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The spread of methicillin-resistant Staphylococcus aureus (MRSA) poses a major threat to global health. Isothermal amplification combined with the trans -cleavage activity of Cas12a has been exploited to generate diagnostic platforms for pathogen detection.

Purpose: Due to poor prognosis and immunotherapy failure of skin cutaneous melanoma (SKCM), this study sought to find necroptosis-related biomarkers to predict prognosis and improve the situation with predicted immunotherapy drugs. Experimental Design: The Cancer Genome Atlas (TCGA) and The Genotype-Tissue Expression Program (GTEx) database were utilized to recognize the differential necroptosis-related genes (NRGs). Univariate Cox (uni-Cox) and least absolute shrinkage and selection operator (LASSO) Cox analysis were utilized for prognostic signature establishment. The signature was verified in the internal cohort. To assess the signature’s prediction performance, the area under the curve (AUC) of receiver operating characteristic (ROC) curves, Kaplan-Meier (K-M) analyses, multivariate Cox (multi-Cox) regression, nomogram, and calibration curves were performed. The molecular and immunological aspects were also reviewed using single-sample gene set enrichment analysis (ssGSEA). Cluster analysis was performed to identify the different types of SKCM. Finally, the expression of the signature gene was verified by immunohistochemical staining. Results: On basis of the 67 NRGs, 4 necroptosis-related genes (FASLG, PLK1, EGFR, and TNFRSF21) were constructed to predict SKCM prognosis. The area’s 1-, 3-, and 5-year OS under the AUC curve was 0.673, 0.649, and 0.677, respectively. High-risk individuals had significantly lower overall survival (OS) compared to low-risk patients. Immunological status and tumor cell infiltration in high-risk groups were significantly lower, indicating an immune system that was suppressed. In addition, hot and cold tumors could be obtained by cluster analysis, which is helpful for accurate treatment. Cluster 1 was considered a hot tumor and more susceptible to immunotherapy. Immunohistochemical results were consistent with positive and negative regulation of coefficients in signature. Conclusion: The results of this finding supported that NRGs could predict prognosis and help make a distinction between the cold and hot tumors for improving personalized therapy for SKCM.

We constructed the first high-quality genome of E. cyanopodus based on DNBSEQ-T1, PacBio, and Hic sequencing technologies, with a genome size of 998.82 Mb, contig N50 of 5.855 Mb.

The water-droplet erosion of low-pressure steam turbine blades under wet steam environments can alter the vibration characteristics of the blade, and lead to its premature failure. Using high-velocity oxygen-fuel (HVOF) sprayed water-droplet erosion resistant coating is beneficial in preventing the erosion failure, while the erosion behavior of such coatings is still not revealed so far. Here, we examined the water-droplet erosion resistance of Cr3C2–25NiCr and WC–10Co–4Cr HVOF sprayed coatings using a pulsed water jet device with different impingement angles. Combined with microscopic characterization, indentation, and adhesion tests, we found that: (1) both of the coatings exhibited a similar three-stage erosion behavior, from the formation of discrete erosion surface cavities and continuous grooves to the broadening and deepening of the groove, (2) the erosion rate accelerates with the increasing impingement angle of the water jet; besides, the impingement angle had a nonlinear effect on the cumulative mass loss, and 30° sample exhibited the smallest mass loss per unit area (3) an improvement in the interfacial adhesion strength, fracture toughness, and hardness of the coating enhanced the water-droplet erosion resistance. These results provide guidance pertaining to the engineering application of water erosion protective coatings on steam turbine blades.

Protein kinases play crucial roles in the regulation of plant resistance to various stresses. In this work, we determined that GsSnRK1.1 was actively responsive to saline-alkali, drought, and abscisic acid (ABA) stresses by histochemical staining and qRT-PCR analyses. The wild-type GsSnRK1.1 but not the kinase-dead mutant, GsSnRK1.1(K49M), demonstrated in vitro kinase activity by phosphorylating GsABF2. Intriguingly, we found that GsSnRK1.1 could complement the loss of SNF1 kinase in yeast Msy1193 (-snf1) mutant, rescue growth defects of yeast cells on medium with glycerol as a carbon resource, and promote yeast resistance to NaCl or NaHCO

