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Pesticide Biochemistry and Physiology Jun 2024Lufenuron, a benzoylurea chitin synthesis inhibitor, is effective against many insect pests. However, the insecticidal activity of lufenuron has not been completely...
Lufenuron, a benzoylurea chitin synthesis inhibitor, is effective against many insect pests. However, the insecticidal activity of lufenuron has not been completely elucidated, nor has its disturbing effect on chitin synthesis genes. In this study, bioassay results demonstrated an outstanding toxicity of lufenuron against Helicoverpa armigera larvae. The treated larvae died from abortive molting and metamorphosis defects, and severe separation of epidermis and subcutaneous tissues was observed. Treatment of 3rd- and 4th-instar larvae with LC lufenuron significantly extended the duration of larval and pupal stage, reduced the rates of pupation and emergence, and adversely affected pupal weight. Besides, lufenuron can severely reduce chitin content in larval integument, and the lufenuron-treated larvae showed reduced trehalose content in their hemolymph. Further analysis using RNA sequencing revealed that five chitin synthesis genes were down-regulated, whereas the expressions of two chitin degradation genes were significantly enhanced. Knockdown of chitin synthase 1 (HaCHS1), uridine diphosphate-N-acetylglucosamine-pyrophosphorylase (HaUAP), phosphoacetyl glucosamine mutase (HaPGM), and glucosamine 6-phosphate N-acetyl-transferase (HaGNPAT) in H. armigera led to significant increase in larval susceptibilities to LC lufenuron by 75.48%, 65.00%, 68.42% and 28.00%, respectively. Our findings therefore revealed the adverse effects of sublethal doses of lufenuron on the development of H. armigera larvae, elucidated the perturbations on chitin metabolism, and proved that the combination of RNAi and lufenuron would improve the control effect of this pest.
Topics: Animals; Chitin; Benzamides; Larva; Insecticides; Moths; Insect Proteins; Chitin Synthase; Helicoverpa armigera; Fluorocarbons
PubMed: 38879310
DOI: 10.1016/j.pestbp.2024.105962 -
Journal of Agricultural and Food... Jun 20243-Fucosyllactose (3-FL), an important fucosylated human milk oligosaccharide in breast milk, offers numerous health benefits to infants. Previously, we metabolically...
3-Fucosyllactose (3-FL), an important fucosylated human milk oligosaccharide in breast milk, offers numerous health benefits to infants. Previously, we metabolically engineered BL21(DE3) for the biosynthesis of 3-FL. In this study, we initially optimized culture conditions to double 3-FL production. Competing pathway genes involved in guanosine 5'-diphosphate-fucose biosynthesis were subsequently inactivated to redirect fluxes toward 3-FL biosynthesis. Next, three promising transporters were evaluated using plasmid-based or chromosomally integrated expression to maximize extracellular 3-FL production. Additionally, through analysis of α1,3-fucosyltransferase (FutM2) structure, we identified Q126 residues as a highly mutable residue in the active site. After site-saturation mutation, the best-performing mutant, FutM2-Q126A, was obtained. Structural analysis and molecular dynamics simulations revealed that small residue replacement positively influenced helical structure generation. Finally, the best strain BD3-A produced 6.91 and 52.1 g/L of 3-FL in a shake-flask and fed-batch cultivations, respectively, highlighting its potential for large-scale industrial applications.
Topics: Escherichia coli; Trisaccharides; Fucosyltransferases; Metabolic Engineering; Humans; Oligosaccharides
PubMed: 38878091
DOI: 10.1021/acs.jafc.4c02950 -
The Journal of Physical Chemistry. B Jun 2024The G protein-coupled receptors (GPCRs) play a pivotal role in numerous biological processes as crucial cell membrane receptors. However, the dynamic mechanisms...
The G protein-coupled receptors (GPCRs) play a pivotal role in numerous biological processes as crucial cell membrane receptors. However, the dynamic mechanisms underlying the activation of GPR183, a specific GPCR, remain largely elusive. To address this, we employed computational simulation techniques to elucidate the activation process and key events associated with GPR183, including conformational changes from inactive to active state, binding interactions with the G protein complex, and GDP release. Our findings demonstrate that the association between GPR183 and the G protein involves the formation of receptor-specific conformations, the gradual proximity of the G protein to the binding pocket, and fine adjustments of the protein conformation, ultimately leading to a stable GPR183-G complex characterized by a high energy barrier. The presence of G protein partially promotes GPR183 activation, which is consistent with the observation of GPCR constitutive activity test experiments, thus illustrating the reliability of our calculations. Moreover, our study suggests the existence of a stable partially activated state preceding complete activation, providing novel avenues for future investigations. In addition, the relevance of GPR183 for various diseases, such as colitis, the response of eosinophils to infection, antiviral properties, and pulmonary inflammation, has been emphasized, underscoring its therapeutic potential. Consequently, understanding the activation process of GPR183 through molecular dynamic simulations offers valuable kinetic insights that can aid in the development of targeted therapies.
