The rifampicin-resistant strain E9-302 of
strain 669 (WT) was generated by continuous passage on BHI agar plates containing increasing concentrations of rifampicin. E9-302 was attenuated ...significantly by 119 times to zebrafish
compared to WT in terms of the 50% lethal dose (LD
). Zebrafish vaccinated with E9-302 via intraperitoneal (IP) injection at a dose of 1 × 10
CFU/fish had relative percentage survival (RPS) rates of 85.7% when challenged with wild-type
via IP 14 days post-vaccination (dpv). After 14 days of primary vaccination with E9-302 via immersion (IM) at a dose of 4 × 10
CFU/ml, a booster IM vaccination with E9-302 at a dose of 2 × 10
CFU/ml exhibited 65.2% RPS against challenge with wild-type
via IP 7 days later. These results indicated that the rifampicin-resistant attenuated strain E9-302 had potential as a live vaccine against
infection. A previously unreported amino acid site change at position 142 of the RNA polymerase (RNAP) β subunit encoded by the gene
associated with rifampicin resistance was identified. Analysis of the whole-genome sequencing results revealed multiple missense mutations in the virulence-related genes
and
in E9-302 compared with WT, and a 189 bp mismatch in one gene, whose coding product was highly homologous to glycosyltransferase family 39 protein. This study preliminarily explored the molecular mechanism underlying the virulence attenuation of rifampicin-resistant strain E9-302 and provided a new target for the subsequent study of the pathogenic mechanism of
.
This paper presents a comprehensive study based on multiphase-seepage and wellbore multiphase-flow theories. It establishes a model for calculating the rate of gas intrusion that considers various ...factors, including formation pore permeability, bottomhole pressure difference, rheology of the drilling fluid, and surface tension. Experiments were conducted to investigate the mechanism of gas intrusion under shut-in conditions, and the experimental results were employed to validate the reliability of the proposed method for calculating the gas intrusion rate. Furthermore, this research explores the transportation rates of single bubbles and bubble clusters in drilling fluid under shut-in conditions. Additionally, empirical expressions were derived for the drag coefficient for single bubbles and bubble clusters in the wellbore. These expressions can be used to calculate gas transportation rates for various equivalent radii of single bubbles and bubble clusters. The initial bubble size of intrusive gas, the transportation speed of intrusive gas in the wellbore, the rate of gas intrusion, and variations in the wellbore pressure after gas intrusion were analyzed. Additionally, a method was developed to calculate the rising velocity of bubble clusters in water based on experimental results. The study reveals that the average bubble size in the bubble cluster is significantly smaller than the size of single bubbles generated from the orifice. When the viscosity of the drilling fluid is low, the transportation velocity of the bubble cluster exhibits a positive correlation with the average bubble diameter. When the average bubble diameter exceeds 1 mm, the bubble velocity no longer varies with changes in the bubble-cluster diameter. The research results provide theoretical support for wellbore pressure prediction and pressure control under shutdown conditions.
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Understanding agents’ motion behaviors under complex scenes is crucial for intelligent autonomous moving systems (like delivery robots and self-driving cars). It is challenging duo to the inherent ...uncertain of future trajectories and the large variation in the scene layout. However, most recent approaches ignored or underutilized the scenario information. In this work, a Multi-Granularity Scenarios Understanding framework, MGSU, is proposed to explore the scene layout from different granularity. MGSU can be divided into three modules: (1) A coarse-grained fusion module uses the cross-attention to fuse the observed trajectory with the semantic information of the scene. (2) The inverse reinforcement learning module generates optimal path strategy through grid-based policy sampling and outputs multiple scene paths. (3) The fine-grained fusion module integrates the observed trajectory with the scene paths to generate multiple future trajectories. To fully explore the scene information and improve the efficiency, we present a novel scene-fusion Transformer, whose encoder is used to extract scene features and the decoder is used to fuse scene and trajectory features to generate future trajectories. Compared with the current state-of-the-art methods, our method decreases the ADE errors by 4.3% and 3.3% by gradually integrating different granularity of scene information on SDD and NuScenes, respectively. The visualized trajectories demonstrate that our method can accurately predict future trajectories after fusing scene information.
