study, two types of sediment cores with different diameters were used to collect sediment samples from an intertidal mudflat in Hong Kong to investigate the influence of sampling unit on the quantitative assessment of benthic macroinfaunal communities. Both univariate and multivariate analyses were employed to detect differences in sampling efficiencies by the two samplers through total abundance and biomass, species richness and diversity, community structure, relative abundance of major taxa of the infaunal community. The species-area curves were further compared to find out the influence of the sampling units. Results showed that the two sampling devices provided similar information on the estimates of species diversity, density and species composition of the benthos in main part of the mudflat where the sediment was fine and homogenous; but at the station which contained coarse sand and gravels, the significant differences were detected between the quantitative assessments of macrobenthic infauna by the two samplers. Most importantly, the species-area curves indicated that more and smaller samples were better in capturing more species than less large ones when comparing an equal sampling area. Therefore, the efficiency of the sampler largely depended on the sediment properties, and sampling devices must be chosen based on the physical conditions and desired levels of precision on the organisms of the sampling program.

High incidence of plasmids in marine Vibrio species isolated from Mai Po Nature Reserve of Hong Kong.

Mai Po Nature Reserve is the largest mangrove ecosystem and the most polluted coastal water body in Hong Kong. Plasmids screening of 100 Vibrio isolates randomly showed 45 % of them contained 1-3 plasmids. These plasmid(s)-bearing isolates could be divided into 12 groups based on their plasmid profiles. Phylogenetic analysis of the partial 16S rRNA gene sequences confirmed that all plasmid(s)-bearing isolates belonged to Vibrio cholerae. Full DNA sequences of the plasmids in Groups I (pVCG1.1 and pVCG1.2), II (pVCG2.1), III (pVCG3.2) and IV (pVCG4.1) have been determined and the results showed that pVCG1.1, pVCG2.1 and pVCG3.2 were almost identical. Plasmids pVCG1.1, pVCG1.2 and pVCG4.1 are comprised of 4,439, 2,357 and 2,163 bp with the overall G+C content of 45.57, 53.54 and 43.09 %, respectively. pVCG1.1 is a novel plasmid, and plasmids pVCG1.2 and pVCG4.1 showed homology of replication initiation proteins to that of the theta type replicons. Attempts to cure the plasmids from their hosts were unsuccessful. These data suggest that plasmids of Vibrio spp. are a significant gene reservoir in the marine ecosystem.

Abundance, composition and vertical distribution of polycyclic aromatic hydrocarbons in sediments of the Mai Po Inner Deep Bay of Hong Kong.

The distribution and changes of polycyclic aromatic hydrocarbons (PAHs) contamination in mangrove sediments of Mai Po Inner Deep Bay Ramsar Site of Hong Kong SAR were investigated. Surface sediments (10 cm) collected from four sampling sites (SZ, SP, MF and M) exhibited significant spatial variations in concentrations of total PAH (with ΣPAHs ranging from 161.7 to 383.7 ng g(-1) dry wt), as well as the composition of 16 US EPA priority PAH compounds. The highest PAHs concentrations were found in the mangrove sediments. Moreover, a sediment core was extracted from mangrove area is used to reconstruct the high-resolution depositional record of PAHs by (210)Pb isotope analysis, showing the amounts of PAHs remained relatively constant for the past 41 years. Urbanization of Shenzhen Economic Zone, the rapid increase in vehicle numbers and energy consumption in the last two decades contributed to the PAHs detected in sediments. The source-diagnostic ratios indicated that pyrogenic input are important throughout the record and the surface sediments, and suggest that diesel fuel combustion, and hence traffic of heavier vehicles, is the most probable cause of PAHs.

Distribution and sources of polycyclic aromatic hydrocarbons in sediments of the Mai Po Inner Deep Bay Ramsar Site in Hong Kong.

The concentration of total polycyclic aromatic hydrocarbons (∑PAHs) and the 16 US EPA priority individual PAH compounds were analyzed in surface sediments from the Mai Po Inner Deep Bay, Ramsar Site of Hong Kong from December 2001 to Jun 2005, to investigate the spatial variability of anthropogenic pollutants. ∑PAHs concentrations ranged from 36.5 to 256.3 ng g(-1) dry weight with an average of 148.9 ng g(-1), comparable to other urbanized areas of the world, and there was little difference among different sampling times from December 2001 to June 2005. Based on comparison to the results from earlier study, it appears that a decrease of total PAHs concentration has occurred since 1992. Meanwhile, the concentrations of ∑PAHs were positive correlated with total organic carbon contents except sites F and G, suggesting the characteristics of the sediment influences the distribution and concentration of PAHs. There was relatively a good relationship among the individual PAHs and the compounds of fluorene, phenanthrene, anthracene, fluoranthene, pyrene, benzo[a]pyrene and indeno[cd]pyrene yielded a good correlation (r(2) > 0.5) with total PAHs. Principal component analysis and specific PAHs compound ratios (Phe/Ant vs. Flt/Pyr) indicate the pyrogenic origins, especially traffic exhausts, are the dominant sources of PAHs in the Mai Po Inner Deep Bay Nature Reserve.