Topics: Receptors, G-Protein-Coupled; Molecular Dynamics Simulation; Humans; Protein Conformation; Guanosine Diphosphate; Protein Binding; Binding Sites; GTP-Binding Protein alpha Subunits, Gi-Go
PubMed: 38877985
DOI: 10.1021/acs.jpcb.4c02812 -
Clinical and Translational Science Jun 2024This cohort study aims to assess the connection between cytochrome P450 family 2 subfamily C member 19 (CYP2C19) genotyping, platelet aggregability following oral...
This cohort study aims to assess the connection between cytochrome P450 family 2 subfamily C member 19 (CYP2C19) genotyping, platelet aggregability following oral clopidogrel administration, and the occurrence of postoperative atrial fibrillation (POAF) after off-pump coronary artery bypass graft (CABG) surgery. From May 2017 to November 2022, a total of 258 patients undergoing elective first-time CABG surgery, receiving 100 mg/day oral aspirin and 75 mg/day oral clopidogrel postoperatively, was included for analysis. These patients were categorized based on CYP2C19 genotyping. Platelet aggregability was assessed serially using multiple-electrode aggregometry before CABG, 1 and 5 days after the procedure, and before discharge. The incidences of POAF were compared using the log-rank test for cumulative risk. CYP2C19 genotyping led to categorization into CYP2C19*1*1 (WT group, n = 123) and CYP2C19*2 or *3 (LOF group, n = 135). Baseline characteristics and operative data showed no significant differences between the two groups. The incidence of POAF after CABG was 42.2% in the LOF group, contrasting with 22.8% in the WT group (hazard risk [HR]: 2.061; 95% confidence interval [CI]: 1.347, 3.153; p = 0.0013). Adenosine diphosphate-stimulated platelet aggregation was notably higher in the LOF group compared to the WT group 5 days after CABG (30.4% ± 6.5% vs. 17.9% ± 4.1%, p < 0.001), remaining a similar higher level at hospital discharge (25.6% ± 6.1% vs. 12.2% ± 3.5%, p < 0.001). The presence of CYP2C19 LOF was linked to a higher incidence of POAF and relatively elevated platelet aggregation after CABG surgery under the same oral clopidogrel regimen.
Topics: Humans; Cytochrome P-450 CYP2C19; Atrial Fibrillation; Male; Female; Aged; Coronary Artery Bypass; Middle Aged; Clopidogrel; Postoperative Complications; Genotype; Platelet Aggregation Inhibitors; Platelet Aggregation; Incidence; Aspirin
PubMed: 38877696
DOI: 10.1111/cts.13862 -
European Urology Focus Jun 2024Defining optimal therapeutic sequencing strategies in prostate cancer (PC) is challenging and may be assisted by artificial intelligence (AI)-based tools for an analysis...
BACKGROUND
Defining optimal therapeutic sequencing strategies in prostate cancer (PC) is challenging and may be assisted by artificial intelligence (AI)-based tools for an analysis of the medical literature.
OBJECTIVE
To demonstrate that INSIDE PC can help clinicians query the literature on therapeutic sequencing in PC and to develop previously unestablished practices for evaluating the outputs of AI-based support platforms.
DESIGN, SETTING, AND PARTICIPANTS
INSIDE PC was developed by customizing PubMed Bidirectional Encoder Representations from Transformers. Publications were ranked and aggregated for relevance using data visualization and analytics. Publications returned by INSIDE PC and PubMed were given normalized discounted cumulative gain (nDCG) scores by PC experts reflecting ranking and relevance.
INTERVENTION
INSIDE PC for AI-based semantic literature analysis.
OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS
INSIDE PC was evaluated for relevance and accuracy for three test questions on the efficacy of therapeutic sequencing of systemic therapies in PC.
