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1.
EMBO Rep ; 25(3): 1055-1074, 2024 Mar.
Article in English | MEDLINE | ID: mdl-38351372

ABSTRACT

Activation of hepatic stellate cells (HSCs) plays a critical role in liver fibrosis. However, the molecular basis for HSC activation remains poorly understood. Herein, we demonstrate that primary cilia are present on quiescent HSCs but exhibit a significant loss upon HSC activation which correlates with decreased levels of the ciliary protein intraflagellar transport 88 (IFT88). Ift88-knockout mice are more susceptible to chronic carbon tetrachloride-induced liver fibrosis. Mechanistic studies show that the X-linked inhibitor of apoptosis (XIAP) functions as an E3 ubiquitin ligase for IFT88. Transforming growth factor-ß (TGF-ß), a profibrotic factor, enhances XIAP-mediated ubiquitination of IFT88, promoting its proteasomal degradation. Blocking XIAP-mediated IFT88 degradation ablates TGF-ß-induced HSC activation and liver fibrosis. These findings reveal a previously unrecognized role for ciliary homeostasis in regulating HSC activation and identify the XIAP-IFT88 axis as a potential therapeutic target for liver fibrosis.


Subject(s)
Cilia , Liver Cirrhosis , Animals , Mice , Cilia/metabolism , Hepatic Stellate Cells/metabolism , Hepatic Stellate Cells/pathology , Liver/metabolism , Liver Cirrhosis/chemically induced , Liver Cirrhosis/metabolism , Liver Cirrhosis/pathology , Transforming Growth Factor beta/metabolism
2.
Proc Natl Acad Sci U S A ; 120(2): e2208070120, 2023 01 10.
Article in English | MEDLINE | ID: mdl-36598944

ABSTRACT

Secretory proteins are cotranslationally or posttranslationally translocated across lipid membranes via a protein-conducting channel named SecY in prokaryotes and Sec61 in eukaryotes. The vast majority of secretory proteins in bacteria are driven through the channel posttranslationally by SecA, a highly conserved ATPase. How a polypeptide chain is moved by SecA through the SecY channel is poorly understood. Here, we report electron cryomicroscopy structures of the active SecA-SecY translocon with a polypeptide substrate. The substrate is captured in different translocation states when clamped by SecA with different nucleotides. Upon binding of an ATP analog, SecA undergoes global conformational changes to push the polypeptide substrate toward the channel in a way similar to how the RecA-like helicases translocate their nucleic acid substrates. The movements of the polypeptide substrates in the SecA-SecY translocon share a similar structural basis to those in the ribosome-SecY complex during cotranslational translocation.


Subject(s)
Bacterial Proteins , Escherichia coli Proteins , SecA Proteins/metabolism , Bacterial Proteins/metabolism , SEC Translocation Channels/metabolism , Models, Molecular , Protein Transport , Peptides/metabolism , Escherichia coli Proteins/metabolism
3.
Proc Natl Acad Sci U S A ; 120(19): e2220613120, 2023 05 09.
Article in English | MEDLINE | ID: mdl-37126722

ABSTRACT

Prostaglandin E2 (PGE2) and 16,16-dimethyl-PGE2 (dmPGE2) are important regulators of hematopoietic stem and progenitor cell (HSPC) fate and offer potential to enhance stem cell therapies [C. Cutler et al. Blood 122, 3074-3081(2013); W. Goessling et al. Cell Stem Cell 8, 445-458 (2011); W. Goessling et al. Cell 136, 1136-1147 (2009)]. Here, we report that PGE2-induced changes in chromatin at enhancer regions through histone-variant H2A.Z permit acute inflammatory gene induction to promote HSPC fate. We found that dmPGE2-inducible enhancers retain MNase-accessible, H2A.Z-variant nucleosomes permissive of CREB transcription factor (TF) binding. CREB binding to enhancer nucleosomes following dmPGE2 stimulation is concomitant with deposition of histone acetyltransferases p300 and Tip60 on chromatin. Subsequent H2A.Z acetylation improves chromatin accessibility at stimuli-responsive enhancers. Our findings support a model where histone-variant nucleosomes retained within inducible enhancers facilitate TF binding. Histone-variant acetylation by TF-associated nucleosome remodelers creates the accessible nucleosome landscape required for immediate enhancer activation and gene induction. Our work provides a mechanism through which inflammatory mediators, such as dmPGE2, lead to acute transcriptional changes and modify HSPC behavior to improve stem cell transplantation.