Paclitaxel (Taxol), a highly modified diterpene agent mainly obtained from

Chrysanthemum is one of the most important ornamental flowers in the world. In general, abnormal morphology of Chrysanthemum flowers is developed during~35–40 °C, which significantly damages incomes of flower gardener. Therefore, to select Chrysanthemum lines of high temperature tolerance is of vital importance. In this study, transgenic calluses with eIF5B1, a regulatory gene of high temperature tolerance, were generated, and the presence of the transgene and its stable expression were confirmed through PCR, GUS assay and Western blotting. These transgenic calluses with plantlets were also observed by both electron and optical microscope. Findings indicate these lines contain more starch granules, which are a marker of high temperature tolerance. We expect that the efficient protocol developed in this study will facilitate the genetic transformation of the genes of interest and that these regulatory genes will help in keeping the flower morphology of Chrysanthemum under high temperature stress.

Background Allium sativum (garlic) is an economically important food source and medicinal plant rich in sulfides and other protective substances such as alliin, the precursor of allicin biosynthesis. Cysteine, serine and sulfur is the precursor of alliin biosynthesis. However, little is known about the alliin content under abiotic stress or the mechanism by which it is synthesized. Results The findings revealed that the content of alliin was lowest in the garlic roots, and highest in the buds. Furthermore, alliin levels decreased in mature leaves following wounding. Transcriptome data generated over time after wounding further revealed significant up-regulation of genes integral to the biosynthetic pathways of cysteine and serine in mature garlic leaves. Conclusions The findings suggest that differential expression of cysteine, serine and sulfide-related genes underlies the accumulation of alliin and its precursors in garlic, providing a basis for further analyses of alliin biosynthesis.

Polarimetric LiDAR can suppress backscattering by aerosols and increase detection contrast. Depolarization by the observed object can hinder those efforts. We present linear and circular depolarization ratios from 61 targets and the impact on contrast.

Additional file 3: Figure S2. Volcano plot of T1/T3 vs. T7/T9, T1/T3 vs. T7/9 and T1/T3 vs. T10/12 of upgenes and downgenes.

Additional file 2: Figure S1. Differential expression of KEGG pathway-related genes.

Additional file 4: Figure S3. The figure is unigenes contrasted to the COG database.

Cytosolic invertase 1 (CINV1), which converts sucrose into glucose and fructose is a key entry point of carbon into cellar metabolism and Hexokinase 1 (HXK1) functions as a pivotal sensor for glucose. Further, it is well established that sugar signaling can regulate plant gene expression, for example, the Production of Anthocyanin Pigment 1 (PAP1) / MYB75 transcription factor is specifically induced by sucrose accumulation. Exogenous sugars are also thought to trigger the juvenile-to-adult phase transition via a miR156A/SQUAMOSA PROMOTER BINDING PROTEIN-LIKE (SPL) module. However, the endogenous factors that regulate this process have remained undetermined. Here we show that sucrose induced expression of the PAP1 transcription factor increases CINV1 activity by direct binding to CINV1 promoter. Further, we identify a glucose feed-forward loop (Sucrose-CINV1-Glucose-HXK1-miR156-SPL9-PAP1-CINV1-Glucose) that controls CINV1 activity to convert sucrose into glucose signaling to dynamically control the juvenile-to-adult phase transition. Moreover, PAP1 directly binds to the SPL9 promoter, activating SPL9 expression, triggering sucrose signaling that mediates the juvenile-to-adult phase transition. Therefore, a glucose signaling feed-forward loop and a sucrose signaling pathway synergistically regulate the Arabidopsis juvenile-to-adult phase transition. Collectively, we identify a molecular link between the major photosynthate sucrose, the entry point of carbon into cellar metabolism and a key developmental process, the juvenile-to-adult phase transition.

