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Abstract Background Most tail-anchored (TA) membrane proteins are delivered to the endoplasmic reticulum through a conserved posttranslational pathway. Although core mechanisms underlying the targeting and insertion of TA proteins are well established in eukaryotes, their role in mediating TA protein biogenesis in plants remains unclear. We reported the crystal structures of algal arsenite transporter 1 (ArsA1), which possesses an approximately 80-kDa monomeric architecture and carries chloroplast-localized TA proteins. However, the mechanistic basis of ArsA2, a Get3 (guided entry of TA proteins 3) homolog in plants, for TA recognition remains unknown. Results Here, for the first time, we present the crystal structures of the diatom Pt-Get3a that forms a distinct ellipsoid-shaped tetramer in the open (nucleotide-bound) state through crystal packing. Pulldown assay results revealed that only tetrameric Pt-Get3a can bind to TA proteins. The lack of the conserved zinc-coordination CXXC motif in Pt-Get3a potentially leads to the spontaneous formation of a distinct parallelogram-shaped dimeric conformation in solution, suggesting a new dimer state for subsequent tetramerization upon TA targeting. Pt-Get3a nonspecifically binds to different subsets of TA substrates due to the lower hydrophobicity of its α-helical subdomain, which is implicated in TA recognition. Conclusions Our study provides new insights into the mechanisms underlying TA protein shielding by tetrameric Get3 during targeting to the diatom’s cell membrane.

Microwave radiometry has provided valuable spaceborne observations of Earth's geophysical properties for decades. The recent SMOS, Aquarius, and SMAP satellites have demonstrated the value of measurements at 1400 MHz for observing surface soil moisture, sea surface salinity, sea ice thickness, soil freeze/thaw state, and other geophysical variables. However, the information obtained is limited by penetration through the subsurface at 1400 MHz and by a reduced sensitivity to surface salinity in cold or wind-roughened waters. Recent airborne experiments have shown the potential of brightness temperature measurements from 500–1400 MHz to address these limitations by enabling sensing of soil moisture and sea ice thickness to greater depths, sensing of temperature deep within ice sheets, improved sensing of sea salinity in cold waters, and enhanced sensitivity to soil moisture under vegetation canopies. However, the absence of significant spectrum reserved for passive microwave measurements in the 500–1400 MHz band requires both an opportunistic sensing strategy and systems for reducing the impact of radio-frequency interference. Here, we summarize the potential advantages and applications of 500–1400 MHz microwave radiometry for Earth observation and review recent experiments and demonstrations of these concepts. We also describe the remaining questions and challenges to be addressed in advancing to future spaceborne operation of this technology along with recommendations for future research activities.

The Cubesat radiometer radio frequency interference technology validation mission (CubeRRT) was developed to demonstrate real-time onboard detection and filtering of radio frequency interference (RFI) for wide bandwidth microwave radiometers. CubeRRT's key technology is its radiometer digital backend (RDB) that is capable of measuring an instantaneous bandwidth of 1 GHz and of filtering the input signal into an estimated total power with and without RFI contributions. CubeRRT's onboard RFI processing capability dramatically reduces the volume of data that must be downlinked to the ground and eliminates the need for ground-based RFI processing. RFI detection is performed by resolving the input bandwidth into 128 frequency subchannels, with the kurtosis of each subchannel and the variations in power across frequency used to detect nonthermal contributions. RFI filtering is performed by removing corrupted frequency subchannels prior to the computation of the total channel power. The 1 GHz bandwidth input signals processed by the RDB are obtained from the payload's antenna (ANT) and radiometer front end (RFE) subsystems that are capable of tuning across RF center frequencies from 6 to 40 GHz. The CubeRRT payload was installed into a 6U spacecraft bus provided by Blue Canyon Technologies that provides spacecraft power, communications, data management, and navigation functions. The design, development, integration and test, and on-orbit operations of CubeRRT are described in this article. The spacecraft was delivered on March 22nd, 2018 for launch to the International Space Station (ISS) on May 21st, 2018. Since its deployment from the ISS on July 13th, 2018, the CubeRRT RDB has completed more than 5000 h of operation successfully, validating its robustness as an RFI processor. Although CubeRRT's RFE subsystem ceased operating on September 8th, 2018, causing the RDB input thereafter to consist only of internally generated noise, CubeRRT's key RDB technology continues to operate without issue and has demonstrated its capabilities as a valuable subsystem for future radiometry missions.

Purpose - To support the standardized evaluation of bicyclist automatic emergency braking (AEB) systems, test scenarios, test procedures and test system hardware and software tools have been investigated and developed by the Transportation Active Safety Institute (TASI) at Indiana University-Purdue University Indianapolis. This paper aims to focus on the development of test scenarios and bicyclist surrogate for evaluating vehicle–bicyclist AEB systems. Design/methodology/approach - The harmonized general estimates system (GES)/FARS 2010-2011 crash data and TASI 110-car naturalistic driving data (NDD) are used to determine the crash geometries and environmental factors of crash scenarios including lighting conditions, vehicle speeds, bicyclist speeds, etc. A surrogate bicyclist including a bicycle rider and a bicycle surrogate is designed to match the visual and radar characteristics of bicyclists in the USA. A bicycle target is designed with both leg pedaling and wheel rotation to produce proper micro-Doppler features and generate realistic motion for camera-based AEB systems. Findings - Based on the analysis of the harmonized GES/FARS crash data, five crash scenarios are recommended for performance testing of bicyclist AEB systems. Combined with TASI 110-car naturalistic driving data, the crash environmental factors including lighting conditions, obscuring objects, vehicle speed and bicyclist speed are determined. The surrogate bicyclist was designed to represent the visual and radar characteristics of the real bicyclists in the USA. The height of the bicycle rider mannequin is 173 cm, representing the weighted height of 50th percentile US male and female adults. The size and shape of the surrogate bicycle were determined as 26-inch wheel and mountain/road bicycle frame, respectively. Both leg pedaling motion and wheel rotation are suggested to produce proper micro-Doppler features and support the camera-based AEB systems. Originality/value - The results have demonstrated that the developed scenarios, test procedures and bicyclist surrogate will provide effective objective methods and necessary hardware and software tools for the evaluation and validation of bicyclist AEB systems. This is crucial for the development of advanced driver assistance systems.