Subject(s)
Histones , Nucleosomes , Histones/metabolism , Chromatin , Dinoprostone , Regulatory Sequences, Nucleic Acid , Chromatin Assembly and Disassembly
4.
Plant J ; 119(3): 1336-1352, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38864745

ABSTRACT

Acacetin, a flavonoid compound, possesses a wide range of pharmacological effects, including antimicrobial, immune regulation, and anticancer effects. Some key steps in its biosynthetic pathway were largely unknown in flowering plants. Here, we present the first haplotype-resolved genome of Chrysanthemum indicum, whose dried flowers contain abundant flavonoids and have been utilized as traditional Chinese medicine. Various phylogenetic analyses revealed almost equal proportion of three tree topologies among three Chrysanthemum species (C. indicum, C. nankingense, and C. lavandulifolium), indicating that frequent gene flow among Chrysanthemum species or incomplete lineage sorting due to rapid speciation might contribute to conflict topologies. The expanded gene families in C. indicum were associated with oxidative functions. Through comprehensive candidate gene screening, we identified five flavonoid O-methyltransferase (FOMT) candidates, which were highly expressed in flowers and whose expressional levels were significantly correlated with the content of acacetin. Further experiments validated two FOMTs (CI02A009970 and CI03A006662) were capable of catalyzing the conversion of apigenin into acacetin, and these two genes are possibly responsible acacetin accumulation in disc florets and young leaves, respectively. Furthermore, combined analyses of ancestral chromosome reconstruction and phylogenetic trees revealed the distinct evolutionary fates of the two validated FOMT genes. Our study provides new insights into the biosynthetic pathway of flavonoid compounds in the Asteraceae family and offers a model for tracing the origin and evolutionary routes of single genes. These findings will facilitate in vitro biosynthetic production of flavonoid compounds through cellular and metabolic engineering and expedite molecular breeding of C. indicum cultivars.


Subject(s)
Chrysanthemum , Evolution, Molecular , Flavones , Genome, Plant , Phylogeny , Plant Proteins , Chrysanthemum/genetics , Chrysanthemum/metabolism , Chrysanthemum/enzymology , Flavones/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Genome, Plant/genetics , Haplotypes , Diploidy , Flavonoids/metabolism , Flavonoids/biosynthesis , Flowers/genetics , Flowers/enzymology , Flowers/metabolism , Methyltransferases/genetics , Methyltransferases/metabolism
5.
Gut ; 73(10): 1725-1736, 2024 Sep 09.
Article in English | MEDLINE | ID: mdl-38902029

ABSTRACT

OBJECTIVE: Hepatitis B surface antigen (HBsAg) loss is the optimal outcome for patients with chronic hepatitis B (CHB) but this rarely occurs with currently approved therapies. We aimed to develop and validate a prognostic model for HBsAg loss on treatment using longitudinal data from a large, prospectively followed, nationwide cohort. DESIGN: CHB patients receiving nucleos(t)ide analogues as antiviral treatment were enrolled from 50 centres in China. Quantitative HBsAg (qHBsAg) testing was prospectively performed biannually per protocol. Longitudinal discriminant analysis algorithm was used to estimate the incidence of HBsAg loss, by integrating clinical data of each patient collected during follow-up. RESULTS: In total, 6792 CHB patients who had initiated antiviral treatment 41.3 (IQR 7.6-107.6) months before enrolment and had median qHBsAg 2.9 (IQR 2.3-3.3) log10IU/mL at entry were analysed. With a median follow-up of 65.6 (IQR 51.5-84.7) months, the 5-year cumulative incidence of HBsAg loss was 2.4%. A prediction model integrating all qHBsAg values of each patient during follow-up, designated GOLDEN model, was developed and validated. The AUCs of GOLDEN model were 0.981 (95% CI 0.974 to 0.987) and 0.979 (95% CI 0.974 to 0.983) in the training and external validation sets, respectively, and were significantly better than those of a single qHBsAg measurement. GOLDEN model identified 8.5%-10.4% of patients with a high probability of HBsAg loss (5-year cumulative incidence: 17.0%-29.1%) and was able to exclude 89.6%-91.5% of patients whose incidence of HBsAg loss is 0. Moreover, the GOLDEN model consistently showed excellent performance among various subgroups. CONCLUSION: The novel GOLDEN model, based on longitudinal qHBsAg data, accurately predicts HBsAg clearance, provides reliable estimates of functional hepatitis B virus (HBV) cure and may have the potential to stratify different subsets of patients for novel anti-HBV therapies.