Most of the closely related fish species are similar in appearance, with slight differences. This study evaluated 35 morphological characteristic parameters and the number, shape, and spatial location of bones in the skull and vertebrae of the mandarin fish Siniperca chuatsi and big-eye mandarin fish Siniperca kneri. We can understand their ecological adaptability, compare their morphological and skeletal differences, and elucidate the relationship between their phenotypic trait differences and skeletal characteristics. The results were as follows: 1. The Student's t-test results revealed significant differences between S. chuatsi and S. kneri in terms of head depth, head back length, snout length, interorbital distance, distance of the eyes from the oral fissure, and width between the posterior trunk and base of the caudal fin. Principal component and discriminant analysis showed that the main morphological differences between S. chuatsi and S. kneri were in the eye position, head hump and body shape. 2. There was no significant difference in the number and shape of the skull and vertebrae between S. chuatsi and S. kneri, but they differed in spatial position. The terminus of the maxilla in S. chuatsi is located at the lower border of the postorbital bone, but at the anterior in S. kneri. The size of head hump is closely related to the spatial location of the splanchnocranium in S. chuatsi and S. kneri. The angles between the caudal vertebrae centrum and each of the following: the neural arch and spine; haemal arch and spine of S. chuatsi are larger than those of S. kneri. In conclusion, the morphological differences between S. chuatsi and S. kneri are closely related to the spatial position of skeleton. These phenotypic differences reflect the ecological adaptation of mandarin fish to different natural environments.

Anthocyanin accumulation specifically depends on sucrose (Suc) signaling. However, the molecular basis of this process remains unknown. In this study, in vitro pull-down assays identified ETHYLENE-INSENSITIVE3 (EIN3), a component of both sugar signaling or/and metabolism. This protein interacted with YDA, and the physiological relevance of this interaction was confirmed by in planta co-immunoprecipitation, yeast two-hybrid (Y2H) assay, and bimolecular fluorescence complementation. Ethylene insensitive3-like 1 (eil1) ein3 double-mutant seedlings, but not ein3-1 seedlings, showed anthocyanin accumulation. Furthermore, ein3-1 suppressed anthocyanin accumulation in yda-1 plants. Thus, EMB71/YDA-EIN3-EIL1 may form a sugar-mediated gene cascade integral to the regulation of anthocyanin accumulation. Moreover, the EMB71/YDA-EIN3-EIL1 gene cascade module directly targeted the promoter of Transparent Testa 8 (TT8) by direct EIN3 binding. Collectively, our data inferred a molecular model where the signaling cascade of the YDA-EIN3-TT8 appeared to target TT8 via EIN3, thereby modulating Suc signaling–mediated anthocyanin accumulation.

Light signals are perceived by multiple photoreceptors that converge to suppress the RING E3 ubiquitin ligase CONSTITUTIVE PHOTOMORPHOGENIC1 (COP1) for the regulation of stomatal development. Thus, COP1 is a point of integration between light signaling and stomatal patterning. However, how light signaling is collected into COP1 for the production and spacing of stomata is still unknown. Here, we report that the loss-of-function mutant of ANGUSTIFOLIA3 (AN3) delays asymmetric cell division, which leads to decreased stomatal index. Furthermore, overexpression of AN3 accelerates asymmetric cell division, which results in clusters of stomata. In addition, the stomatal development through AN3 regulation is mediated by light signaling. Finally, we find that an3 is a light-signaling mutant, and that AN3 protein is light regulated. Self-activation by AN3 contributes to the control of AN3 expression. Thus, AN3 is a point of collection between light signaling and stomatal patterning. Target-gene analysis indicates that AN3 is associated with COP1 promoter for the regulation of light-controlling stomatal development. Together, these components for regulating stomatal development form an AN3-COP1-E3 ubiquitin ligase complex, allowing the integration of light signaling into the production and spacing of stomata.