RESULTS AND LIMITATIONS
In this initial evaluation, INSIDE PC outperformed PubMed for question 1 (novel hormonal therapy [NHT] followed by NHT) for the top five, ten, and 20 publications (nDCG score, +43, +33, and +30 percentage points [pps], respectively). For question 2 (NHT followed by poly [adenosine diphosphate ribose] polymerase inhibitors [PARPi]), INSIDE PC and PubMed performed similarly. For question 3 (NHT or PARPi followed by Lu-prostate-specific membrane antigen-617), INSIDE PC outperformed PubMed for the top five, ten, and 20 publications (+16, +4, and +5 pps, respectively).
CONCLUSIONS
We applied INSIDE PC to develop standards for evaluating the performance of AI-based tools for literature extraction. INSIDE PC performed competitively with PubMed and can assist clinicians with therapeutic sequencing in PC.
PATIENT SUMMARY
The medical literature is often very difficult for doctors and patients to search. In this report, we describe INSIDE PC-an artificial intelligence (AI) system created to help search articles published in medical journals and determine the best order of treatments for advanced prostate cancer in a much better time frame. We found that INSIDE PC works as well as another search tool, PubMed, a widely used resource for searching and retrieving articles published in medical journals. Our work with INSIDE PC shows new ways in which AI can be used to search published articles in medical journals and how these systems might be evaluated to support shared decision-making.
PubMed: 38876943
DOI: 10.1016/j.euf.2024.05.022 -
Carbohydrate Polymers Oct 2024The biological activities of heparan sulfate (HS) are intimately related to their molecular weights, degree and pattern of sulfation and homogeneity. The existing...
The biological activities of heparan sulfate (HS) are intimately related to their molecular weights, degree and pattern of sulfation and homogeneity. The existing methods for synthesizing homogeneous sugar chains of low dispersity involve multiple steps and require stepwise isolation and purification processes. Here, we designed a mesoporous metal-organic capsule for the encapsulation of glycosyltransferase and obtained a microreactor capable of enzymatically catalyzing polymerization reactions to prepare homogeneous heparosan of low dispersity, the precursor of HS and heparin. Since the sugar chain extension occurs in the pores of the microreactor, low molecular weight heparosan can be synthesized through space-restricted catalysis. Moreover, the glycosylation co-product, uridine diphosphate (UDP), can be chelated with the exposed metal sites of the metal-organic capsule, which inhibits trans-cleavage to reduce the molecular weight dispersity. This microreactor offers the advantages of efficiency, reusability, and obviates the need for stepwise isolation and purification processes. Using the synthesized heparosan, we further successfully prepared homogeneous 6-O-sulfated HS of low dispersity with a molecular weight of approximately 6 kDa and a polydispersity index (PDI) of 1.032. Notably, the HS generated exhibited minimal anticoagulant activity, and its binding affinity to fibroblast growth factor 1 was comparable to that of low molecular weight heparins.
Topics: Heparitin Sulfate; Polymerization; Anticoagulants; Molecular Weight; Porosity; Humans; Disaccharides; Glycosyltransferases
PubMed: 38876729
DOI: 10.1016/j.carbpol.2024.122297 -
Journal of Dairy Science Jun 2024The aim of the study was to investigate the impact of potassium-based emulsifying salts (ES; 2% wt/wt concentration) with different phosphate chain lengths [dipotassium...
The aim of the study was to investigate the impact of potassium-based emulsifying salts (ES; 2% wt/wt concentration) with different phosphate chain lengths [dipotassium hydrogenphosphate (KHPO; DKP), tetrapotassium diphosphate (KPO; KTPP), pentapotassium triphosphate (KPO; TKPP)] on the physicochemical, viscoelastic, textural, tribological, thermal, and sensory properties of processed cheese (PC; 40% wt/wt dry matter, 50% wt/wt fat in dry matter) during a 60d storage period (6 ± 2°C). On the whole, the hardness of all PC samples increased with the increasing chain length of ES (DKP < TKPP < KTPP) and the prolonging storage period. Moreover, the hardness results were in accordance with those of the rheological analysis. All PC samples exhibited a more elastic character (G' > G"; tan δ < 1). The type of potassium-based ES affected the binding of water into the structure of the PC. Furthermore, the study confirmed that the manufactured PCs received optimal sensory scores, without any excessive bitterness. It could be concluded that the type of applied ES and storage length affected the functional properties of PC. Finally, the information provided in this study could serve as a tool for the dairy industry to help appropriately select potassium-based ES for PC manufacture with desired properties.
PubMed: 38876214
DOI: 10.3168/jds.2024-24939 -
Cell Metabolism Jun 2024Mitochondria house many metabolic pathways required for homeostasis and growth. To explore how human cells respond to mitochondrial dysfunction, we performed...
