High-concentrate diets are widely used to enhance growth performance in fattening beef cattle; however, they often compromise rumen epithelial integrity, increasing the risk of rumenitis and systemic inflammation. Supplementation with alkaline mineral complex (AMC) has been shown to alleviate these adverse effects, although the underlying mechanisms remain largely unexplored. In this study, AMC supplementation was associated with improved rumen epithelial integrity and remodeling of the rumen microbiota, characterized by a reduction in Bacteroidota and Prevotella, and an enrichment of Sarcina sp. DSM11001 and Fibrobacter spp., with the latter identified as a key microbial biomarker in the AMC group. Integrated metabolomic and transcriptomic analyses revealed activation of the tryptophan metabolism pathway and accumulation of several anti-inflammatory metabolites, including sulfinpyrazone, Thr-Leu, and 4-guanidinobutyric acid. These metabolomic changes were correlated with the upregulation of tight junction pathways and increased expression of related proteins, which in turn were associated with enhanced epithelial barrier integrity and reduced systemic inflammation in the AMC group. Collectively, these findings suggest that AMC supplementation may protect rumen epithelial integrity by modulating the microbial community and altering ruminal metabolite profiles. This study provides insights into nutritional strategies to prevent epithelial damage under high-concentrate feeding conditions and support the potential use of AMC to maintain rumen health in fattening cattle.

As key members of xenobiotic transcription factors (XFTs), CncC and Maf mediate insecticide sensitivity by regulating detoxification enzyme genes. In this study, with Locusta migratoria as a model, investigations revealed that LmCncC showed particularly high expression in the foregut, midgut, hindgut, and fat bodies. In contrast, LmMaf expression was predominantly localized to the midgut and fat bodies. Functional analysis demonstrated that RNAi-mediated knockdown of LmCncC/LmMaf significantly increased sensitivity to deltamethrin and imidacloprid. To identify the LmCncC- and LmMaf-regulated genes, a comparative transcriptomics analysis was constructed and further identified three key uridine diphosphate-glucuronosyltransferase (UGT) genes downstream of the CncC/Maf pathway. To investigate the role of UGTs in insecticide detoxification, treatment of L. migratoria with sulfinpyrazone, a UGT inhibitor, reduced UGT activity by 42.2% and enhanced sensitivity to imidacloprid and deltamethrin. Among these UGT genes, RNAi-mediated knockdown of LmUGT392C1 significantly enhanced susceptibility to imidacloprid in insecticide bioassays. CRISPR-Cas9-generated LmCncC and LmUGT392C1 mutants validated the essential role of this pathway in imidacloprid detoxification. To clarify the regulatory mechanism of the CncC/Maf signaling pathway on LmUGT392C1, a 2623 bp promoter sequence of LmUGT392C1 was obtained. Dual-luciferase reporter gene assays confirmed that co-overexpression of LmCncC and LmMaf directly activates the promoter of LmUGT392C1. Our study elucidates the CncC/Maf-LmUGT392C1 regulatory axis in insecticide sensitivity, which is applicable to designing new formulations to overcome detoxification or identify potential new resistance mechanisms.

Liver fatty acid binding protein (FABP1) is an abundant cytosolic component that interacts with drugs and can transfer substrates to cytochrome P450 enzymes (P450s, CYPs). FABP1 bound the P450 3A4 substrates diazepam and sulfinpyrazone with Kd values < 2.5 μM. FABP1 weakly attenuated P450 3A4 oxidations of both drugs in two independent assay modes. Reconstitution of human liver microsomes with cytosol attenuated diazepam metabolism but stimulated sulfinpyrazone oxidation, possibly due to GST A1-1. Kinetic modeling of reactions with varying FABP1 and substrate concentrations favored a model of directed substrate transfer to P450 3A4. Kinetic simulations revealed FABP1-dependent inhibition of diazepam oxidation at high physiological concentrations of FABP1, likely due to competition for P450 binding. Consideration of the influence of both P450 and FABP1 on drug metabolism may be critical for accurate modeling and prediction of drug pharmacokinetics.