Euphorbia pekinensis Radixz (EPR) is a commonly used Chinese herbal medicine prepared from the dried roots of E. pekinensis Rupr. It has been revealed in chemical analysis that diterpenoids are the primary component of EPR bestowing toxicity and bioactivity; however, little is known about these bioactive and toxic diterpenoid biosynthesis pathways and diterpene diversity in E. Pekinensis. In this study, we sequenced root, stem, and leaf transcriptomes of E. pekinensis, and conducted de novo assembly using 429 million clean reads with Trinity software. Then, 157,491 unigenes were generated whose N50 contained 1826 bp. All of the unigenes were interpreted using Nt, Nr, COGs, SwissProt, GO, and KEGG databases. The expression profiling based on FPKM showed 665 genes up-regulated in roots were enrichment pathways for CYP450s and transcript factors. There are extremely diverse specialised diterpene pathways therein, and most specialised diterpenes are limited to several plant genera, species, or special tissues according to their taxonomy. Using homology, we identified 26 diterpenoid synthase candidates which comprised 25 class I proteins, and only one class II protein, in TPS-a and TPS-c sub-families, among which five class I proteins were highly expressed in root. Additionally, 23 CYP450 enzymes from sub-families of CYP71D and CYP726 were identified, and three were highly expressed in root, for which expression patterns were similar to the aforementioned five class I diTPSs expressed in root and may be involved in macroditerpenoid biosynthesis in EPR. Our results provided a beneficial genomic resource which is able to be used for future functional genomics research on E. pekinensis.

As a hermaphroditic fish with considerable economic value, the Hong Kong grouper (Epinephelus akaara) has suffered a continuous decline in wild resources from long-term overfishing and habitat destruction, while with artificial propagation, the low proportion of male fish has always been a difficult problem to address. Understanding the gonadal structure and sex differentiation mechanism of this fish is the key to proposing effective solutions. For many hermaphroditic species, the gonad is observed to be separated into testicular and ovarian parts by a physical boundary. Notably, in this study, no such structure was found in Hong Kong grouper, implying the uniqueness of its gonadal structure, which may be associated with plastic sex. The analysis of important sex-related genes in Hong Kong grouper showed that the expression pattern of vasa, dmrt1, and foxl2 was similar to that of most gonochoristic and other hermaphroditic fishes, indicating the conserved functions of those genes. Unlike previous descriptions in other fish species, the unique expression pattern of foxl3 in Hong Kong grouper revealed its potential function in both female- and male-phase gonads, and we suspected that it might play a crucial role in testicular tissue development and maintenance of functions. This study enables us to obtain a better understanding of sex differentiation in Hong Kong grouper and provides theoretical guidance for artificial breeding in aquaculture.

The formation and the development of the lateral organs require the expression of organ-specific genes. On the other hand, meristem-specific genes are repressed to complete the differentiation of the organs. KNOTTED-like home-obox (KNOX) genes have been isolated from several plant species. Genetic analysis has demonstrated the role of KNOX class I genes in the formation and maintenance of the shoot apical meristems (SAMs). The ASYMMETRIC LEAVES2 (AS2)/LATERAL ORGAN BOUNDARIES (LOB) gene of Arabidopsis defines a novel conserved protein domain. LOB, the founding member of this gene family, is considered as playing a role in boundary establishment or communication between the meristems and initiating lateral organs. With organ initiation, roles of LBD genes (such as AS2, ASL1) cause that the expression of KNOX genes is downregulated in later organ, this downregulation is critical for the leaf primordial development and subsequently proper explanation of leaf blades. However,JLO/LBD30 misexpression induces ectopic expression of KNOX genes, i.e., STM and KNAT1, and overexpression of ASL11/LBD15 and ASL38/LBD41 cockscomb plants causes lob leaves and pin-like-SAMs, suggesting LBD genes play a novel role in lateral organ development. A possibility is JLO/LBD30 as an LBD transcription factor can coordinate two processes (Auxin accumulation and loss of meristem-specific gene expression in organ primordial) by controlling both auxin transport and the expression of KNOX genes. Thus, this review mainly addresses how the antagonistic or synchronous relationship between ASL/LBD and homeobox members forms complex signaling networks.

ASYMMETRIC LEAVES2-LIKE16 gene of Arabidopsis is a member of the ASYMMETRIC LEAVES2 (AS2)/LATERAL ORGAN BOUNDARIES (LOB) domain gene family. Current reports have indicated that overexpression of ASL16 induces lateral root formation in the absence of ARF7 and ARF19; whereas lateral root formation is severely reduced in the arf 7 and arf 19 double knockout mutants. Antisense mRNA has testified to be a useful tool to inhibit the expression of specific genes in plants; more significantly, it is specific in the sense that only the mRNA level of the desired protein being changed, while other members of the gene family remain unaffected. To further explore ASL16 function in lateral root growth and development, we introduced an antisense-expression construct of the ASL16 gene to Arabidopsis (Col-0); and acquired some 35S:ASL16 Arabidopsis plants. With 11 day after germination (DAG), many lateral roots in wild-type seedlings were well developed, whereas a few lateral roots in these antisense 35S:ASL16 seedlings were produced. However, with 5, 8 and 16 DAG, the lateral roots in these transgenic seedlings were not distinct with those of wild-type plants. A series of data indicate that primary root formation of these antisense 35S:ASL16 plants was not affected; whereas their lateral root formation was severely inhibited.