Subject(s)
Antiviral Agents , Hepatitis B Surface Antigens , Hepatitis B, Chronic , Humans , Hepatitis B Surface Antigens/blood , Hepatitis B, Chronic/drug therapy , Hepatitis B, Chronic/blood , Antiviral Agents/therapeutic use , Male , Female , Middle Aged , Adult , Prospective Studies , China/epidemiology , Longitudinal Studies , Hepatitis B virus/immunology , Prognosis
6.
J Cell Physiol ; 239(5): e31215, 2024 May.
Article in English | MEDLINE | ID: mdl-38308657

ABSTRACT

Primary cilia are distributed extensively within the corneal epithelium and endothelium. However, the presence of cilia in the corneal stroma and the dynamic changes and roles of endothelial and stromal cilia in corneal homeostasis remain largely unknown. Here, we present compelling evidence for the presence of primary cilia in the corneal stroma, both in vivo and in vitro. We also demonstrate dynamic changes of both endothelial and stromal cilia during corneal development. In addition, our data show that cryoinjury triggers dramatic cilium formation in the corneal endothelium and stroma. Furthermore, depletion of cilia in mutant mice lacking intraflagellar transport protein 88 compromises the corneal endothelial capacity to establish the effective tissue barrier, leading to an upregulation of α-smooth muscle actin within the corneal stroma in response to cryoinjury. These observations underscore the essential involvement of corneal endothelial and stromal cilia in maintaining corneal homeostasis and provide an innovative strategy for the treatment of corneal injuries and diseases.


Subject(s)
Cilia , Corneal Stroma , Endothelium, Corneal , Homeostasis , Animals , Mice , Actins/metabolism , Cilia/metabolism , Corneal Injuries/metabolism , Corneal Injuries/pathology , Corneal Injuries/therapy , Corneal Stroma/cytology , Corneal Stroma/growth & development , Corneal Stroma/metabolism , Endothelium, Corneal/cytology , Endothelium, Corneal/growth & development , Endothelium, Corneal/metabolism , Homeostasis/physiology , Mice, Inbred C57BL , Mice, Knockout , Tumor Suppressor Proteins/genetics , Ciliopathies/metabolism , Ciliopathies/pathology , Ciliopathies/therapy
7.
Article in English | MEDLINE | ID: mdl-39181427

ABSTRACT

BACKGROUND AND AIMS: Hepatocellular carcinoma (HCC) risk prediction models established in patients with chronic hepatitis B receiving a nucleos(t)ide analogue (NA) rarely include viral factors because of mediocre predictability of traditional viral markers. Here, we investigate the role of serum hepatitis B virus (HBV) RNA, a novel biomarker, in predicting HCC risk in NA-treated patients. METHODS: A total of 1374 NA-treated patients were enrolled from 2 prospective chronic hepatitis B cohorts. Serum HBV RNA was detected at baseline, year 1, 2 and 3 of treatment. Cox proportional-hazard model was used to investigate the association of HBV RNA kinetics with HCC risk. RESULTS: After a median follow-up of 5.4 years, 76 patients developed HCC. HBV RNA declines at year 1 (adjusted hazard ratio, 0.70, P = .009) and 2 (adjusted hazard ratio, 0.71; P = .016) were independently associated with HCC risk. Patients with less HBV RNA decline at year 1 (≤0.4 log10 copies/mL) or 2 (≤0.6 log10 copies/mL) had 2.22- and 2.09-folds higher HCC risk, respectively, than those with more declines. When incorporating these early on-treatment HBV RNA declines into existing HCC risk scores, including PAGE-B (age, sex, and platelets), modified PAGE-B (mPAGE-B) (age, sex, platelets, and albumin), and aMAP (age, sex, platelets, and albumin-bilirubin score) score, they could enhance their predictive performance (ie, C-index 0.814 vs 0.78 [model (PAGE-B + year-1 HBV RNA decline) vs PAGE-B score based on baseline parameters]). CONCLUSIONS: Serum HBV RNA declines at year 1 and 2 were significantly associated with on-treatment HCC risk. Incorporating early on-treatment HBV RNA declines into HCC risk prediction models can be useful tools to guide appropriate surveillance strategies in NA-treated patients.

