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The glycolytic enzyme cytoplasmic glyceraldehyde-3-phosphate dehydrogenase (GAPC3) is involved in multiple biological processes in plants, including transcriptional regulation, and material metabolism. However, the relationship between OsGAPC3 and the quality traits of rice is poorly understood. Here we identify OsGAPC3 mutations that enhance the protein content and grain nutritional quality of rice by regulating the OsAAP6 gene expression. The number and volume of type-II protein bodies in the endosperm of the OsGAPC3 mutants, and GPC increase significantly. We report significant increases in chalkiness area and degree, and decreases for starch content, gel consistency, and taste value. Results of proteomic detection and analysis reveal that OsGAPC3 affects the major storage substances (proteins and starch) metabolism in rice, and the accumulation of proteins and starch in the endosperm. Additionally, the OsGAPC3 mutation significantly decreases the rice-seedling salt tolerance. Therefore, OsGAPC3 affects multiple quality traits of rice, participates in regulating rice-seedling salt-stress response. These data can be used to design better-quality and stronger salt-resistant rice varieties.

Abstract Bacillus cereus is a clinically significant foodborne pathogen that causes severe gastrointestinal and non-gastrointestinal disease. Cereolysin O (CLO) is a putative virulence factor of B. cereus, and its function remains to be investigated. In this study, we examined the biological activity of CLO from a deep sea B. cereus isolate. CLO was highly toxic to mammalian cells and triggered pyroptosis through NLRP3 inflammasome-mediated caspase 1 and gasdermin D activation. CLO-induced cell death involved ROS accumulation and K+ efflux, and was blocked by serum lipids. CLO bound specifically to cholesterol, and this binding was essential to CLO cytotoxicity. The structural integrity of the three tryptophan residues in the C-terminal undecapeptide was vital for CLO to interact with membrane lipids and cause membrane perforation. Taken together, these results provided new insights into the molecular mechanism of B. cereus CLO-mediated cytotoxicity.

Dynamic α precipitation with hot deformation is commonly adopted to obtain equiaxed α for tailoring the mechanical properties of Ti alloys. The formation of an equiaxed shape rather than the typically observed α plate is closely correlated with the variations in nucleation rate and elastic strain energy induced by the external stress. However, it is still unclear how these changes result in the morphology transition of α precipitate. In this study, a multiphase-field and crystal plasticity integrated model was proposed to describe the coevolution of dynamic α precipitation and plastic deformation. It was found that the plasticity relaxes the elastic strain energy, retards lengthening but promotes the thickening of α precipitate. The elastic interaction between the applied stress and eigenstrain increases the elastic strain energy, promotes lengthening but has a weak effect on the thickening of α precipitate. The competition of these two factors determines the shape of α precipitates. In the case of a small nucleation rate, the promotion effect of the elastic interaction on lengthening of α precipitate always exceeds the retardation effect of plasticity, resulting in the formation of plate-like α. Increasing the nucleation rate under the same magnitude of external stress can induce a larger plastic strain within the soft β matrix in the initial growth stage, which results in the competitive advantage of the retardation effect of plasticity. Therefore, the lengthening of α precipitate decreases to a similar extent as its thickening, and the precipitate morphology changes into an equiaxed shape.

The problem that the conventional double-exponential transient current model (DE model) can overdrive the circuit, which leads to the overestimation of the soft error rate of the logic cell, is solved. Our work uses a new and accurate model for predicting the soft error rate that brings the soft error rate closer to the actual. The piecewise double-exponential transient current model (PDE model) is chosen, and the accuracy of the model is reflected using the Layout Awareness Single Event Multi Transients Soft Error Rate Calculation tool (LA-SEMT-SER tool). The model can characterize transient current pulses piecewise and limit the peak current magnitude to not exceed the conduction current. TCAD models are constructed from 28 nm process library and cell layouts. The transfer characteristic curves of devices are calibrated, and functional timing verification is performed to ensure the accuracy of the TCAD model. The experimental results show that the PDE model is not only more consistent with TCAD simulation than the DE model in modeling the single event transient currents of the device, but also that the SER calculated by the LA-SEMT-SER tool based on the PDE model has a smaller error than the SER calculated by the LA-SEMT-SER tool based on the DE model.