A vertically scanning lidar (light/radar) was used to measure the time evolution of clouds generated by a small explosive device. Vertical sweeps were performed at a downwind distance of 105 m from the detonation. The measured quantity obtained from the lidar was the light extinction coefficient. This quantity is directly proportional to the aerosol concentration. The background aerosol value was set to 0.0001 m (-1) (assuming a visibility of 40 km), and assuming the scattering properties of the explosively generated cloud is the same as the background aerosol, the authors found that the instantaneous maximal local concentration of aerosol in the cloud did not exceed 500 times the background aerosol value, and the instantaneous concentration was typically less than five times the background aerosol value. In the two trials that were done, the volumes of the clouds were reasonably close at 2,700 m(3) and 4,000 m(3), respectively.

Lane changing is one of the basic driver behavior interactions in microscopic traffic simulations for traffic, safety, and transportation system analysis. Although many traffic simulations pay attention to the decision process for lane changing, the execution process is often simplified or even ignored. This paper presents a study of lane change execution of heavy vehicles and passenger cars in a traffic situation with a blocked lane. A video camera was used to collect data from an arterial road in Melbourne, Australia. This paper investigates the behavior of heavy vehicles and passenger cars in mandatory lane changes and proposes a model framework for the execution of lane changes with regard to the emergency status and the impact of surrounding traffic for individual drivers who are changing lanes. The distance to the blockage was used to denote the emergency status. For the impact of the surrounding traffic, it was assumed that the driver would adjust the execution if a traffic conflict were detected and would continue the lane change if there were no conflict. A probability model was developed to interpret the driver’s choice of whether to continue the lane change. The data analysis found that the lane change performance of heavy vehicles differed from that of passenger cars. Therefore, the model was calibrated for heavy vehicles and passenger cars separately. In conclusion, the paper provides a framework of the future work of lane change execution models on traffic simulation to assess the traffic safety and road efficiency.

Squalene synthase catalyzes the condensation of 2 molecules of farnesyl diphosphate to produce squalene, the first committed precursor for sterol, brassinosteroid, and triterpene biosynthesis. A squalene synthase gene, designated IoSQS, was isolated from Inonotus obliquus, a medicinal mushroom that produces a plethora of bioactive triterpenes. IoSQS complementary DNA was found to contain an open reading frame of 1476 bp, encoding a protein of 491 amino acids with a calculated molecular mass of 55.85 kDa. The IoSQS genomic DNA sequence consisted of 1813 bp and contained 4 exons and 3 introns. The restriction fragment polymorphisms revealed by Southern blot analysis suggested that IoSQS was a single-copy gene. Promoter analysis indicated that the 5' upstream region of IoSQS possessed various potential elements associated with physiological and environmental factors. The expression pattern of IoSQS in different stages and under methyl jasmonate treatment correlated with the accumulation of total triterpenoids and was consistent with the predicted results of the IoSQS promoter region. The N-terminal 466 residues of the hydrophilic sequence were expressed as a His-tagged protein in Escherichia coli, and the resultant bacterial crude extract was incubated with farnesyl diphosphate and NADPH. Squalene was detected in vitro in reaction mixture by high-performance liquid chromatography analysis. These results suggest that the IoSQS enzyme is involved in squalene production in I. obliquus.

Snow precipitation interaction with a generic 3D lidar is modeled. The randomness and the intensity of the signal as a function of the visibility and snowflake size and density distribution are reproduced. To do so, a representative snow density distribution is modeled as a function of visibility. Taking into account the laser beam and pulse characteristics, the probability to have one or many snowflakes of a given size in the lidar sampling cell is calculated. Knowing the number and the size of the snowflakes, the magnitude of the lidar signal is calculated. Finally, a filtering algorithm based on the relative intensity of the snowflakes is discussed.