8.
J Med Virol ; 96(4): e29510, 2024 Apr.
Article in English | MEDLINE | ID: mdl-38573018

ABSTRACT

Hepatitis B virus (HBV) infection poses a significant burden on global public health. Unfortunately, current treatments cannot fully alleviate this burden as they have limited effect on the transcriptional activity of the tenacious covalently closed circular DNA (cccDNA) responsible for viral persistence. Consequently, the HBV life cycle should be further investigated to develop new anti-HBV pharmaceutical targets. Our previous study discovered that the host gene TMEM203 hinders HBV replication by participating in calcium ion regulation. The involvement of intracellular calcium in HBV replication has also been confirmed. In this study, we found that transient receptor potential vanilloid 4 (TRPV4) notably enhances HBV reproduction by investigating the effects of several calcium ion-related molecules on HBV replication. The in-depth study showed that TRPV4 promotes hepatitis B core/capsid protein (HBc) protein stability through the ubiquitination pathway and then promotes the nucleocapsid assembly. HBc binds to cccDNA and reduces the nucleosome spacing of the cccDNA-histones complex, which may regulate HBV transcription by altering the nucleosome arrangement of the HBV genome. Moreover, our results showed that TRPV4 promotes cccDNA-dependent transcription by accelerating the methylation modification of H3K4. In conclusion, TRPV4 could interact with HBV core protein and regulate HBV during transcription and replication. These data suggest that TRPV4 exerts multifaceted HBV-related synergistic factors and may serve as a therapeutic target for CHB.


Subject(s)
Antineoplastic Agents , Hepatitis B , Humans , Ubiquitin , Capsid , Capsid Proteins , Hepatitis B virus/genetics , TRPV Cation Channels/genetics , Calcium , Nucleosomes , Methylation , Membrane Proteins
9.
Plant Physiol ; 192(3): 2419-2435, 2023 07 03.
Article in English | MEDLINE | ID: mdl-36932696

ABSTRACT

Terpene synthases (TPSs) play pivotal roles in conferring the structural diversity of terpenoids, which are mainly emitted from flowers, whereas the genetic basis of the release of floral volatile terpenes remains largely elusive. Though quite similar in sequence, TPS allelic variants still function divergently, and how they drive floral terpene diversity in closely related species remains unknown. Here, TPSs responsible for the floral scent of wild Freesia species were characterized, and the functions of their natural allelic variants, as well as the causal amino acid residues, were investigated in depth. Besides the 8 TPSs previously reported in modern cultivars, 7 additional TPSs were functionally evaluated to contribute to the major volatiles emitted from wild Freesia species. Functional characterization of allelic natural variants demonstrated that allelic TPS2 and TPS10 variants changed the enzymatic capacity while allelic TPS6 variants drove the diversity of floral terpene products. Further residue substitution analysis revealed the minor residues determining the enzyme catalytic activity and product specificity. The clarification of TPSs in wild Freesia species reveals that allelic TPS variants evolved differently to determine the interspecific floral volatile terpenes in the genus and might be used for modern cultivar improvement.