Camellia oil is valuable as an edible oil and serves as a base material for a range of high-value products. Camellia plants of significant economic importance, such as Camellia sinensis and Camellia oleifera, have been classified into sect. Thea and sect. Oleifera, respectively. Fatty acid desaturases play a crucial role in catalyzing the formation of double bonds at specific positions of fatty acid chains, leading to the production of unsaturated fatty acids and contributing to lipid synthesis. Comparative genomics results have revealed that expanded gene families in oil tea are enriched in functions related to lipid, fatty acid, and seed processes. To explore the function of the FAD gene family, a total of 82 FAD genes were identified in tea and oil tea. Transcriptome data showed the differential expression of the FAD gene family in mature seeds of tea tree and oil tea tree. Furthermore, the structural analysis and clustering of FAD proteins provided insights for the further exploration of the function of the FAD gene family and its role in lipid synthesis. Overall, these findings shed light on the role of the FAD gene family in Camellia plants and their involvement in lipid metabolism, as well as provide a reference for understanding their function in oil synthesis.

The basal plane dislocation (BPD) density is one of the most important defects affecting the application of SiC wafers. In this study, numerical simulations and corresponding experiments were conducted to investigate the influence of cooling processes, seed-bonding methods, and graphite crucible materials on the BPD density in an 8-inch N-type 4H-SiC single crystal grown by the physical vapor transport (PVT) method. The results showed that the BPD density could be effectively reduced by increasing the cooling rate, optimizing the seed-bonding method, and adopting a graphite crucible with a similar coefficient of thermal expansion as the SiC single crystal. The BPD density in the experiments showed that a high cooling rate reduced the BPD density from 4689 cm−2 to 2925 cm−2; optimization of the seed-bonding method decreased the BPD density to 1560 cm−2. The BPD density was further reduced to 704 cm−2 through the adoption of a graphite crucible with a smaller thermal expansion coefficient.

With the rapid development of semiconductor technology, the reduction in device operating voltage and threshold voltage has made integrated circuits more susceptible to the effects of particle radiation. Moreover, as process sizes decrease, the impact of charge sharing effects becomes increasingly severe, with soft errors caused by single event effects becoming one of the main causes of circuit failures. Therefore, the study of sensitivity evaluation methods for integrated circuits is of great significance for promoting the optimization of integrated circuit design, improving single event effect experimental methods, and enhancing the irradiation reliability of integrated circuits. In this paper, we first established a device model for the charge sharing effect and simulated it under reasonable conditions. Based on the simulation results, we then built a neural network model to predict the charge amounts in primary and secondary devices. We also propose a comprehensive automated method for calculating soft errors in unit circuits and validated it through TCAD simulations, achieving an error margin of 2.8–4.3%. This demonstrated the accuracy and effectiveness of the method we propose.

Bacillus cereus is an important opportunistic pathogen widely distributed in the environment. In this study, we reported the isolation and characterization of a B. cereus isolate, MB1, from the Challenger Deep of the Mariana Trench. MB1 is aerobic, motile, and able to form endospores. It possesses 5966 genes distributed on a circular chromosome and two plasmids. The MB1 genome contains 14 sets of 23S, 5S, and 16S ribosomal RNA operons, 106 tRNA genes, 4 sRNA genes, 12 genomic islands, 13 prophages, and 302 putative virulence genes, including enterotoxins and cytolysins. Infection studies showed that MB1 was able to cause acute and lethal infection in fish and mice, and was highly toxic to mammalian cells. MB1 induced, in a dose-dependent manner, pyroptotic cell death, characterized by activation of caspase-1, cleavage of gasdermin D, and release of IL-1β and IL-18. MB1 spores exhibited swimming and haemolytic capacity, but were severely attenuated in pathogenicity, which, however, was regained to the full extent when the spores germinated under suitable conditions. Taken together, these results provide new insights into the biological and pathogenic mechanism of deep sea B. cereus.

Abstract Objective To evaluate the clinical efficacy of the treatment of scoliosis with a pedicle screw system through paraspinal intermuscular approach (PIA). Methods This is a retrospective case series study. A total of 10 patients diagnosed with scoliosis had surgical indications and treated with a pedicle screw system in one‐stage posterior surgery by PIA from March 2013 to April 2015 at the First Hospital of Jilin University were enrolled in this study. The average age of the patients was 14.9 years, including one male and nine females. The operative information and surgical results, including Cobb angle correction, correction loss, global balance (including Frontal Plane Balance [FPB] and Sagittal Plane Balance [SPB]), and fusion rate were reviewed. Functional outcomes including visual analog scale (VAS) back pain score, leg pain score, and Scoliosis Research Society‐22 questionnaire (SRS‐22) were used to evaluate the quality of life of patients preoperatively and at last follow‐up. Results Each patient was followed up at least six times. The average follow‐up time was 43.2 months. Mean scoliosis and kyphosis improved from 68.5° ± 18.1°to 18.7° ± 11.8° and from 34.4° ± 17.9°to 24.0° ± 6.7°, respectively (p 0.05). During the follow‐up, mean coronal and sagittal correction loss was 1.4° ± 1.2°and 0.7° ± 0.8°, respectively (p > 0.05). Mean FPB improved from 32.7 to 11.7 mm (p 0.05). No dural tears were observed during the corrective surgery or wound infection or implant‐related complications. No pseudoarthrosis was identified according to the last follow‐up three‐dimensional (3D) CT scan. All the domains in SRS‐22 questionnaire show statistically significant improvement at the last follow‐up (p