This study aimed to assessed patients with gout regarding their willingness to pay (WTP) for Human Leukocyte Antigen type-B* 58:01 (HLA-B*58:01) genetic testing before starting allopurinol and identified factors influencing WTP, including the cost of alternative urate-lowering therapy (ULT). A cross-sectional study was conducted using a validated, open-ended questionnaire. Participants over 20 years old were recruited from Naresuan University Hospital and three affiliated sub-district health-promoting hospitals. The majority were male, with a mean age of 64.52 ± 12.82 years. Most participants (86.00%) expressed WTP for the genetic testing, with high cost (62.86%) and insufficient income (25.71%) being the main reasons for un-WTP. Most could afford a median of THB 500 (USD 14.37). WTP for febuxostat, sulfinpyrazone, and benzbromarone was 68.00%, 67.60%, and 71.60%, respectively. Participants were willing to pay a median of THB 1000 (USD 28.74) for febuxostat or sulfinpyrazone, and THB 285 (USD 8.19) for benzbromarone. Multiple logistic regression showed that patient's income and health insurance type particularly non-Civil Servant Medical Benefit Scheme coverage were significantly associated with WTP for the genetic testing. Overall, most participants were willing to pay THB 500 (USD 14.36) for the genetic testing. The study provides insights for policymakers in pricing the genetic testing and ULT coverage in Thailand.

UDP-glucuronosyltransferases (UGTs) catalyze the conjugation of diverse small lipophilic compounds with sugars to form glycosides, playing a critical role in xenobiotic detoxification in insects. In this study, a lambda-cyhalothrin resistant strain (LC-R) of Aphis spiraecola was constructed through continuous selection with lambda-cyhalothrin. Synergism assay revealed that the UGTs inhibitors 5-nitrouracil and sulfinpyrazone dramatically enhanced the toxicity of lambda-cyhalothrin in the resistant A. spiraecola. The activity of UGTs in the LC-R strain was 1.75-fold higher than that in the susceptible strain (SS). Transcriptional analysis further indicated that UGT342A6, UGT342C11, UGT343A33, UGT344B29, UGT344B30, UGT344N14 and UGT350A10 were significantly overexpressed (1.54- to 4.44-fold) in the LC-R strain, among which the transcripts of UGT344B29 and UGT344B30 could be stably induced by lambda-cyhalothrin. Moreover, RNAi-mediated knockdown of UGT344B29 and UGT344B30 via SPc + dsUGT delivery substantially increased sensitivity to lambda-cyhalothrin, reducing the LC50 values to 227.55 mg·L[-1] and 344.47 mg·L[-1], representing decreases of 72.67 % and 58.64 %, respectively, compared with the SPc + dsGFP. Concentration-mortality responses also showed significantly higher mortality in dsUGT-treated aphids across multiple lambda-cyhalothrin concentrations. These results demonstrate that UGT344B29 and UGT344B30 contribute to lambda-cyhalothrin resistance in A. spiraecola. Our findings provide valuable insights for managing pyrethroid resistance of the pest.

The formation of C-C and C-N bonds is an effective means to construct functional complex molecules. Herein, the TsOH-promoted dehydroxylation coupling of diarylmethanols with 1,3-dicarbonyls and sulfonamides to afford a variety of N-alkylated1,3-dicarbonyls and sulfonamides was established with only H2O as the byproduct. Differential 1,3-dicarbonyls and sulfonamides, including pharmaceutical molecules (sulfinpyrazone, phenylbutazone, mofebutazone, celecoxib, topiramate, valdecoxib) were compatible with this system to couple with alcohols in moderate to excellent yields. This method shows the merits of wide substrate scope and mild conditions.

Hyperuricemia, characterized by elevated serum urate levels, is a key factor in the pathogenesis of gout. URAT1 is essential for renal urate reabsorption and has emerged as a critical therapeutic target for managing hyperuricemia. However, the precise transport mechanism and the inhibitory effects of uricosuric drugs on URAT1 remain unclear. Here, we present structures of the double-mutant rat homolog of URAT1 in complex with its substrate urate, and the clinical drugs benzbromarone, lesinurad, verinurad, and sulfinpyrazone. The urate-bound structure elucidates key residues involved in recognizing urate, while the structures bound with drugs clearly demonstrate the distinct binding mode of each drug with URAT1. These drugs stabilize URAT1's inward-facing state, blocking conformational transitions. Additionally, critical interactions essential for its conformational transition are identified. These findings provide a molecular framework for understanding the physiological function of URAT1 and for developing more efficacious therapies to treat hyperuricemia.