Subject(s)
Alkyl and Aryl Transferases , Terpenes , Terpenes/metabolism , Phylogeny , Alkyl and Aryl Transferases/genetics
10.
Opt Express ; 32(7): 11895-11906, 2024 Mar 25.
Article in English | MEDLINE | ID: mdl-38571027

ABSTRACT

Pure-quartic solitons (PQSs) have recently received increasing attention due to their energy-width scaling over the traditional soliton, which has expanded our understanding of soliton dynamics with high-order dispersion in nonlinear systems. Here, we numerically reveal the asynchronization and synchronization processes of the sub-pulse within the vector PQS molecule in a mode-locked fiber laser by solving the coupled Ginzburg-Landau equations. During the establishment of a vector PQS molecule, the repulsion, attraction, and finally stabilization processes have been observed. Specifically, sub-pulse disappearance, regeneration, and finally synchronization with the other pulses are also investigated. Our analysis of the pulse energy, time interval, and relative phase evolution dynamics with the round trip indicates that the asynchronization and synchronization within the vector PQS molecule associate tightly with the gain competition and the cross-phase modulation. Our findings provide insights into the internal mutual dynamics within the vector soliton molecule and offer guidance for the applications of PQS.

11.
FASEB J ; 37(7): e23035, 2023 07.
Article in English | MEDLINE | ID: mdl-37310396

ABSTRACT

Nonalcoholic steatohepatitis (NASH) has become a major concern that threatens human health worldwide. The underlying pathogenesis was crucial but remained poorly understood. Here, we found that the expression of hepatic farnesyl diphosphate synthase (FDPS) was increased in mice and patients with NASH. Elevated FDPS levels were positively correlated with NASH severity. Overexpression of FDPS in mice provoked increased lipid accumulation, inflammation, and fibrosis, while hepatic FDPS deficiency protected mice from NASH progression. Importantly, pharmacological inhibition of FDPS with clinically used alendronate remarkably attenuated NASH-associated phenotypes in mice. Mechanistically, we demonstrated that FDPS increased its downstream product farnesyl pyrophosphate levels, which could function as an aryl hydrocarbon receptor (AHR) agonist to upregulate the expression of fatty acid translocase CD36, to accelerate the development of NASH. Collectively, these findings suggest that FDPS exacerbates NASH via AHR-CD36 axis and identify FDPS as a promising target for NASH therapy.


Subject(s)
Non-alcoholic Fatty Liver Disease , Animals , Humans , Mice , Alendronate , CD36 Antigens/genetics , Geranyltranstransferase/genetics , Receptors, Aryl Hydrocarbon/genetics
12.
Langmuir ; 40(1): 906-914, 2024 Jan 09.
Article in English | MEDLINE | ID: mdl-38130111

ABSTRACT

The unique two-dimensional layered structure of BiOCl makes it highly promising for energy storage applications. In this study, we successfully synthesized BiOCl nanoparticles encapsulated in N-doped carbon nanonecklaces (BiOCl NPs/N-CNNs) using well-established electrospinning and solvothermal substitution. As an anode material for lithium-ion batteries, BiOCl NPs/N-CNNs exhibited enhanced rate performance, delivering a capacity of 220.2 mA h g-1 at 8 A g-1. Furthermore, it demonstrated remarkable long cycle stability, retaining a capacity of 200.5 mA h g-1 after 9000 cycles with a discharge rate of 8.0 A g-1. The superior electrochemical performance can be attributed to the stacked layered structure of BiOCl, facilitated by van der Waals force, as well as the ingenious nanonecklace structures. These structures not only provide fast ion diffusion pathways but also enhance electrolyte penetration and offer more active sites for Li+ insertion and extraction. Additionally, the nanonecklace structure prevents the aggregation of nanopolyhedra, promoting the complete reaction of BiOCl with Li+. Moreover, the unique nanopolyhedron structure alleviates the stress caused by the volume expansion of Bi nanoparticles during cycling and reduces the internal resistance of the electrode.