To exploit the spectral-spatial information of hyperspectral image (HSI) and achieve higher the detection accuracy, a novel multiple spectral-spatial representation based on tensor decomposition method is proposed for HSI anomaly detection (AD) in this article. First, tensor decomposition is employed to separate the original HSI dataset into anomalous tensor from background tensor to, respectively, utilize the priors of background and anomaly. Then, the segmented smoothing prior is characterized by l0-l1 hybrid total variation regularization for the spatial dimensions of the background tensor, which can not only complement the image details but also fully consider the global spectral-spatial structure. Moreover, the low-rank prior is represented with truncated nuclear norm regularization for the spectral dimension of the background tensor to make full utilization of global information in the background and reduce data redundancy. Next, l2,1-norm regularization is applied to characterize the sparse prior for the anomaly tensor, which is beneficial to inscribe the local spectral structure of each image element. Finally, all these priors are fuses to model a unified convex optimization problem and obtain the final results by alternating direction method of multipliers, which can fully explore the discriminative spectral-spatial priors and improve the AD accuracy significantly. Experiments carried out on several real data indicate that our presented method reaches excellent performance in comparison to a few advanced AD methods.

Potassium is important for plant growth and crop yield. However, the effects of potassium (K+) deficiency on silage maize biomass yield and how maize shoot feedback mechanisms of K+ deficiency regulate whole plant growth remains largely unknown. Here, the study aims to explore the maize growth, transcriptional and metabolic responses of shoots to long-term potassium deficiency. Under the K+ insufficiency condition, the biomass yield of silage maize decreased. The transcriptome data showed that there were 922 and 1,107 differential expression genes in DH605 and Z58, respectively. In the two varieties, 390 differently expressed overlapping genes were similarly regulated. These genes were considered the fundamental responses to K+ deficiency in maize shoots. Many stress-induced genes are involved in transport, primary and secondary metabolism, regulation, and other processes, which are involved in K+ acquisition and homeostasis. Metabolic profiles indicated that most amino acids, phenolic acids, organic acids, and alkaloids were accumulated in shoots under K+ deficiency conditions and part of the sugars and sugar alcohols also increased. It revealed that putrescine and putrescine derivatives were specifically accumulated under the K+ deficiency condition, which may play a role in the feedback regulation of shoot growth. These results confirmed the importance of K+ on silage maize production and provided a deeper insight into the responses to K+ deficiency in maize shoots.

Based on Nickel isotope analysis of meteorites and terrestrial rocks, the authors suggest that the Bulk Silicate Earth has a sub-chondritic Nickel isotope composition. This signature is thought to result from the impact and accretion of a Mercury-like impactor which originated from the innermost Solar System.

Abstract Background Modulation of the host immune response by nematode parasites has been widely reported. Rhodaneses (thiosulfate: cyanide sulfurtransferases) are present in a wide range of organisms, such as archaea, bacteria, fungi, plants and animals. Previously, it was reported that a rhodanese homologue could be bound by goat peripheral blood mononuclear cells (PBMCs) in vivo. Methods In the present study, we cloned and produced a recombinant rhodanese protein originating from Haemonchus contortus (rHCRD), a parasitic nematode of small ruminants. rHCRD was co-incubated with goat PBMCs to assess its immunomodulatory effects on proliferation, apoptosis and cytokine secretion. Results We verified that the natural HCRD protein localized predominantly to the bowel wall and body surface of the parasite. We further demonstrated that serum produced by goats artificially infected with H. contortus successfully recognized rHCRD, which bound to goat PBMCs. rHCRD suppressed proliferation of goat PBMCs stimulated by concanavalin A but did not induce apoptosis in goat PBMCs. The production of TNF-α and IFN-γ decreased significantly, whereas secretion of IL-10 and TGF-β1 increased, in goat PBMCs after exposure to rHCRD. rHCRD also inhibited phagocytosis by goat monocytes. Moreover, rHCRD downregulated the expression of major histocompatibility complex (MHC)-II on goat monocytes in a dose-dependent manner, but did not alter MHC-I expression. Conclusions These results propose a possible immunomodulatory target that may help illuminate the interactions between parasites and their hosts at the molecular level and reveal innovative protein species as candidate drug and vaccine targets.

