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1.
Sci Rep ; 14(1): 13192, 2024 06 08.
Article in English | MEDLINE | ID: mdl-38851826

ABSTRACT

Water eutrophication has emerged as a pressing concern for massive algal blooms, and these harmful blooms can potentially generate harmful toxins, which can detrimentally impact the aquatic environment and human health. Consequently, it is imperative to identify a safe and efficient approach to combat algal blooms to safeguard the ecological safety of water. This study aimed to investigate the procedure for extracting total flavonoids from Z. bungeanum residue and assess its antioxidant properties. The most favorable parameters for extracting total flavonoids from Z. bungeanum residue were a liquid-solid ratio (LSR) of 20 mL/g, a solvent concentration of 60%, an extraction period of 55 min, and an ultrasonic temperature of 80 °C. Meanwhile, the photosynthetic inhibitory mechanism of Z. bungeanum residue extracts against M. aeruginosa was assessed with a particular focus on the concentration-dependent toxicity effect. Z. bungeanum residue extracts damaged the oxygen-evolving complex structure, influenced energy capture and distribution, and inhibited the electron transport of PSII in M. aeruginosa. Furthermore, the enhanced capacity for ROS detoxification enables treated cells to sustain their photosynthetic activity. The findings of this study hold considerable relevance for the ecological management community and offer potential avenues for the practical utilization of resources in controlling algal blooms.


Subject(s)
Flavonoids , Microcystis , Photosynthesis , Zanthoxylum , Flavonoids/chemistry , Flavonoids/isolation & purification , Flavonoids/pharmacology , Zanthoxylum/chemistry , Plant Extracts/pharmacology , Plant Extracts/chemistry , Antioxidants/pharmacology , Antioxidants/chemistry , Antioxidants/isolation & purification , Allelopathy , Harmful Algal Bloom , Reactive Oxygen Species/metabolism , Photosystem II Protein Complex/metabolism
2.
Science ; 384(6701): eadk5382, 2024 Jun 14.
Article in English | MEDLINE | ID: mdl-38870290

ABSTRACT

Polycystic ovary syndrome (PCOS), a prevalent reproductive disorder in women of reproductive age, features androgen excess, ovulatory dysfunction, and polycystic ovaries. Despite its high prevalence, specific pharmacologic intervention for PCOS is challenging. In this study, we identified artemisinins as anti-PCOS agents. Our finding demonstrated the efficacy of artemisinin derivatives in alleviating PCOS symptoms in both rodent models and human patients, curbing hyperandrogenemia through suppression of ovarian androgen synthesis. Artemisinins promoted cytochrome P450 family 11 subfamily A member 1 (CYP11A1) protein degradation to block androgen overproduction. Mechanistically, artemisinins directly targeted lon peptidase 1 (LONP1), enhanced LONP1-CYP11A1 interaction, and facilitated LONP1-catalyzed CYP11A1 degradation. Overexpression of LONP1 replicated the androgen-lowering effect of artemisinins. Our data suggest that artemisinin application is a promising approach for treating PCOS and highlight the crucial role of the LONP1-CYP11A1 interaction in controlling hyperandrogenism and PCOS occurrence.


Subject(s)
ATP-Dependent Proteases , Artemisinins , Cholesterol Side-Chain Cleavage Enzyme , Mitochondrial Proteins , Polycystic Ovary Syndrome , Animals , Female , Humans , Mice , Rats , Androgens/metabolism , Artemisinins/therapeutic use , Artemisinins/pharmacology , Cholesterol Side-Chain Cleavage Enzyme/metabolism , Cholesterol Side-Chain Cleavage Enzyme/genetics , Disease Models, Animal , Hyperandrogenism/drug therapy , Hyperandrogenism/metabolism , Mitochondrial Proteins/metabolism , Mitochondrial Proteins/genetics , Ovary/drug effects , Ovary/metabolism , Polycystic Ovary Syndrome/drug therapy , Proteolysis , Mice, Inbred C57BL , Young Adult , Adult , Rats, Sprague-Dawley , ATP-Dependent Proteases/genetics , ATP-Dependent Proteases/metabolism
3.
Opt Express ; 30(13): 23295-23304, 2022 Jun 20.
Article in English | MEDLINE | ID: mdl-36225013