We previously developed and assessed the effectiveness of the attenuated
Streptococcus agalactiae
(Group B
Streptococcus
, GBS) strain WC1535 ∆Sia (with
neuA-D
gene cluster deletion) vaccine in ...tilapia (
Oreochromis niloticus
). In this study, we characterized the bacterial communities of the tilapia intestines by 16S rRNA high-throughput sequencing and assessed the serum antibody response, expression of immune-related genes, and histological changes following formalin-killed GBS vaccine (FKV) and the live attenuated vaccine ∆Sia (LAV). Results showed that FKV and LAV induced robust systemic and intestinal mucosal immune responses in tilapia without causing obvious pathological changes in the hindgut, spleen, and head kidney but exerted different effects on intestinal bacterial communities. The richness or diversity of the intestinal bacterial community of FKV tilapia showed no significant changes compared with that of the control fish (
p
> 0.05) at either day 21 post-initial vaccination (21 dpiv) or day 35 (day 14 after the second immunization) (35 dpiv). The community composition of FKV tilapia and controls was significantly similar, although the relative abundance of some genera was significantly altered. Relative to control fish, the gut ecosystem of LAV tilapia was significantly disturbed with a substantial increase in community diversity at 21 dpiv (
p
< 0.05) and a significant decrease at 35 dpiv in fish with high serum antibody response (ΔSia35H) (
p
< 0.05). However, there was no significant difference between ΔSia35H and ΔSia35L (low serum antibody response) fish (
p
> 0.05). Moreover, the community composition of LAV tilapia at 21 dpiv or 35 dpiv was considerably different from that of the controls. Particularly, GBS ∆Sia was found to be abundant in the intestine at 21 and 35 dpiv. This result suggested that the parenteral administration of the LAV (∆Sia) may also have the effect of oral vaccination in addition to the immune effect of injection vaccination. In addition, a significant correlation was found between the expression of immune-related genes and certain bacterial species in the intestinal mucosal flora. Our findings will contribute to a better understanding of the effects of inactivated and attenuated vaccines on gut microbiota and their relationship with the immune response.
Ship weather routing has always been an important issue in the research field of navigation, and many scholars have been devoted to this research for a long time. To study the route strategies of ...different shipping companies, this paper proposes an improved multi-objective ant colony optimization (IMACO) algorithm based on the Technique for Order Preference by Similarity to Ideal Solution (TOPSIS). It can comprehensively consider ship navigation risk and fuel consumption cost under complex sea conditions. First, the grid method is used to model the marine environment. Then, we calculate the fuel consumption and the ship navigation risk of each grid and use the TOPSIS method to evaluate these two indicators for each grid. The results show that due to the different strictness of navigation management requirements of different companies, different routes are selected in the same sea area at the same time. Compared with the single-objective ant colony optimization (SACO) algorithm, the algorithm proposed in this paper can more comprehensively and effectively solve the problem of route strategy selection of shipping companies, which has great practical significance for ship operations management.
Tetraspanins, a superfamily of membrane proteins, mediate diverse biological processes through tetraspanin-enriched microdomains in the plasma membrane. However, how their cell-surface presentation ...is controlled remains unclear. To identify the regulators of tetraspanin trafficking, we conduct sequential genome-wide loss-of-function CRISPR-Cas9 screens based on cell-surface expression of a tetraspanin member, TSPAN8. Several genes potentially involved in endoplasmic reticulum (ER) targeting, different biological processes in the Golgi apparatus, and protein trafficking are identified and functionally validated. Importantly, we find that biantennary N-glycans generated by MGAT1/2, but not more complex glycan structures, are important for cell-surface tetraspanin expression. Moreover, we unravel that SPPL3, a Golgi intramembrane-cleaving protease reported previously to act as a sheddase of multiple glycan-modifying enzymes, controls cell-surface tetraspanin expression through a mechanism associated with lacto-series glycolipid biosynthesis. Our study provides critical insights into the molecular regulation of cell-surface presentation of tetraspanins with implications for strategies to manipulate their functions, including cancer cell invasion.