Abstract Background Methyltransferases (MTFs) are broad range of enzymes, which are ubiquitously expressed in diverse organisms ranging from bacteria to animals. MTFs proteins have been associated with various biological/cellular processes including transcriptional regulation, subcellular protein and RNA localization, signal transduction and DNA-damage repair. However, the role of MTFs in immune mechanism during host–parasite interaction has not been addressed yet. Results An open reading frame (764 bp) of methyltransferase-type 12 gene of H. contortus denoted as HcMTF-12, was successfully cloned using reverse transcriptase-polymerase chain reaction (RT-PCR) followed by prokaryotic expression in Escherichia coli BL21 (DE3 strain). The recombinant HcMTF-12 protein (rHcMTF-12) was about 47 kDa along with a fusion vector protein of 18 kDa. Immunoblot results identified the native protein MTF-12 with antibodies produced in rats against rHcMT-12, whereas rHcMTF-12 protein was recognized with sera of goat experimentally infected with H. contortus. Immunohistochemical analysis revealed that the native MTF-12 protein was mainly located in the periphery (cuticle) of parasite sections as well as within the pharynx and intestinal region. An immunofluorescence assay validated that rHcMTF-12 attached to the surface of goat PBMCs. Furthermore, the cytokines transcription of IL-2, IFN-γ and IL-4 transcripts of PBMCs incubated with rHcMTF-12 were enhanced in a dose-dependent manner. The secretion of TGF-β1 and IL-10 was significantly decreased. However, IL-6 production was not significantly different as compared to the control groups. Moreover, the migration activity and nitric oxide (NO) production by PBMCs were induced considerably, whereas the proliferation of PBMCs cells was negatively affected when incubated with the rHcMTF-12 protein. Conclusions Our findings suggest that HcMTF-12 significantly mediated the functions of PBMCs, and it might be a potential candidate for therapeutic interventions against haemonchosis.

Environmental quality is strongly coupling with the result of emissions reduction in most countries, while emissions reduction is influenced by the diversity of control policies, engineering facilities and development patterns. Further, local in which technical or policy that concern the control of target emissions will impact other pollutants emission. In China, the emissions of both CO2 and other pollutants increased substantially during the past decades. With the development of emissions reduction, it promoted a considerable risk of emissions spillovers caused by the present mechanism. In this study, we assessed the contributions of emissions spillovers to emission reductions by applying a more comprehensive indicator of pollutants, pollutant discharge fee (PDF), at the national, regional and provincial levels. The results shown positive and significant mutual effect between the PDF and CO2 emissions. That is, CO2 emission is significantly related with the process of other pollutants emissions reduction. Additionally, the positive correlation indicated further increases with the Five-Year Plan (FYP) cycle. Emissions spillovers have become an urgent risk for China. With the performance change, emissions reduction and environmental governance should heighten integration. Regional-level and provincial authorities must reconsider the matched emissions sustainability responsibilities and policies to avoid emissions spillovers and achieve meaningful emission controls. The study therefore suggested that a hierarchy of emission reductions, in the hope to promote emission reductions, improve environmental quality and contribute greenhouse gas reductions.

With the rapid development of industrialization and urbanization, emissions reduction is becoming an urgent mission for China. This work aims to review and analysis the effectiveness of emissions reduction in China based on the pollution discharge fee (PDF) and Five-year plan (FYP). The results show that, the emissions performance appeared significantly temporal performance changes under the current framework. Due to FYP and environment tax (ET) in parallel is an inevitable choice for environment and emission management, the connection and match among the existing emissions reduction policies are suggested to be enhanced. Four significant recommendations are raised: establishing dynamic economic regulation on emissions feedbacks, improving synchronization matching on information and data, increasing the audit on the technical applications and integrating more supporting policy with ET.