ABSTRACT

We report a whispering gallery mode (WGM)-based fiber optofluidic laser (FOFL), in which rhodamine B (RhB) in an aqueous surfactant solution of sodium dodecylbenzene sulfonate (SDBS) is used as the laser gain medium. Here, the role of SDBS is to scatter the RhB dye molecules to effectively prevent its self-association in the aqueous solution. Therefore, the fluorescence quantum yield of the used RhB dye is improved due to the enhanced solubilization, which results in a low lasing threshold of ∼2.2 µJ/mm2 when the concentration of SDBS aqueous solution reaches up to 20 mM, on par with or even better than most of the optofluidic dye lasers using RhB as the gain medium in an organic solution. We then establish a model of solubilization capacity of SDBS micelles, which successfully addresses the mechanisms of dye-surfactant interactions in the proposed FOFL system. We further apply this FOFL platform to the case of concentration sensing of the used SDBS, which exhibits a 2-order-of-magnitude improvement in sensitivity compared to the fluorescence measurement due to the signal amplification inherent to the lasing process. The proposed FOFL platform in combination with surfactant solubilization gain medium in an aqueous solution promises to enable chip-scale coherent light sources for various environmental and bio-chemical sensing applications.

4.
Angew Chem Int Ed Engl ; 61(13): e202101015, 2022 Mar 21.
Article in English | MEDLINE | ID: mdl-33590940

ABSTRACT

An adaptive coordination structure is vital for selective uranium extraction from seawater. By strategy of molecular imprinting, uranyl is introduced into a multivariate metal-organic framework (MOF) during the synthesis process to guide the in situ construction of proper nanocage structure for targeting uranyl binding. Except for the coordination between uranium with four oxygen from the materials, the axial oxygen of uranyl also forms hydrogen bonds with hydrogen from the phenolic hydroxyl group, which enhances the binding affinity of the material to uranyl. Attributing to the high binding affinity, the adsorbent shows high uranium binding selectivity to uranyl against not only the interfering metal ions, but also the carbonate group that coordinates with uranyl to form [UO2 (CO)3 ]4- in seawater. In natural seawater, the adsorbent realizes a high uranium adsorption capacity of 7.35 mg g-1 , together with an 18.38 times higher selectivity to vanadium.

5.
J Hazard Mater ; 424(Pt D): 127758, 2022 02 15.
Article in English | MEDLINE | ID: mdl-34801303

ABSTRACT

Uranium is a heavy metal with both chemotoxicity and radiotoxicity. Due to the increasing consumption of uranium, the remediation of uranium contamination and recovery of uranium from non-conventional approach is highly needed. Microorganism exhibits high potential for immobilization of uranium. This study for the first time isolated a marine Pseudomonas stutzeri strain MRU-UE1 with high uranium immobilization capacity of 308.72 mg/g, which is attributed to the synergetic mechanisms of biosorption, biomineralization, and bioreduction. The uranium is found to be immobilized in forms of tetragonal chernikovite (H2(UO2)2(PO4)2·8H2O) by biomineralization and CaU(PO4)2 by bioreduction under aerobic environment, which is rarely observed and would broaden the application of this strain in aerobic condition. The protein, phosphate group, and carboxyl group are found to be essential for the biosorption of uranium. In response to the stress of uranium, the strain produces inorganic phosphate group, which transformed soluble uranyl ion to insoluble uranium-containing precipitates, and poly-ß-hydroxybutyrate (PHB), which is observed for the first time during the interaction between microorganism and uranium. In summary, P. stutzeri strain MRU-UE1 would be a promising alternative for environmental uranium contamination remediation and uranium extraction from seawater.


Subject(s)
Pseudomonas stutzeri , Uranium , Biodegradation, Environmental , Biomineralization , Phosphates
6.
Res Vet Sci ; 139: 32-42, 2021 Oct.
Article in English | MEDLINE | ID: mdl-34246941