Thraustochytrids are heterotrophic fungus-like protists that can dissolve organic matters with enzymes. Four strains, AP45, ASP1, ASP2, and ASP4, were isolated from the coastal water of Taiwan, and respectively identified as Aurantiochytrium sp., Schizochytrium sp., Parietichytrium sp., and Botryochytrium sp. based on 18S rRNA sequences. Transcriptome datasets of these four strains at days 3-5 were generated using Next Generation Sequencing technology, and screened for enzymes with potential industrial applications. Functional annotations based on KEGG database suggest that many unigenes of all four strains were related to the pathways of industrial enzymes. Most of all four strains contained homologous genes for 15 out of the 17 targeted enzymes, and had extra- and/or intra-cellular enzymatic activities, including urease, asparaginase, lipase, glucosidase, alkaline phosphatase and protease. Complete amino sequences of the first-time identified L-asparaginase and phytase in thraustochytrids were retrieved, and respectively categorized to the Type I and BPPhy families based on phylogenetic relationships, protein structural modeling and active sites. Milligram quantities of highly purified, soluble protein of urease and L-asparaginase were successfully harvested and analyzed for recombinant enzymatic activities. These analytical results highlight the diverse enzymes for wide-range applications in thraustochytrids.

The feedback strategy, or controlled exposure of pig herd to the porcine epidemic diarrhea virus (PEDV), significantly decreased losses during a severe outbreak in late 2013 in Taiwan. However, some pig farms still suffered from recurrent outbreaks. To evaluate the association between antibody titers and clinical manifestations, sera and colostra were analyzed from one pig farm that employed the feedback strategy. Furthermore, spike (S) gene full sequences from six positive samples of two farms with and without using feedback were compared to investigate the evolution of PEDV variants circulating in pig herds. The results in this study showed that high PEDV antibody titers do not correlate with the high rate of protection from PEDV infection. In addition, repeated feedback generated the emergence of PEDV variants with unique substitutions of N537S and Y561H in the COE domain and S769F in the SS6 epitopes. These mutations indicated the pathogenetic evolution of PEDV strains existing in the cycle of the feedback method. A very strict biosecurity practice to block the routes of pathogen transfer should be followed to achieve successful control of PEDV infections in pig herds.

Cooling the trapped atoms toward their motional ground states is key to applications of quantum simulation and quantum computation. By utilizing nonreciprocal couplings between two atoms, we present an intriguing dark-state cooling scheme in Λ-type three-level structure, which is shown superior than the conventional electromagnetically-induced-transparency cooling in a single atom. The effective nonreciprocal couplings can be facilitated either by an atom–waveguide interface or a free-space photonic quantum link. By tailoring system parameters allowed in dark-state cooling, we identify the parameter regions of better cooling performance with an enhanced cooling rate. We further demonstrate a mapping to the dark-state sideband cooling under asymmetric laser driving fields, which shows a distinct heat transfer and promises an outperforming dark-state sideband cooling assisted by collective spin–exchange interactions.

Dilute ethanol (EtOH) is a widely used agent to remove the corneal epithelium during the modern refractive surgery. The application of EtOH may cause the underlying corneal fibroblasts to undergo apoptosis. This study was designed to investigate the protective effect and potential mechanism of the respiratory chain coenzyme Q(10) (CoQ(10)), an electron transporter of the mitochondrial respiratory chain and a ubiquitous free radical scavenger, against EtOH-induced apoptosis of corneal fibroblasts. Corneal fibroblasts were pretreated with CoQ(10) (10 µM) for 2 h, followed by exposure to different concentrations of EtOH (0.4, 2, 4, and 20%) for 20 s. After indicated incubation period (2-12 h), MTT assay was used to examine cell viability. Treated cells were further assessed by flow cytometry to identify apoptosis. Reactive oxygen species (ROS) and the change in mitochondrial membrane potential were assessed using dichlorodihydrofluorescein diacetate/2',7'-dichlorofluorescein (DCFH-DA/DCF) assays and flow-cytometric analysis of JC-1 staining, respectively. The activity and expression of caspases 2, 3, 8, and 9 were evaluated with a colorimetric assay and western blot analysis. We found that EtOH treatment significantly decreased the viability of corneal fibroblasts characterized by a higher percentage of apoptotic cells. CoQ(10) could antagonize the apoptosis inducing effect of EtOH. The inhibition of cell apoptosis by CoQ(10) was significant at 8 and 12 h after EtOH exposure. In EtOH-exposed corneal fibroblasts, CoQ(10) pretreatment significantly reduced mitochondrial depolarization and ROS production at 30, 60, 90, and 120 min and inhibited the activation and expression of caspases 2 and 3 at 2 h after EtOH exposure. In summary, pretreatment with CoQ(10) can inhibit mitochondrial depolarization, caspase activation, and cell apoptosis. These findings support the proposition that CoQ(10) plays an antiapoptotic role in corneal fibroblasts after ethanol exposure.