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•Identifying key regulators of tetraspanin trafficking by CRISPR-Cas9 screens•The role of N-glycosylation in cell-surface presentation of tetraspanins•Control of cell-surface tetraspanin expression by a SPPL3/B3GNT5 axis•MGAT1/2 and SPPL3 are potential targets for blocking cancer cell invasion
Using sequential genome-wide loss-of-function CRISPR-Cas9 screens, Yang et al. identify key molecular regulators of cell-surface tetraspanin presentation. They unravel that N-glycosylation modification and biosynthesis of lacto-series glycolipids control the internalization of tetraspanins. Their findings constitute an avenue for modulating the diverse functions of tetraspanins, including cancer cell invasion.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
Tetraspanins, a superfamily of small integral membrane proteins, are characterized by four transmembrane domains and conserved protein motifs that are configured into a unique molecular topology and ...structure in the plasma membrane. They act as key organizers of the plasma membrane, orchestrating the formation of specialized microdomains called "tetraspanin-enriched microdomains (TEMs)" or "tetraspanin nanodomains" that are essential for mediating diverse biological processes. TSPAN8 is one of the earliest identified tetraspanin members. It is known to interact with a wide range of molecular partners in different cellular contexts and regulate diverse molecular and cellular events at the plasma membrane, including cell adhesion, migration, invasion, signal transduction, and exosome biogenesis. The functions of cell-surface TSPAN8 are governed by ER targeting, modifications at the Golgi apparatus and dynamic trafficking. Intriguingly, limited evidence shows that TSPAN8 can translocate to the nucleus to act as a transcriptional regulator. The transcription of
is tightly regulated and restricted to defined cell lineages, where it can serve as a molecular marker of stem/progenitor cells in certain normal tissues as well as tumors. Importantly, the oncogenic roles of TSPAN8 in tumor development and cancer metastasis have gained prominence in recent decades. Here, we comprehensively review the current knowledge on the molecular characteristics and regulatory mechanisms defining TSPAN8 functions, and discuss the potential and significance of TSPAN8 as a biomarker and therapeutic target across various epithelial cancers.
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Symbiotic microorganisms in the digestive and circulatory systems are found in various crustaceans, and their essential roles in crustacean health, nutrition, and disease have attracted considerable ...interest. Although the intestinal microbiota of the Chinese mitten crab (
) has been extensively studied, information on the symbiotic microbiota at various sites of this aquatic economic species, particularly the hepatopancreas and hemolymph, is lacking. This study aimed to comprehensively characterize the hemolymph, hepatopancreas, and intestinal microbiota of Chinese mitten crab through the high-throughput sequencing of the 16S rRNA gene. Results showed no significant difference in microbial diversity between the hemolymph and hepatopancreas (Welch
-test;
> 0.05), but their microbial diversity was significantly higher than that in the intestine (
< 0.05). Distinct differences were found in the structure, composition, and predicted function of the symbiotic microbiota at these sites. At the phylum level, the hemolymph and hepatopancreas microbiota were dominated by Proteobacteria, Firmicutes, and Acidobacteriota, followed by Bacteroidota and Actinobacteriota, whereas the gut microbiota was mainly composed of Firmicutes, Proteobacteria, and Bacteroidota. At the genus level,
,
, and
were dominant in the hepatopancreas;
,
and
were dominant in the intestine;
, norank_Vicinamibacterales, and
were relatively high-abundance genera in the hemolymph. The composition and abundance of symbiotic microbiota in the hemolymph and hepatopancreas were extremely similar (
> 0.05), and no significant difference in functional prediction was found (
> 0.05). Comparing the hemolymph in the intestine and hepatopancreas, the hemolymph had lower variation in bacterial composition among individuals, having a more uniform abundance of major bacterial taxa, a smaller coefficient of variation, and the highest proportion of shared genera. Network complexity varied greatly among the three sites. The hepatopancreas microbiota was the most complex, followed by the hemolymph microbiota, and the intestinal microbiota had the simplest network. This study revealed the taxonomic and functional characteristics of the hemolymph, hepatopancreas, and gut microbiota in Chinese mitten crab. The results expanded our understanding of the symbiotic microbiota in crustaceans, providing potential indicators for assessing the health status of Chinese mitten crab.