The backscattering lidar depolarization parameter D of water droplets contains information on their size that can be directly modeled as a function of the forward-scattering diffraction peak. Using a polarimetric Monte Carlo simulator, water clouds having different extinctions and droplet size distributions are analyzed to estimate their depolarization parameter at various backscattering off-axis angles. It is shown that the depolarization parameter of the polarimetric phase function can be found using off-axis lidar measurements at multiple angles, and that it could be used to estimate the water-cloud droplet size.

The contrast in the azimuthal pattern of cross-polarized lidar data is used directly to retrieve the extinction coefficient profile of water droplet clouds. Using a Monte Carlo simulation, we demonstrate that there is a simple mathematical relationship between the optical depth and the contrast of the cross-polarization azimuthal pattern. This relationship is independent of the water cloud droplet size, cloud position, and extinction profile. The derivation of the extinction profile of a water droplet cloud is obtained directly using the simple mathematical relationship without performing lidar equation inversion. The technique is limited to spherical particles.

BACKGROUND Heart sound monitor (HSM), a device suitable for home-use, can be used to record heart sounds and to store, transmit or analyze those recordings. It enables cardiac reserve telemonitoring which has been largely evolved and widely used in recent years. Nevertheless, the designers of HSM paid little attention to the consistency of its information model and data interaction, and because of that, the collected heart sounds data could not be shared and aggregated effectively. Thus, the device’s development and its application in telehealth service are hindered. OBJECTIVE In order to solve the above problem and to build interoperability for HSM devices, this paper proposes a HSM interoperability framework that is composed of the hierarchical information model and the transport-independent interaction model, which is constructed by using standardized modeling methods. METHODS The authors collected and studied the common device-output information of HSM involved in the cardiac reserve telemonitoring, leveraged the standardized interoperability framework defined in ISO/IEEE 11073 Personal Health Device (11073-PHD) standards to model the static data structure and dynamic interaction behaviors of HSM. RESULTS Via the meta analysis, the authors identified that the common device-output information of HSM mainly includes the phonocardiogram (PCG), the feature parameters of PCG, the ratio of diastolic to systolic duration (D/S), for example, and the threshold data, device status, sensor location, etc. Based on such information, an 11073-PHD-compliant domain information model has been successfully created. This enables the interoperability between HSM and aggregation device, allowing inter-device plug&play using service model and communication model. A prototype of this design has been implemented and validated via the Continua Enabling Software Library. CONCLUSIONS The ISO/IEEE 11073-PHD standard framework has the potential to accommodate the HSM device. The standard-compliant domain information models can be established to cover the common HSM device-output information. Findings in this paper may be taken as a reference for standard developing organizations to establish a standardized interoperability framework for HSM.

This thesis explores lane change execution behaviour in heterogeneous traffic, with a focus on mandatory lane changes on arterial roads. Existing lane change studies mostly pay attention to the lane change generation process, while the lane change execution process is often simplified or even ignored. The lane changing of different vehicle types is also not investigated satisfactorily. To address these knowledge gaps, this research has improved the understanding of mandatory lane change execution, identified the influencing factors to lane change execution and furthermore developed lane change execution models for both passenger cars and heavy vehicles.

Information about the size distribution of liquid droplets in a fog can be retrieved by measuring the backscattering lidar depolarization parameter D in circular polarization. Using a polarimetric off-axis lidar, measurements at different backscattering angles are performed on fogs made of water droplets and of mineral oil. Estimation of the effective droplet size is obtained using constrained linear inversion. Mie theory is used to calculate the variation in depolarization parameters for different effective droplet sizes. The calculation is performed for various scattering angles. These calculations provide a kernel for the constrained linear inversion scheme. It is shown that the refractive index has an effect on the retrieved droplet sizes as well as the choice of scattering angles. These measurements confirm that the circular depolarization parameter measured near the backscattering angle can be modeled as a function of the forward-scattering diffraction peak. The results of the constrained linear inversion of measurements are consistent with in situ measurement of the droplet size distribution.