13.
Horm Metab Res ; 2024 Oct 14.
Article in English | MEDLINE | ID: mdl-39401523

ABSTRACT

Gestational diabetes mellitus (GDM) is a common metabolic disorder in pregnancy and leads to serious harm to the mother and the fetus. A variety of lncRNAs play a key role in GDM. This meta-analysis was performed to explore the potential value of lncRNAs in GDM diagnosis. Articles correlated with lncRNA and GDM were screened from Embase, Medline, EBSCO, PubMed, Chinese National Knowledge Infrastructure, and WanFang databases. Summary receiver operator characteristic (SROC) was performed to evaluate the pooled area under curve (AUC). Forest plot was conducted to calculate the sensitivity, specificity, diagnostic likelihood ratio (LR), diagnostic score, and diagnostic odds ratio (DOR). Deeks' funnel plot was utilized to evaluate the publication bias. Eleven articles containing 12 tests (1060 GDM patients and 1066 controls) were included in this meta-analysis. AUC (0.89, 95%CI=0.86-0.92), sensitivity (0.84, 95%CI=0.80-0.87), and specificity (0.81, 95%CI=0.77-0.85)of the SROC curve showed a high diagnostic value of lncRNA for GDM. Positive LR (PLR 4.40, 95%CI=3.45-5.60) and negative LR (NLR 0.20, 95%CI=0.15-0.26) results indicated that the diagnosis of lncRNA for GDM had low clinical utility. Diagnostic score (3.09, 95%CI=2.62-3.57) and DOR (22.04, 95%CI=13.68-35.51) results suggested lncRNAs have good discriminative effect on GDM. Heterogeneity was significantly higher, but not induced by the subgroups. LncRNAs had high diagnostic value and good discriminative effect for GDM, but the clinical utility was not high. This meta-analysis study offers a potential target for GDM diagnosis.

14.
World J Urol ; 42(1): 238, 2024 Apr 16.
Article in English | MEDLINE | ID: mdl-38627315

ABSTRACT

BACKGROUND: Accurate estimation of the glomerular filtration rate (GFR) is clinically crucial for determining the status of obstruction, developing treatment strategies, and predicting prognosis in obstructive nephropathy (ON). We aimed to develop a deep learning-based system, named UroAngel, for non-invasive and convenient prediction of single-kidney function level. METHODS: We retrospectively collected computed tomography urography (CTU) images and emission computed tomography diagnostic reports of 520 ON patients. A 3D U-Net model was used to segment the renal parenchyma, and a logistic regression multi-classification model was used to predict renal function level. We compared the predictive performance of UroAngel with the Modification of Diet in Renal Disease (MDRD), Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equations, and two expert radiologists in an additional 40 ON patients to validate clinical effectiveness. RESULTS: UroAngel based on 3D U-Net convolutional neural network could segment the renal cortex accurately, with a Dice similarity coefficient of 0.861. Using the segmented renal cortex to predict renal function stage had high performance with an accuracy of 0.918, outperforming MDRD and CKD-EPI and two radiologists. CONCLUSIONS: We proposed an automated 3D U-Net-based analysis system for direct prediction of single-kidney function stage from CTU images. UroAngel could accurately predict single-kidney function in ON patients, providing a novel, reliable, convenient, and non-invasive method.


Subject(s)
Deep Learning , Renal Insufficiency, Chronic , Solitary Kidney , Humans , Retrospective Studies , Kidney/diagnostic imaging , Renal Insufficiency, Chronic/diagnosis , Glomerular Filtration Rate , Tomography , Creatinine
15.
Stat Med ; 43(10): 1920-1932, 2024 May 10.
Article in English | MEDLINE | ID: mdl-38417455

ABSTRACT

Consider the choice of outcome for overall treatment benefit in a clinical trial which measures the first time to each of several clinical events. We describe several new variants of the win ratio that incorporate the time spent in each clinical state over the common follow-up, where clinical state means the worst clinical event that has occurred by that time. One version allows restriction so that death during follow-up is most important, while time spent in other clinical states is still accounted for. Three other variants are described; one is based on the average pairwise win time, one creates a continuous outcome for each participant based on expected win times against a reference distribution and another that uses the estimated distributions of clinical state to compare the treatment arms. Finally, a combination testing approach is described to give robust power for detecting treatment benefit across a broad range of alternatives. These new methods are designed to be closer to the overall treatment benefit/harm from a patient's perspective, compared to the ordinary win ratio. The new methods are compared to the composite event approach and the ordinary win ratio. Simulations show that when overall treatment benefit on death is substantial, the variants based on either the participants' expected win times (EWTs) against a reference distribution or estimated clinical state distributions have substantially higher power than either the pairwise comparison or composite event methods. The methods are illustrated by re-analysis of the trial heart failure: a controlled trial investigating outcomes of exercise training.