Brucella, a genus of bacteria that causes brucellosis, infects and threatens domestic animals, and humans in endemic areas. Presently, some live attenuated vaccines of Brucella are used to immunize livestock; however, these vaccines are pathogenic to humans, can provoke abortion when administered to pregnant livestock, and induce antibodies in vaccinated livestock that affect the diagnosis of field infection. It is, therefore, very important for improving the safety and immune protection effects of Brucella vaccine. Currently, recombinant protein-based subunit vaccines are considered promising safe and effective alternatives against brucellosis. Here, we separately expressed the recombinant Omp10-Omp28-L7/L12 proteins of Brucella using eukaryotic and prokaryotic expression systems, which were then used as immunogens for evaluating their immune responses. Taishan Pinus massoniana pollen polysaccharides (TPPPS), an already verified natural adjuvant, was utilized to evaluate the immune conditioning effect on the recombinant proteins. Antibody levels, spleen lymphocyte proliferation, percentages of CD4+ and CD8+ T cells, and cytokine secretion in mice were examined after three successive immunizations. The protective effects against Brucella challenge were also evaluated in mice, and used a live vaccine as a positive control. The results indicated that the immune responses of the recombinant Omp10-Omp28-L7/L12 protein groups were significantly higher than those of the PBS control group. The recombinant Omp10-Omp28-L7/L12 protein expressed in Pichia pastoris (P. pastoris) exhibited a slightly higher expression level and immunogenicity than that expressed in Escherichia coli (E. coli), and the Omp10-Omp28-L7/L12 (P. pastoris) + TPPPS group provided the most pronounced immune effect. The protective results showed that the recombinant Omp10-Omp28-L7/L12 proteins expressed in the two expression systems had significantly better protective effects against Brucella melitensis challenge compared with the negative control, and the addition of TPPPS adjuvant could significantly improve the protective effects of subunit vaccines. However, we also noticed that all of the evaluated subunit vaccines induced less protection than the B. melitensis M5 live vaccine. These results indicate that the combination of recombinant Omp10-Omp28-L7/L12 antigen and TPPPS adjuvant shows potential as an effective brucellosis subunit vaccine, and P. pastoris is a preferred expression system to prepare this recombinant subunit antigen.

Abstract Background The molecular structure of Fasciola gigantica 14-3-3 protein has been characterized. However, the involvement of this protein in parasite pathogenesis remains elusive and its effect on the functions of innate immune cells is unknown. We report on the cloning and expression of a recombinant F. gigantica 14-3-3 epsilon protein (rFg14-3-3e), and testing its effects on specific functions of goat peripheral blood mononuclear cells (PBMCs). Methods rFg14-3-3e protein was expressed in Pichia pastoris. Western blot and immunofluorescence assay (IFA) were used to examine the reactivity of rFg14-3-3e protein to anti-F. gigantica and anti-rFg14-3-3e antibodies, respectively. Various assays were used to investigate the stimulatory effects of the purified rFg14-3-3e protein on specific functions of goat PBMCs, including cytokine secretion, proliferation, migration, nitric oxide (NO) production, phagocytosis, and apoptotic capabilities. Potential protein interactors of rFg14-3-3e were identified by querying the databases Intact, String, BioPlex and BioGrid. A Total Energy analysis of each of the identified interaction was performed. Gene Ontology (GO) enrichment analysis was conducted using Funcassociate 3.0. Results Sequence analysis revealed that rFg14-3-3e protein had 100% identity to 14-3-3 protein from Fasciola hepatica. Western blot analysis showed that rFg14-3-3e protein is recognized by sera from goats experimentally infected with F. gigantica and immunofluorescence staining using rat anti-rFg14-3-3e antibodies demonstrated the specific binding of rFg14-3-3e protein to the surface of goat PBMCs. rFg14-3-3e protein stimulated goat PBMCs to produce interleukin-10 (IL-10) and transforming growth factor beta (TGF-β), corresponding with low levels of IL-4 and interferon gamma (IFN-γ). Also, this recombinant protein promoted the release of NO and cell apoptosis, and inhibited the proliferation and migration of goat PBMCs and suppressed monocyte phagocytosis. Homology modelling revealed 65% identity between rFg14-3-3e and human 14-3-3 protein YWHAE. GO enrichment analysis of the interacting proteins identified terms related to apoptosis, protein binding, locomotion, hippo signalling and leukocyte and lymphocyte differentiation, supporting the experimental findings. Conclusions Our data suggest that rFg14-3-3e protein can influence various cellular and immunological functions of goat PBMCs in vitro and may be involved in mediating F. gigantica pathogenesis. Because of its involvement in F. gigantica recognition by innate immune cells, rFg14-3-3e protein may have applications for development of diagnostics and therapeutic interventions.

Although most of deep-learning-based hyperspectral image (HSI) classification methods achieve great performance, there still remains a challenge to utilize small-size training samples to remarkably enhance the classification accuracy. To tackle this challenge, a novel two-branch spectral–spatial-feature attention network (TSSFAN) for HSI classification is proposed in this paper. Firstly, two inputs with different spectral dimensions and spatial sizes are constructed, which can not only reduce the redundancy of the original dataset but also accurately explore the spectral and spatial features. Then, we design two parallel 3DCNN branches with attention modules, in which one focuses on extracting spectral features and adaptively learning the more discriminative spectral channels, and the other focuses on exploring spatial features and adaptively learning the more discriminative spatial structures. Next, the feature attention module is constructed to automatically adjust the weights of different features based on their contributions for classification to remarkably improve the classification performance. Finally, we design the hybrid architecture of 3D–2DCNN to acquire the final classification result, which can significantly decrease the sophistication of the network. Experimental results on three HSI datasets indicate that our presented TSSFAN method outperforms several of the most advanced classification methods.