ABSTRACT

Porcine epidemic diarrhea virus (PEDV) is an enteropathogenic coronavirus; it causes diarrhea in pigs and is associated with high morbidity and mortality in sucking piglets. In this study, we performed in vitro and in vivo experiments to determine the inhibitory effects of Lactobacillus plantarum metabolites (LPM) on PEDV replication. Gas chromatography-mass spectrometry revealed exopolysaccharides to be one of the main components of LPM. We then determine whether L. plantarum exopolysaccharides (LPE) have an antiviral effect and also detected the expression levels of the apoptosis-related genes Bax and Bcl-2 and of the pro-apoptotic protein caspase-3. Further, we assessed the transcription levels of an immune-related protein (STAT1) and antiviral factors (MX1, MX2, ISG15, ZAP, PKR, and OAS1). Our results showed that the most effective method was to pretreat cells with LPM and that the optimal dose of LPM that could be safely administered to Vero cells was 1/8 times of the stock solution. LPE had a strong inhibitory effect on PEDV; the most effective method of administration was to co-incubate cells with LPE and PEDV, and the optimal concentration of LPE was 1.35 mg/mL. To conclude, LPE prevented PEDV adsorption and also alleviated inflammatory responses and induced early apoptosis of injured cells, but it could not regulate the immune function of cells.


Subject(s)
Lactobacillus plantarum/metabolism , Polysaccharides/pharmacology , Polysaccharides/therapeutic use , Porcine epidemic diarrhea virus/drug effects , Porcine epidemic diarrhea virus/growth & development , Swine Diseases/drug therapy , Swine Diseases/virology , Virus Replication/drug effects , Animals , Apoptosis/drug effects , Chlorocebus aethiops , Coronavirus Infections/drug therapy , Coronavirus Infections/immunology , Coronavirus Infections/veterinary , Coronavirus Infections/virology , Diarrhea/drug therapy , Diarrhea/veterinary , Diarrhea/virology , Inflammation/drug therapy , Porcine epidemic diarrhea virus/immunology , Swine , Swine Diseases/immunology , Vero Cells , Virus Attachment/drug effects
7.
Nat Commun ; 11(1): 5708, 2020 11 11.
Article in English | MEDLINE | ID: mdl-33177515

ABSTRACT

Extraction of uranium from seawater is critical for the sustainable development of nuclear energy. However, the currently available uranium adsorbents are hampered by co-existing metal ion interference. DNAzymes exhibit high selectivity to specific metal ions, yet there is no DNA-based adsorbent for extraction of soluble minerals from seawater. Herein, the uranyl-binding DNA strand from the DNAzyme is polymerized into DNA-based uranium extraction hydrogel (DNA-UEH) that exhibits a high uranium adsorption capacity of 6.06 mg g-1 with 18.95 times high selectivity for uranium against vanadium in natural seawater. The uranium is found to be bound by oxygen atoms from the phosphate groups and the carbonyl groups, which formed the specific nano-pocket that empowers DNA-UEH with high selectivity and high binding affinity. This study both provides an adsorbent for uranium extraction from seawater and broadens the application of DNA for being used in recovery of high-value soluble minerals from seawater.


Subject(s)
DNA, Catalytic/chemistry , DNA, Catalytic/metabolism , Seawater/chemistry , Uranium/isolation & purification , Chemical Fractionation/methods , Hydrogels , Oxygen/chemistry , Phosphates/chemistry , Photoelectron Spectroscopy , Uranium/metabolism
8.
Elife ; 92020 06 09.
Article in English | MEDLINE | ID: mdl-32513385

ABSTRACT

The kisspeptin system is a central modulator of the hypothalamic-pituitary-gonadal axis in vertebrates. Its existence outside the vertebrate lineage remains largely unknown. Here, we report the identification and characterization of the kisspeptin system in the sea cucumber Apostichopus japonicus. The gene encoding the kisspeptin precursor generates two mature neuropeptides, AjKiss1a and AjKiss1b. The receptors for these neuropeptides, AjKissR1 and AjKissR2, are strongly activated by synthetic A. japonicus and vertebrate kisspeptins, triggering a rapid intracellular mobilization of Ca2+, followed by receptor internalization. AjKissR1 and AjKissR2 share similar intracellular signaling pathways via Gαq/PLC/PKC/MAPK cascade, when activated by C-terminal decapeptide. The A. japonicus kisspeptin system functions in multiple tissues that are closely related to seasonal reproduction and metabolism. Overall, our findings uncover for the first time the existence and function of the kisspeptin system in a non-chordate species and provide new evidence to support the ancient origin of intracellular signaling and physiological functions that are mediated by this molecular system.