Subject(s)
Heart Failure , Humans , Endpoint Determination/methods , Data Interpretation, Statistical
16.
Microb Cell Fact ; 23(1): 5, 2024 Jan 03.
Article in English | MEDLINE | ID: mdl-38172868

ABSTRACT

BACKGROUND AND AIM: Reprogramming microorganisms to enhance the production of metabolites is a part of contemporary synthetic biology, which relies on the availability of genetic tools to successfully manipulate the bacteria. Methylorubrum extorquens AM1 is a platform microorganism used to convert C1 compounds into various value-added products. However, the repertoire of available plasmids to conveniently and quickly fine-tune the expression of multiple genes in this strain is extremely limited compared with other model microorganisms such as Escherichia coli. Thus, this study aimed to integrate existing technologies, such as transposon-mediated chromosomal integration and cre-lox-mediated recombination, to achieve the diversified expression of target genes through multiple chromosomal insertions in M. extorquens AM1. RESULTS: A single plasmid toolkit, pSL-TP-cre-km, containing a miniHimar1 transposon and an inducible cre-lox71/lox66 system, was constructed and characterized for its multiple chromosomal integration capacity. A co-transcribed mcr-egfp cassette [for the production of 3-hydroxypropionic acid (3-HP) and a reporting green fluorescent protein] was added to construct pTP-cre-mcr-egfp for evaluating its utility in mediating the expression of heterologous genes, resulting in the production of 3-HP with a titer of 34.7-55.2 mg/L by two chromosomal integration copies. Furthermore, in association with the expression of plasmid-based mcr, 3-HP production increased to 65.5-92.4 mg/L. CONCLUSIONS: This study used a multi-round chromosomal integration system based on cre-lox71/lox66 and a transposon to construct a single constructed vector. A heterologous mcr gene was introduced through this vector, and high expression of 3-hydroxypropionic acid was achieved in M. extorquens. This study provided an efficient genetic tool for manipulating M. extorquens, which not only help increase the expression of heterologous genes in M. extorquens but also provide a reference for strains lacking genetic manipulation vectors.


Subject(s)
Genetic Vectors , Integrases , Integrases/genetics , Plasmids/genetics , Lactic Acid
17.
J Org Chem ; 89(6): 3987-3994, 2024 Mar 15.
Article in English | MEDLINE | ID: mdl-38437716

ABSTRACT

A novel catalyst-free cascade amination/cyclization/reduction reaction was developed for the synthesis of various Dihydroquinoxalinones under mild conditions from accessible biomass-derived keto acids and 1,2-phenylenediamines with ammonia borane as a hydrogen donor. This single-step approach enables a simple and eco-friendly route toward the direct synthesis of 12 kinds of Dihydroquinoxalinones in moderate to excellent yields in the green solvent dimethyl carbonate. The results of deuterium-labeling experiments and density function calculations demonstrate that the reductive process proceeds along a double hydrogen transfer pathway. An acceptable yield of Dihydroquinoxalinone can be afforded in a gram-scale experiment, illustrating the practicality of the as-reported reaction system.