Abstract Background Haemonchosis is a disease of the small ruminant caused by a nematode parasite Haemonchus contortus, and it is most important and alarming challenges to the small ruminant’s production. The infection of the H. contortus could cause high economic losses worldwide. H. contortus is a blood feeding parasite which penetrates into the abomasal mucosa to feed the blood of the host and causing the anemia and decreased total plasma protein. Modulation and suppression of the immune response of the host by nematode parasites have been reported extensively, and the cysteine protease inhibitor (cystatin) is identified as one of the major immunomodulators. Methods The recombinant protein of HCcyst-3 was expressed in a histidine-tagged fusion soluble form in Escherichia coli, and its inhibitory activity against cathepsin L, B, as well as papain, were identified by fluorogenic substrate analysis. Native HCcyst-3 protein was localized by an Immunohistochemical test. The immunomodulatory effects of HCcyst-3 on cytokine secretion, MHC molecule expression, NO production and phagocytosis were observed by co-incubation of rHCcyst-3 with goat monocytes. Results We cloned and produced recombinant cystatin protein from H. contortus (rHCcyst-3) and investigated its immunomodulatory effects on goat monocyte. The rHCcyst-3 protein is biologically functional as shown by its ability to inhibit the protease activity of cathepsin L, cathepsin B, and papain. The immunohistochemical test demonstrated that the native HCcyst-3 protein was predominantly localized at the body surface and internal surface of the worm’s gut. We demonstrated that rHCcyst-3 could be distinguished by antisera from goat experimentally infected with H. contortus and could uptake by goat monocytes. The results showed that the engagement of rHCcyst-3 decreased the production of TNF-α, IL-1β and IL-12p40. However, it significantly increased the secretion of IL-10 and TGF-β1 in goat monocytes. After rHCcyst-3 exposure, the expression of MHC-II on goat monocytes was restricted. Moreover, rHCcyst-3 could upregulate LPS induced NO production of goat monocytes. Phagocytotic assay by FITC-dextran internalization showed that rHCcyst-3 inhibited the phagocytosis of goat monocytes. Conclusions Our results suggested that the recombinant cystatin from H. contortus (rHCcyst-3) significantly modulated goat monocyte function in multiple aspects.

Abstract Background Arginine kinase (AK), an important member of phosphagen kinase family has been extensively studied in various vertebrates and invertebrates. Immunologically, AKs are important constituents of different body parts, involved in various biological and cellular functions, and considered as immune-modulator and effector for pro-inflammatory cytokines. However, immunoregulatory changes of host cells triggered by AK protein of Haemonchus contortus, a parasitic nematode of ruminants, are still unknown. The current study was focused on cloning and characterisation of Hc-AK, and its regulatory effects on cytokines level, cell migration, cell proliferation, nitric oxide production and apoptosis of goat peripheral blood mononuclear cells (PBMCs) were observed. Methods The full-length sequence of the Hc-AK gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and sub-cloned into the prokaryotic expression vector pET-32a. The biochemical characteristics of recombinant protein Hc-AK, which was purified by affinity chromatography, were performed based on the enzymatic assay. Binding of rHc-AK with PBMCs was confirmed by immunofluorescence assay (IFA). Immunohistochemical analysis was used to detect localisation of Hc-AK within adult worms sections. The immunoregulatory effects of rHc-AK on cytokine secretions, cell proliferation, cell migration, nitric oxide production and apoptosis were determined by co-incubation of rHc-AK with goat PBMCs. Results The full-length ORF (1080 bp) of the Hc-AK gene was successfully cloned, and His-tagged AK protein was expressed in the Escherichia coli strain BL21. The recombinant protein of Hc-AK (rHc-AK) was about 58.5 kDa together with the fused vector protein of 18 kDa. The biochemical assay showed that the protein encoded by the Hc-ak exhibited enzymatic activity. Western blot analysis confirmed that the rHc-AK was recognised by the sera from rat (rat-antiHc-AK). The IFA results showed that rHc-AK could bind on the surface of goat PBMCs. Immunohistochemically, Hc-AK was localised at the inner and outer membrane as well as in the gut region of adult worms. The binding of rHc-AK to host cells increased the levels of IL-4, IL-10, IL-17, IFN-γ, nitric oxide (NO) production and cell apoptosis of goat PBMCs, whereas, TGF-β1 levels, cell proliferation and PBMCs migration were significantly decreased in a dose dependent manner. Conclusions Our findings suggested that rHc-AK is an important excretory and secretory (ES) protein involved in host immune responses and exhibit distinct immunomodulatory properties during interaction with goat PBMCs.