Subject(s)
Kisspeptins , Receptors, Kisspeptin-1 , Signal Transduction , Stichopus , Animals , Kisspeptins/genetics , Kisspeptins/metabolism , Kisspeptins/physiology , Receptors, Kisspeptin-1/genetics , Receptors, Kisspeptin-1/metabolism , Receptors, Kisspeptin-1/physiology , Signal Transduction/genetics , Signal Transduction/physiology , Stichopus/genetics , Stichopus/physiology
9.
Angew Chem Int Ed Engl ; 59(37): 15997-16001, 2020 09 07.
Article in English | MEDLINE | ID: mdl-32519451

ABSTRACT

The unique three-dimensional structure of spidrion determines the outstanding mechanical properties of the spider silk fiber. Inspired by the similarity of the three-dimensional structure of superb-uranyl binding protein (SUP) to that of spidroin, a dual-SUP (DSUP) chimeric protein fiber with high tensile strength is designed. The DSUP hydrogel fiber exhibits a loofah-shape structure by the cross-interaction of the protein nanofiber. Full exposure of abundant functional uranyl-binding sites in the stretchable loofah-shape hydrogel protein fiber give the DSUP fiber a groundbreaking uranium extraction capacity of 17.45 mg g-1 with an ultrashort saturation time of 3 days in natural seawater. This work reports the design of an adsorbent with ultrahigh uranium extraction capacity and explores a strategy for fabricating artificial high-strength functional non-spidroin protein fiber.

10.
Chem Commun (Camb) ; 56(28): 3935-3938, 2020 Apr 11.
Article in English | MEDLINE | ID: mdl-32196027

ABSTRACT

A bio-inspired cellulose paper-poly(amidoxime) composite hydrogel is explored via UV-polymerization. This hydrogel has a highly efficient uranium capture capacity of up to 6.21 mg g-1 for WU/Wdry gel and 12.9 mg g-1 for WU/Wpoly(amidoxime) in seawater for 6 weeks, due to its enhanced hydrophilicity, good hydraulic/ionic conductivity and broad-spectrum antibacterial performance.


Subject(s)
Anti-Bacterial Agents/chemistry , Cellulose/chemistry , Hydrogels/chemistry , Oximes/chemistry , Uranium/chemistry , Water Pollutants, Radioactive/chemistry , Water Purification/methods , Adsorption , Anti-Bacterial Agents/pharmacology , Cellulose/pharmacology , Escherichia coli/drug effects , Escherichia coli/growth & development , Hydrogels/pharmacology , Oximes/pharmacology , Paper , Seawater , Staphylococcus aureus/drug effects , Staphylococcus aureus/growth & development , Vibrio alginolyticus/drug effects , Vibrio alginolyticus/growth & development
11.
Virus Res ; 280: 197901, 2020 04 15.
Article in English | MEDLINE | ID: mdl-32070687

ABSTRACT

Transmissible gastroenteritis virus (TGEV) primarily replicates in intestinal epithelial cells and causes severe damage to host cells, resulting in diarrhea. Surface NHE3 serves as the key regulatory site controlling electroneutral Na+ absorption. In this study, our results showed that the surface NHE3 content was significantly reduced following TGEV infection, whereas the total level of protein expression was not significantly changed, and NHE3 activity gradually decreased with prolonged infection time. We then inhibited SGLT1 expression by lentiviral interference and drug inhibition, respectively. Inhibition studies showed that the level of phosphorylation of the downstream key proteins, MAPKAPK-2 and EZRIN, in the SGLT1-mediated p38MAPK/AKt2 signaling pathway was significantly increased. The surface NHE3 expression was also significantly increased, and NHE3 activity was also significantly enhanced. These results demonstrate that a TGEV infection can inhibit NHE3 translocation and attenuates sodium-hydrogen exchange activity via the SGLT1-mediated p38MAPK/AKt2 signaling pathway, affecting cellular electrolyte absorption leading to diarrhea.