18.
Fish Shellfish Immunol ; 146: 109418, 2024 Mar.
Article in English | MEDLINE | ID: mdl-38301811

ABSTRACT

The rice fish system represents an innovative and sustainable approach to integrated farming, combining rice cultivation with fish rearing in the same ecosystem. However, one of the major challenges in this system is the pesticidal pollution resulting from various sources, which poses risks to fish health and overall ecosystem balance. In recent years, dietary polyphenols have emerged as promising bioactive compounds with potential chemo-preventive and therapeutic properties. These polyphenols, derived from various plant sources, have shown great potential in reducing the toxicity of pesticides and improving the health of fish within the rice fish system. This review aims to explore the novel aspects of using dietary polyphenols to mitigate pesticidal toxicity and enhance fish health in the rice fish system. It provides comprehensive insights into the mechanisms of action of dietary polyphenols and their beneficial effects on fish health, including antioxidant, anti-inflammatory, and detoxification properties. Furthermore, the review discusses the potential application methods of dietary polyphenols, such as direct supplementation in fish diets or through incorporation into the rice fields. By understanding the interplay between dietary polyphenols and pesticides in the rice fish system, researchers can develop innovative and sustainable strategies to promote fish health, minimize pesticide impacts, and ensure the long-term viability of this integrated farming approach. The information presented in this review will be valuable for scientists, aqua-culturists, and policymakers aiming to implement eco-friendly and health-enhancing practices in the rice fish system.


Subject(s)
Oryza , Pesticides , Animals , Ecosystem , Agriculture/methods , Pesticides/toxicity , Pesticides/analysis , Fishes , Polyphenols/pharmacology
19.
Inorg Chem ; 63(29): 13253-13264, 2024 Jul 22.
Article in English | MEDLINE | ID: mdl-38984385

ABSTRACT

Efficient CO oxidation at ambient or low temperatures is essential for environmental purification and selective CO oxidation in H2, yet achieving this remains a challenge with current methodologies. In this research, we extensively evaluated the catalytic performance of phosphotungstic acid (PTA)-supported 11 M1/PTA single-atom catalysts (SACs) using density functional theory calculations across both gas phase and 12 common solvents. The Rh1/PTA, Pd1/PTA, and Pt1/PTA systems exhibit moderate CO adsorption energies, facilitating the feasibility of oxygen vacancy formation. Remarkably, the Pd1/PTA and Pt1/PTA catalysts exhibited negligible energy barriers and demonstrated exceptionally high catalytic rates, with values reaching up to (1 × 1010)11, markedly exceeding the threshold for room temperature reactions, set at 6.55 × 108. This phenomenon is attributed to a transition from the high-energy barrier processes of oxygen dissociation in O2 and N-O bond dissociation in N2O to the more efficient dissociation of H2O2. Orbital analysis and charge variations at metal sites throughout the reaction process provide deeper insights into the role of the three metal catalytic sites in CO activation. Our findings not only reveal key aspects of SACs in facilitating CO oxidation at low temperatures but also provide valuable insights for future catalytic reaction mechanism studies and environmental applications.

20.
Fish Shellfish Immunol ; 154: 109918, 2024 Sep 21.
Article in English | MEDLINE | ID: mdl-39307257

ABSTRACT

Gill remodeling is an important strategy for fish to cope with hypoxia, and many of the teleost possess this ability, but the underlying mechanism is not well understood. To investigate the mechanism of hypoxia-induced gill remodeling, largemouth bass (Micropterus salmoides) exposed to hypoxia (dissolved oxygen level: 2.0 ± 0.2 mg L-1) for 7 days, followed by 7 days of reoxygenation. Hypoxia tests were also performed on primary gill cells from largemouth bass. We found that hypoxia-induced gill remodeling increased the respiratory surface area of the gills. This change in gill morphology was reversible and recovered after reoxygenation. A reduction in the number of mucous cells and rearrangement of mitochondria-rich cells (MRCs) were observed during gill remodeling. After 7 days of reoxygenation, the number of mucous cells and the position of the MRCs were restored. Hypoxia resulted in a 2.92-fold increase in the number of primary gill cells that underwent migration over a 12-h period. The mRNA levels of nine integrin subunits (α1, α2, α5, α7, α8, α10, αL, ß1 and ß2) were significantly up-regulated after 12 h of hypoxia in vivo, and the changes in the expression of these subunits were consistent with the HIF-1α trend. Immunohistochemistry showed that integrin ß1 protein levels were significantly increased and were abundantly expressed in the interlamellar cell mass after exposure to hypoxia. Taken together, the results of the present study demonstrated that changes in mucosal cells and MRCs play an important role in hypoxia-induced gill remodeling in largemouth bass and that these changes are regulated by integrins.

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