Hollow sphere (Ba(0.6)Sr(0.4))TiO(3) (HS-BST) powders with sub-micron size and different shell thickness were synthesised through template-assisted method. HS-BST/poly(vinylidene fluoride) (PVDF) composites were prepared using tape casting method. The effects of particle size and shell thickness of HS-BST on the microstructure, dielectric tunability, and energy storage properties of HS-BST/PVDF composites were investigated. Results showed that all HS-BST particles were uniformly dispersed in PVDF matrix without agglomeration. With decreasing the size of HS-BST particles, the dielectric constant and loss of HS-BST/PVDF composites decreased, while the breakdown strength initially decreased and then increased. In addition, the theoretical model of the dielectric tunability of inorganic/organic system related to the shape factor (n) of inorganic particles was used to simulate and n value of 0-dimensional spherical particles was estimated. For HS-BST/PVDF composite, the n in the model was 2–2.5, corresponding to HS-BST particles with a diameter ranging from 0.44 to 2.57 μm, and a shell thickness ranging from 40 to 500 nm.

Crustin is a type of antimicrobial peptide and plays an important role in the innate immunity of arthropods. We report here the identification and characterization of a crustin (named Crus1) from the shrimp Rimicaris sp. inhabiting the deep-sea hydrothermal vent in Manus Basin (Papua New Guinea). Crus1 shares the highest identity (51.76%) with a Type I crustin of Penaeus vannamei and possesses a whey acidic protein (WAP) domain, which contains eight cysteine residues that form the conserved ‘four-disulfide core’ structure. Recombinant Crus1 (rCrus1) bound to peptidoglycan and lipoteichoic acid, and effectively killed Gram-positive bacteria in a manner that was dependent on pH, temperature, and disulfide linkage. rCrus1 induced membrane leakage and structure damage in the target bacteria, but had no effect on bacterial protoplasts. Serine substitution of each of the 8 Cys residues in the WAP domain did not affect the bacterial binding capacity but completely abolished the bactericidal activity of rCrus1. These results provide new insights into the characteristic and mechanism of the antimicrobial activity of deep sea crustins.

The importance of the gut microbiota (GM) of animals is widely acknowledged because of its pivotal roles in metabolism, immunity, and health maintenance. The level of health can be reflected by the dynamic distribution of GM. In this study, high-throughput sequencing of the bacterial 16S rRNA gene was used to compare the microbial populations from feces in healthy and diarrheic kids, which reflected the dynamic shift of microbiota in kids and investigated differences from adult healthy goats. Healthy kids and goats not only displayed higher species richness but also exhibited higher bacterial diversity than diarrheic kids based on the results of the operational taxonomic unit analysis, alpha diversity, and beta diversity. Firmicutes and Bacteroidetes were the most dominant phyla in all samples. At the genus level, the differences in diversity and abundance between diarrheic kids and the other two groups were gradually observed. In the diarrheic kid intestine, Bacteroides remained the dominant species, and the proportion of Clostridium_sensu_stricto_1 and Paeniclostridium increased, whereas Rikenellaceae_RC9_gut_group, Ruminococcaceae_UCG-005, and Christensenellaceae_R-7_group were significantly reduced. The results showed the differences of GM in diarrheic kids and healthy kids were significant while in kids and goats were not obvious. Differences in the composition of intestinal microbiota may not be the cause of diarrhea, and some changes of bacterial richness may guide our interpretation of diarrhea. This study is the first to investigate the distribution of GM in Boer goats with different ages and health states. Furthermore, this study will provide a theoretical basis for the establishment of a prevention and treatment system for goat diarrhea.

Serine/threonine-protein phosphatases (STPs), as integral constituents of parasitic excretory/secretory proteins, are assumed to be released during the host–parasite interactions. However, knowledge about these phosphatases and their immunoregulatory and immune protective efficiencies with host peripheral blood mononuclear cells (PBMCs) is scant. In this study, an open reading frame of STP from Haemonchus contortus designated as HcSTP-1 was amplified and cloned using reverse-transcription-polymerase chain reaction (RT-PCR) method. The 951-bp nucleotides sequence was encoded to a protein of 316 amino acid residues, conserved in characteristics motifs GDXHG, GDYVDRG, GNHE, HGG, RG, and H. The HcSTP-1 protein was detected at approximately 35 kDa as recombinant protein fused in an expression vector system and resolved on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Immunohistochemically, HcSTP-1 was found to be localized in both male and female adult worm sections. Using immunofluorescence assay, the binding activity of rHcSTP-1 was confirmed on surface of goat PBMCs, which resulted in expression of multiple cytokines and various immunoregulatory activities in vitro. The RT-PCR results showed that mRNA level of interleukin-2, TGF-β1, IFN-γ, and IL-17 (with 10 µg/ml) was upregulated and IL-10 was decreased. However, IL-6 showed no change after PBMCs incubated with rHcSTP-1 protein. Further functional analysis showed that migratory activity of cells, intracellular nitrite production (NO), and apoptotic efficiency of PBMCs were elevated at significant level, whereas the proliferation of goat PBMCs and monocytes-associated major histocompatibility complex (MHC)-I and MHC-II expressions were decreased significantly at concentration-dependent fashion. Our results showed that the HcSTP-1 protein engaged in vital suppressive regulatory roles on host immune cells, which might represent a potential molecular target for controlling H. contortus infection in future.