After millions of years of coevolution, symbiotic microbiota has become an integral part of the host and plays an important role in host immunity, metabolism, and health. Vaccination, as an effective ...means of preventing infectious diseases, has been playing a vital role in the prevention and control of human and animal diseases for decades. However, so far, minimal is known about the effect of vaccination on fish symbiotic microbiota, especially mucosal microbiota, and its correlation with intestinal metabolism remains unclear.
Here we reported the effect of an inactivated bivalent Aeromonas hydrophila/Aeromonas veronii vaccine on the symbiotic microbiota and its correlation with the intestinal metabolism of farmed adult Nile tilapia (Oreochromis niloticus) by 16S rRNA gene high-throughput sequencing and gas chromatography-mass spectrometry metabolomics.
Results showed that vaccination significantly changed the structure, composition, and predictive function of intestinal mucosal microbiota but did not significantly affect the symbiotic microbiota of other sites including gill mucosae, stomach contents, and stomach mucosae. Moreover, vaccination significantly reduced the relative abundance values of potential opportunistic pathogens such as Aeromonas, Escherichia-Shigella, and Acinetobacter in intestinal mucosae. Combined with the enhancement of immune function after vaccination, inactivated bivalent Aeromonas vaccination had a protective effect against the intestinal pathogen infection of tilapia. In addition, the metabolite differential analysis showed that vaccination significantly increased the concentrations of carbohydrate-related metabolites such as lactic acid, succinic acid, and gluconic acid but significantly decreased the concentrations of multiple lipid-related metabolites in tilapia intestines. Vaccination affected the intestinal metabolism of tilapia, which was further verified by the predictive function of intestinal microbiota. Furthermore, the correlation analyses showed that most of the intestinal differential microorganisms were significantly correlated with intestinal differential metabolites after vaccination, confirming that the effect of vaccination on intestinal metabolism was closely related to the intestinal microbiota.
In conclusion, this paper revealed the microbial and metabolic responses induced by inactivated vaccination, suggesting that intestinal microbiota might mediate the effect of vaccination on the intestinal metabolism of tilapia. It expanded the novel understanding of vaccine protective mechanisms from microbial and metabolic perspectives, providing important implications for the potential influence of vaccination on human intestinal microbiota and metabolism. Video Abstract.
Polyethylenimine (PEI) has attracted much attention as a DNA condenser, but its toxicity and non-specific targeting limit its potential. To overcome these limitations, Antheraea pernyi silk fibroin ...(ASF), a natural protein rich in arginyl-glycyl-aspartic acid (RGD) peptides that contains negative surface charges in a neutral aqueous solution, was used to coat PEI/DNA complexes to form ASF/PEI/DNA ternary complexes. Coating these complexes with ASF caused fewer surface charges and greater size compared with the PEI/DNA complexes alone. In vitro transfection studies revealed that incorporation of ASF led to greater transfection efficiencies in both HEK (human embryonic kidney) 293 and HCT (human colorectal carcinoma) 116 cells, albeit with less electrostatic binding affinity for the cells. Moreover, the transfection efficiency in the HCT 116 cells was higher than that in the HEK 293 cells under the same conditions, which may be due to the target bonding affinity of the RGD peptides in ASF for integrins on the HCT 116 cell surface. This result indicated that the RGD binding affinity in ASF for integrins can enhance the specific targeting affinity to compensate for the reduction in electrostatic binding between ASF-coated PEI carriers and cells. Cell viability measurements showed higher cell viability after transfection of ASF/PEI/DNA ternary complexes than after transfection of PEI/DNA binary complexes alone. Lactate dehydrogenase (LDH) release studies further confirmed the improvement in the targeting effect of ASF/PEI/DNA ternary complexes to cells. These results suggest that ASF-coated PEI is a preferred transfection reagent and useful for improving both the transfection efficiency and cell viability of PEI-based nonviral vectors.
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