Mitochondria house many metabolic pathways required for homeostasis and growth. To explore how human cells respond to mitochondrial dysfunction, we performed metabolomics in fibroblasts from patients with various mitochondrial disorders and cancer cells with electron transport chain (ETC) blockade. These analyses revealed extensive perturbations in purine metabolism, and stable isotope tracing demonstrated that ETC defects suppress de novo purine synthesis while enhancing purine salvage. In human lung cancer, tumors with markers of low oxidative mitochondrial metabolism exhibit enhanced expression of the salvage enzyme hypoxanthine phosphoribosyl transferase 1 (HPRT1) and high levels of the HPRT1 product inosine monophosphate. Mechanistically, ETC blockade activates the pentose phosphate pathway, providing phosphoribosyl diphosphate to drive purine salvage supplied by uptake of extracellular bases. Blocking HPRT1 sensitizes cancer cells to ETC inhibition. These findings demonstrate how cells remodel purine metabolism upon ETC blockade and uncover a new metabolic vulnerability in tumors with low respiration.
PubMed: 38876105
DOI: 10.1016/j.cmet.2024.05.014 -
Journal of Medicinal Chemistry Jun 2024P2Y receptor (P2YR) is activated by uridine 5'-diphosphate-glucose, which is involved in many human inflammatory diseases. Based on the molecular docking analysis of...
P2Y receptor (P2YR) is activated by uridine 5'-diphosphate-glucose, which is involved in many human inflammatory diseases. Based on the molecular docking analysis of currently reported P2YR antagonists and the crystallographic overlap study between the reported P2YR antagonist compounds and , a series of N-substituted-acetamide derivatives were designed, synthesized, and identified as novel and potent P2YR antagonists. The most potent antagonist, compound (-(1-benzo[]imidazol-6-yl)-2-(4-bromophenoxy)acetamide, IC = 0.6 nM) without zwitterionic character, showed strong binding ability to P2YR, high selectivity, moderate oral bioactivity, and improved pharmacokinetic profiles. and evaluation demonstrated that compound had satisfactory inhibitory activity on the inflammatory response of monosodium urate (MSU)-induced acute gouty arthritis. decreased inflammatory factor release and cell pyroptosis through the NOD-like receptor family pyrin domain-containing 3 (NLRP3)/gasdermin D (GSDMD) signaling pathway. Thus, compound , with potent P2YR antagonistic activity, and efficacy, and favorable bioavailability ( = 75%), could be a promising lead compound for acute gouty arthritis.
Topics: Acetamides; Humans; Animals; Receptors, Purinergic P2; Molecular Docking Simulation; Mice; Male; Arthritis, Gouty; Structure-Activity Relationship; Purinergic P2 Receptor Antagonists; Drug Discovery; Rats; Crystallography, X-Ray; Rats, Sprague-Dawley; Molecular Structure
PubMed: 38874515
DOI: 10.1021/acs.jmedchem.4c00555 -
Angewandte Chemie (International Ed. in... Jun 2024The thiamine diphosphate (ThDP)-binding motif, characterized by the canonical GDG(X)24-27N sequence, is highly conserved among ThDP-dependent enzymes. We investigated a...
The thiamine diphosphate (ThDP)-binding motif, characterized by the canonical GDG(X)24-27N sequence, is highly conserved among ThDP-dependent enzymes. We investigated a ThDP-dependent lyase (JanthE from Janthinobacterium sp. HH01) with an unusual cysteine (C458) replacing the first glycine of this motif. We found that JanthE has a high substrate promiscuity accepting long aliphatic α-keto acids as donors. Sterically hindered aromatic aldehydes or non-activated ketones are acceptor substrates, giving access to a variety of secondary and tertiary alcohols as carboligation products. The crystal structure solved at a resolution of 1.9 Å reveals that C458 is not primarily involved in the cofactor binding as previously thought for the canonical glycine. Instead, it coordinates methionine 406, thus ensuring the integrity of the active site and the enzyme activity. We further determined the long-sought genuine tetrahedral intermediates formed with pyruvate and 2-oxo-butyrate in the pre-decarboxylation states and unravel atomic details for their stabilization in the active site. Collectively, we unravel an unexpected role for the first residue of the ThDP-binding motif and unlock a family of lyases able to perform valuable carboligation reactions.
PubMed: 38874074
DOI: 10.1002/anie.202404045