Subject(s)
Enterocytes/virology , Proto-Oncogene Proteins c-akt/metabolism , Signal Transduction , Sodium-Glucose Transporter 1/genetics , Sodium-Hydrogen Exchanger 3/metabolism , p38 Mitogen-Activated Protein Kinases/metabolism , Animals , Cell Line , Proto-Oncogene Proteins c-akt/genetics , Sodium-Glucose Transporter 1/metabolism , Sodium-Hydrogen Exchanger 3/genetics , Swine , Transmissible gastroenteritis virus , p38 Mitogen-Activated Protein Kinases/genetics
12.
Angew Chem Int Ed Engl ; 59(11): 4262-4268, 2020 03 09.
Article in English | MEDLINE | ID: mdl-31908089

ABSTRACT

Biology has evolved excellent spatial structures for high-selectivity and high-affinity capture of heavy metals. Inspired by the spatial structure of the superb-uranyl binding protein SUP, we mimic the spatial structure of SUP in metal-organic frameworks (MOFs). The MOF UiO-66-3C4N fabricated by introducing 4-aminoisophthalic acid into UiO-66 shows high uranyl adsorption capacity both in simulated seawater and in natural seawater. In natural seawater, UiO-66-3C4N exhibits 17.03 times higher uranium extraction capacity than that of vanadium, indicating the high selectivity of the adsorbent. The EXAFS analysis and DFT calculation reveal that UiO-66-3C4N forms smaller nano-pocket for uranyl capture than that of SUP protein, which can both restrict the entrance of the other interfering ions with larger size and reinforce the binding by increasing the coordination interaction, and therefore qualify the nano-pocket with high affinity and high selectivity to uranyl.

13.
Angew Chem Int Ed Engl ; 59(3): 1220-1227, 2020 Jan 13.
Article in English | MEDLINE | ID: mdl-31692138

ABSTRACT

Based on the photoinduced photothermal, photoelectric, and photocatalytic effects of black phosphorus (BP) nanosheets, a BP-PAO fiber with enhanced uranium extraction capacity and high antibiofouling activity is fabricated by compositing BP nanosheets into polyacrylamidoxime (PAO). The photothermal effect increases the coordination interaction between UO2 2+ and the functional amidoxime group, and the photoelectric effect produces the surface positive electric field that exhibits electrostatic attraction to the negative [UO2 (CO3 )3 ]4- , which all increase the capacity for uranium adsorption. The photocatalytic effect endows the adsorbent with high antibiofouling activity by producing biotoxic reactive oxygen species. Owing to these three photoinduced effects, the photoinduced BP-PAO fiber shows a high uranium adsorption capacity of 11.76 mg g-1 , which is 1.50 times of the PAO fiber, in bacteria-containing natural seawater.

14.
Front Microbiol ; 10: 2540, 2019.
Article in English | MEDLINE | ID: mdl-31781061

ABSTRACT

Transmissible gastroenteritis (TGE), caused by transmissible gastroenteritis virus (TGEV), is one many gastrointestinal inflections in piglets, characterized by diarrhea, and high mortality. Probiotics are ubiquitous bacteria in animal intestines, which have many functions, such as promoting intestinal peristalsis and maintaining the intestinal balance. We found that the supernatant of the Lp-1 strain of Lactobacillus plantarum, isolated in our laboratory, and named Lp-1s had marked anti-TGEV effect on IPEC-J2 cells. Lp-1s could induce large amounts of interferon-ß in IPEC-J2 cells in the early stage (6 h) of infection with TGEV, and increased the level of phosphorylated signal transducer and activator of transcription and its nuclear translocation in the late stage (24-48 h) of infection. This resulted in upregulated expression of interferon-stimulated genes, and increased the transcription and protein expression of antiviral proteins, resulting in an anti-TGEV effect.

15.
Adv Sci (Weinh) ; 6(18): 1900961, 2019 Sep 18.
Article in English | MEDLINE | ID: mdl-31559134

ABSTRACT

Highly-efficient recovery of uranium from seawater is of great concern in the growing demand for nuclear energy. Bacteria are thought to be potential alternatives for uranium recovery. Herein, a Bacillus velezensis strain, UUS-1, with highly-efficient uranium immobilization capacity is isolated and is used in the recovery of uranium from seawater. The strain exhibits time-dependent uranium recovery capacity and only immobilizes uranium after growing for 12 h. The carboxyl group together with the amino group inside the bacterial cells, but not previously identified phosphate group, are essential for uranium immobilization. UUS-1 shows broad-spectrum antimicrobial activity by producing diverse antimicrobial metabolites, which endows the strain with innate resistance to the biofouling of marine microorganisms. Based on the dry weight of the initially used bacterial cultures, UUS-1 concentrates uranium by 6.26 × 105 times and reaches the high immobilization capacity of 9.46 ± 0.39 mg U g-1 bacterial cultures in real seawater within 48 h, which is the fastest uranium immobilization capacity observed from real seawater. Overall considering the ultrafast and highly-efficient uranium recovery capacity and the innate anti-biofouling activity, UUS-1 is a promising alternative for uranium recovery from seawater.