Abstract Background Ticks and tick-borne diseases affect animal and human health worldwide and cause significant economic losses in the animal industry. Functional molecular research is important to understand the biological characteristics of ticks at the molecular level. Enzymes and enzyme inhibitory molecules play very important roles in tick physiology, and the cystatins are tight-binding inhibitors of papain-like cysteine proteases. To this end, a novel cystatin, designated RHcyst-1, was isolated from the tick Rhipicephalus haemaphysaloides. Methods The full-length gene of RHcyst-1 was cloning by RACE. The recombinant protein of RHcyst-1 was expressed in a glutathione S-transferase (GST)-fused soluble form in Escherichia coli, and its inhibitory activity against cathepsin L, B, C, H, and S, as well as papain, was identified by fluorogenic substrate analysis. Expression analysis of RHcyst-1 at different tick stages was performed by quantitative reverse transcription - PCR (qRT-PCR). An RNAi experiment for RHcyst-1 was performed to determine its function for tick physiology. Results The full-length cDNA of RHcyst-1 is 471 bp, including an intact open reading frame encoding an expected protein of 98 amino acids, without a signal peptide, having a predicted molecular weight of ~11 kDa and an isoelectric point of 5.66. A sequence analysis showed that it has significant homology with the known type 1 cystatins. The results of proteinase inhibition assays showed that rRHcyst-1 can effectively inhibit the six cysteine proteases’ enzyme activities. An investigation of the RHcyst-1 genes’ expression profile showed that it was more richly transcribed in the embryo (egg) stage. A disruption of the RHcyst-1 gene showed a significant decrease in the rate of tick hatching. Conclusions Our results suggested that RHcyst-1 may be involved in the early embryonic development of ticks.

Radio frequency (RF) technique, for its better penetrability over traditional techniques such as infrared or ultrasound, is widely used for indoor localization and tracking. In this paper, three novel measurements, point decision accuracy, path matching error and wrong jumping ratio, are firstly defined to express the localization efficiency. Then, a novel RSSI-based smooth localization (RSL) algorithm is designed, implemented, and evaluated on the WiFi networks. The tree-based mechanism determines the current position and track of the entity by assigning the weights and accumulative weights for all collected RSSI information of reference points so as to make the localization smooth. The evaluation results indicate that the proposed algorithm brings better localization smoothness of reducing 10% path matching error and 30% wrong jumping ratio over the RADAR system.

The effectiveness of facial expression recognition (FER) algorithms hinges on the model’s quality and the availability of a substantial amount of labeled expression data. However, labeling large datasets demands significant human, time, and financial resources. Although active learning methods have mitigated the dependency on extensive labeled data, a cold-start problem persists in small to medium-sized expression recognition datasets. This issue arises because the initial labeled data often fails to represent the full spectrum of facial expression characteristics. This paper introduces an active learning approach that integrates uncertainty estimation, aiming to improve the precision of facial expression recognition regardless of dataset scale variations. The method is divided into two primary phases. First, the model undergoes self-supervised pre-training using contrastive learning and uncertainty estimation to bolster its feature extraction capabilities. Second, the model is fine-tuned using the prior knowledge obtained from the pre-training phase to significantly improve recognition accuracy. In the pretraining phase, the model employs contrastive learning to extract fundamental feature representations from the complete unlabeled dataset. These features are then weighted through a self-attention mechanism with rank regularization. Subsequently, data from the low-weighted set is relabeled to further refine the model’s feature extraction ability. The pre-trained model is then utilized in active learning to select and label information-rich samples more efficiently. Experimental results demonstrate that the proposed method significantly outperforms existing approaches, achieving an improvement in recognition accuracy of 5.09% and 3.82% over the best existing active learning methods, Margin, and Least Confidence methods, respectively, and a 1.61% improvement compared to the conventional segmented active learning method. [ABSTRACT FROM AUTHOR]