Disinfection characteristics of the dissolved organic fractions at several stages of a conventional drinking water treatment plant in Southern China.

Dissolved organic matter (DOM) influences many aspects of drinking water treatment, including the formation of harmful disinfection by-products (DBPs) when disinfectants are applied. DOM was isolated and fractionated using membrane ultra-filtration (UF) and reverse osmosis (RO) to eight individual fractions based on molecular weight cut-offs from a conventional surface water treatment plant (WTP) in Guangzhou of PR China. Molecular weights of these eight fractions were further calibrated using high performance size exclusion chromatography (HPSEC) and they ranged from 0.36 to 182.6 kDa. Fractions with molecular weight <0.80 kDa obtained by YC-05 UF membrane and RO were the major ones in all four stages of the water treatment processes; both ZM-500 and YM-100 membranes showed the highest removal efficiency when coupling with conventional coagulation and sedimentation processes. The elemental analysis showed that YC-05 fraction had greater polarity and aromaticity than any of the others. Furthermore, disinfection characteristics and trihalomethane formation potential (THMFP) were determined for all DOM fractions obtained in this study. YC-05 fraction was the major precursor for trihalomethane (THMs) formation among the samples tested and could be removed effectively by particulate activated carbon (PAC) adsorption. RO fraction could not be removed by PAC adsorption and, as a result, consumed more chlorine in the disinfection process. The results suggested that advanced drinking water treatment should focus on the removal of low molecular weight DOM in the source water.

Molecular size distribution of dissolved organic matter in water of the Pearl River and trihalomethane formation characteristics with chlorine and chlorine dioxide treatments.

River water sample was collected from Guangzhou section of the Pearl River to investigate soluble organic fractions and formation of trihalomethane (THMs) after chlorine and chlorine dioxide treatments. The water sample was passed through Amicon YC-05, YM-1, YM-3, YM-10, YM-30, YM-100 and ZM-500 series membranes after a pre-treatment. The molecular weight distribution and the specific ultra-violet absorbance (SUVA(254)) of each fraction obtained from membrane were analyzed, and these fractions were further disinfected with chlorine and chlorine dioxide. The results showed that reverse osmosis (RO) fraction contained mainly dissolved organic matter (DOM) from the water sample, suggesting that the water has been highly contaminated by anthropogenic activities. Meanwhile, the THMs concentration and SUVA(254) increased gradually as the molecular weight of the obtained fractions reduced, indicating that the low molecular weight DOM was the major THMs precursor in the disinfection process with chlorine and chlorine dioxide. The results suggest that THMs in source water of Pearl River could be effectively reduced when pollution of human activity is greatly controlled. Between the two disinfection processes tested, chlorine dioxide produced less THMs than chlorine in this study.

Chemical oxidative degradation of methyl tert-butyl ether in aqueous solution by Fenton's reagent.

Degradation of methyl tert-butyl ether (MTBE) in aqueous solution by Fenton's reagent (Fe2+ and H2O2) was investigated. Effects of reaction conditions on the oxidation efficiency of MTBE by Fenton's reagent were examined in batch experiments. Under optimum conditions, 15 mM H2O2, 2 mM Fe2+, pH 2.8 and room temperature, the initial 1 mM MTBE solution was reduced by 99% within 120 min. Results showed that MTBE was decomposed in a two-stage reaction. MTBE was first decomposed swiftly based on a Fe2+/H2O2 reaction and then decomposed somewhat less rapidly based on a Fe3+/H2O2 reaction. The detection of Fe2+ also supported the theory of the two-stage reaction for the oxidation of MTBE by Fenton's reagent. The dissolved oxygen in the solution decreased rapidly in the first stage reaction, but it showed a slow increase in the second stage with a zero-order kinetics. A reaction mechanism involving two different pathways for the decomposition of MTBE by Fenton's reagent was also proposed. Chemicals including tert-butyl formate, tert-butyl alcohol, methyl acetate and acetone were identified to be the primary intermediates and by-products of the degradation processes.

[Effect on trihalomethane forming for different molecular weight dissolved organic matter in water resource].

A raw water sample was collected from Guangzhou section of Pearl River to support experiment studies. The sample were passed through Amicon YC-05, YM-1, YM-3, YM-10, YM-30, YM-100 and ZM-500 series membranes after being pretreated. The molecular weight distribution of the isolate for each membranes were analyzed, and the isolates were disinfected by chlorine and chlorine dioxide. The results showed that the RO isolate was the main dissolved organic matter (DOM) in the water sample, which proved that the water was highly contaminated by human activities, and it was the main THMs precursor in the disinfection process with chlorine and chlorine dioxide for all the isolates. Meanwhile, the disinfection process for water sample contain DOM that its molecular smaller than 500 were very different from raw water.