16.
Angew Chem Int Ed Engl ; 58(42): 14979-14985, 2019 Oct 14.
Article in English | MEDLINE | ID: mdl-31436013

ABSTRACT

By chemical cross-linking the amidoxime group onto dual-surfaces of natural ore materials, namely halloysite nanotubes (HNTs), an efficient adsorbent, AO-HNTs, is developed. AO-HNTs show high uranium adsorption capacity of 456.24 mg g-1 in 32 ppm uranium-spiked simulated seawater. In natural seawater, AO-HNTs reach the high uranium extraction capacity of 9.01 mg g-1 after 30 days' field test. The dual-surface amidoximated hollow nanotubular AO-HNTs exhibit more coordination active sites for uranium adsorption, which is attributed to the high and fast uranium adsorption capacity. Because of the stable natural ore structure, AO-HNTs also show long service life. Benefiting from the low cost of HNTs, the cost for uranium extraction from seawater is close to the uranium price in the spot uranium market, suggesting that AO-HNTs could be used for economical extraction of uranium from the oceans.

17.
Adv Sci (Weinh) ; 6(13): 1900002, 2019 Jul 03.
Article in English | MEDLINE | ID: mdl-31380181

ABSTRACT

The ocean reserves 4.5 billion tons of uranium and amounts to a nearly inexhaustible uranium supply. Biofouling in the ocean is one of the most severe factors that hazard uranium extraction and even cause the failure of uranium extraction. Therefore, development of uranium adsorbents with biofouling resistance is highly urgent. Herein, a strategy for constructing anti-biofouling adsorbents with enhanced uranium recovery capacity in natural seawater is developed. This strategy can be widely applied to modify currently available carboxyl-contained adsorbents, including the most popular amidoxime-based adsorbent and carboxyl metal organic framework adsorbent, using a simple one-step covalent cross-link reaction between the antibacterial compound and the adsorbent. The prepared anti-biofouling adsorbents display broad antibacterial spectrum and show more than 80% inhibition to the growth of marine bacteria. Benefitting from the tight covalent cross-link, the anti-biofouling adsorbents show high reusability. The modified amidoxime-based adsorbents show enhanced uranium recovery capacity both in sterilized and bacteria-contained simulated seawater. The anti-biofouling adsorbent Anti-UiO-66 constructed in this study exhibits 24.4% increased uranium recovery capacity, with a uranium recovery capacity of 4.62 mg-U per g-Ads, after a 30-day field test in real seawater, suggesting the strategy is a promising approach for constructing adsorbents with enhanced uranium extraction performance.

18.
Angew Chem Int Ed Engl ; 58(34): 11785-11790, 2019 08 19.
Article in English | MEDLINE | ID: mdl-31237392

ABSTRACT

For the practical extraction of uranium from seawater, adsorbents with high adsorption capacity, fast equilibrium rate, high selectivity, and long service life are needed. Herein, a chimeric spidroin-based super uranyl-binding protein (SSUP) fiber was designed by fusing the gene of super uranyl-binding protein (SUP) with the gene of spidroin. SUP endowed the SSUP fiber with high affinity and selectivity to uranium, and spidroin gave the SSUP fiber with high mechanical strength and high reusability. The wet SSUP fiber is a water-rich hydrogel-like structure, which provided abundant hydrophilic intermolecular space for the entrance of uranyl ions, and could accelerate the rate for uranium adsorption. In seawater, the SSUP fiber achieved a breakthrough uranium extraction capacity of 12.33 mg g-1 with an ultrashort equilibration time of 3.5 days, suggesting that SSUP fiber might be a promising adsorbent for uranium extraction from the natural seawater.

19.
Fish Shellfish Immunol ; 90: 297-307, 2019 Jul.
Article in English | MEDLINE | ID: mdl-31059811

ABSTRACT

Toll-like receptors (TLRs) play an important role in defense response to pathogens in mollusk. In this study the first TLR from Sepiella japonica (named as SjTLR) was functionally characterized, and its full-length cDNA consisted of 3914bp (GenBank accession no. AQY56780.1) including an open reading frame of 3582bp, encoding a putative protein of 1193 amino acids. Its theoretical molecular weight was 137.87 KDa and the predicted isoelectric point was 3.69. The derived amino acids sequence comprised of an extracellular domain including 26 amino acids signal peptide and eleven leucine-rich repeats (LRR), capped with LRRCT and LRRNT followed by transmembrane domain and cytoplasmic Toll/IL-1R domain (TIR). In addition, 12 potential N-linked glycosylation sites were present in the ectodomain to influence protein trafficking, surface presentation and ligand recognition. Multiple sequence alignment and phylogenetic analysis revealed that SjTLR shared the highest similarity to that of Euprymna scolopes and they fell into the same clade. Real-time PCR showed SjTLR expressed constitutively in all tested tissues, including gill, liver, brain, muscle, intestine, heart, lobus opticus and stomach, but showed different expression levels with genders. The highest expression was in the liver, and the lowest was in stomach for both genders. The functional domain region sequences encoding LRRs domain protein and TIR domain containing protein (TcpB) were expressed in BL21(DE3) respectively and purified with Ni-NAT Superflow resin conforming to the expected molecular weight. The cellular localization of SjTLR in HEK293 cells was conducted and plasma membrane localization was detected. SjLRRs internalization upon the activation of LPS was also observed, and dramatic redistribution of SjLRRs in the cytoplasm with distinct perinuclear accumulation was found. After SjTLR transfection Toll/NF-κB signaling pathway was active in HEK293 treated with LPS and TNFɑ. The nuclear related genes may also be activated by NF-κB in the nucleus, and the corresponding mRNA was transferred through the intracellular signal transduction pathway, so that IL-6 cytokines could be synthesized and released. After infection by Vibrio parahemolyticus and Aeromonas hydrophila the expression of SjTLR were upregulated with time-dependent manner. These findings might be valuable for understanding the innate immune signaling pathways of S.japonica and enabling future studies on host-pathogen interactions.


Subject(s)
Decapodiformes/genetics , Decapodiformes/immunology , Gene Expression Regulation/immunology , Immunity, Innate/genetics , Toll-Like Receptors/genetics , Toll-Like Receptors/immunology , Aeromonas hydrophila/physiology , Amino Acid Sequence , Animals , Base Sequence , Gene Expression Profiling , HEK293 Cells , Humans , Phylogeny , Sequence Alignment , Signal Transduction , Toll-Like Receptors/chemistry , Vibrio parahaemolyticus/physiology
20.
Vet Res ; 50(1): 28, 2019 Apr 27.
Article in English | MEDLINE | ID: mdl-31029162

ABSTRACT

Transmissible gastroenteritis coronavirus (TGEV) is an enteropathogenic coronavirus that causes diarrhea in pigs and is associated with high morbidity and mortality in sucking piglets. S1 is one of two protein domains in the spike (S) glycoprotein and is responsible for enteric tropism, sialic acid recognition, and host receptor binding. Although there has been extensive research on the S1 protein of TGEV, little is known about the intracellular role of TGEV-S1. In the present study, we used yeast two-hybrid screening of a cDNA library from porcine intestinal cells to identify proteins that interact with TGEV-S1. Among 120 positive clones from the library, 12 intracellular proteins were identified after sequencing and a BLAST search. These intracellular proteins are involved in protein synthesis and degradation, biological signal transduction, and negative control of signaling pathways. Using a glutathione-S-transferase (GST) pulldown assay and Co-IP, we found that UBXN1 interacts with the S1 protein. Here, we observed that TGEV infection led to increased UBXN1 expression levels during the late phase of infection in IPEC-J2 cells. Inhibition of UBXN1 in IPEC-J2 cells via siRNA interference significantly decreased the viral titer and downregulated the expression of S1. UBXN1 overexpression significantly increased the viral copy number. Additionally, we provided data suggesting that UBXN1 negatively regulates IFN-ß expression after TGEV infection. Finally, our research indicated that UBXN1 plays a vital role in the process of TGEV infection, making it a candidate target for the development of a novel antiviral method.


Subject(s)
Adaptor Proteins, Signal Transducing/physiology , Transmissible gastroenteritis virus/physiology , Viral Proteins/physiology , Virus Replication , Blotting, Western , Enzyme-Linked Immunosorbent Assay , Immunoprecipitation , RNA, Small Interfering/metabolism , Real-Time Polymerase Chain Reaction , Signal Transduction/physiology , Two-Hybrid System Techniques
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