Uridine diphosphate glycosyltransferases (UGTs) play critical roles in xenobiotic detoxification and are involved in insecticide resistance. In this study, UGT inhibitors, sulfinpyrazone and 5-nitrouracil, exhibited significant synergistic effects on clothianidin in the clothianidin-resistant strain (CL-R) of Bradysia odoriphaga. UGT enzyme content was significantly higher in the CL-R strain than in the susceptible strain (SS), and both the SS and CL-R strains showed a significant upregulation of UGT content after exposure to clothianidin. Two UGT genes, UGT36M1 and UGT306K1, were significantly overexpressed in the CL-R strain. UGT36M1 was predominantly expressed in the fat body, and UGT306K1 exhibited high levels of expression in the Malpighian tubules and midgut. UGT36M1 and UGT306K1 in the SS strain were significantly upregulated in response to clothianidin exposure. The silencing of UGT36M1 and UGT306K1 significantly enhanced the susceptibility of B. odoriphaga larvae to clothianidin. Furthermore, transgenic overexpression of UGT36M1 and UGT306K1 in Drosophila melanogaster significantly increased the tolerance of fruit flies to clothianidin. These findings provide evidence of the crucial role of UGT36M1 and UGT306K1 in conferring resistance to clothianidin in B. odoriphaga.

SLC17A3 localized to the apical membrane of the renal proximal tubules has been implicated in the urinary excretion of drugs and endogenous/exogenous metabolites transported into the tubules by OAT1 and OAT3. Because SLC17A3 mediates the facilitated diffusion of organic anions, which requires a sensitive and rapid assay, no system has been established to evaluate its transport activity in mammalian cells. In this study, we demonstrated that the exposure of cells expressing click beetle luciferase (bLuc) and SLC17A3 to D-luciferin produces marked bioluminescence, which enables the evaluation of SLC17A3 function. The bioluminescence intensity increased under depolarized membrane potential conditions, consistent with the unique feature of SLC17A3 as a voltage-dependent organic anion transporter. SLC17A3-mediated bioluminescence was saturable with an apparent Michaelis-Menten constant (Km) of 8.1 μM. The inhibitory effects of various compounds including OAT1/OAT3 substrates and inhibitors on bioluminescence were in good agreement with those reported in SLC17A3-expressing Xenopus oocytes using radio-labeled substrates. Interestingly, we found that sulfinpyrazone and lesinurad, uricosuric drugs that inhibit SLC22A12/URAT1, are potent SLC17A3 inhibitors, suggesting the possibility that they alter the pharmacokinetics of OAT1/OAT3-substrate drugs and urate. Taken together, the bioluminescence-based SLC17A3 functional assay is robust and reliable. This strategy enables the study of its transport activity and the identification of potential SLC17A3-mediated drug-drug interactions. This approach also provides an opportunity to elucidate the molecular mechanisms involved in the urinary excretion of organic anions.

Gout is a common form of acute inflammatory arthritis caused by the deposition of monosodium urate crystals within synovium of joints. This leads to severe pain, reducing quality of life for patients with this condition.

The role of insect UDP-glycosyltransferases (UGTs) in the detoxification of insecticides has rarely been reported. A UGT gene UGT2B10 was previously found overexpressed in a fenvalerate-resistant strain of Helicoverpa armigera. Herein, UGT2B10 was cloned, and its involvement in insecticide detoxification was investigated. UGT2B10 was highly expressed in the larvae, mainly in the fat body and midgut. Treatment with UGT inhibitors 5-nitrouracil and sulfinpyrazone significantly enhanced the fenvalerate toxicity. Knocking down UGT2B10 by RNAi significantly increased the larvae mortality by 17.89%. UGT2B10 was further knocked out by CRISPR/Cas9, and a homozygous strain (HD-dUGT2B10) with a C-base deletion at exon 2 was obtained. The sensitivity of HD-dUGT2B10 to fenvalerate, deltamethrin, cyantraniliprole, acetamiprid, and lufenuron increased significantly, with sensitivity index increased 2.523-, 2.544-, 2.250-, 2.473-, and 3.556-fold, respectively. These results suggested that UGT2B10 was involved in the detoxification of H. armigera to insecticides mentioned above, shedding light upon further understanding of the detoxification mechanisms of insecticides by insect UGTs.

Malaria remains one of the highest causes of morbidity and mortality, with 249 million cases and over 608,000 deaths in 2022. Insecticides, which target the Anopheles mosquito vector, are the primary method to control malaria. The widespread nature of resistance to the most important insecticide class, the pyrethroids, threatens the control of this disease. To reverse the stall in malaria control there is urgent need for new vector control tools, which necessitates understanding the molecular basis of pyrethroid resistance. In this study we utilised multi-omics data to identify uridine-diphosphate (UDP)-glycosyltransferases (UGTs) potentially involved in resistance across multiple Anopheles species. Phylogenetic analysis identifies sequence similarities between Anopheline UGTs and those involved in agricultural pesticide resistance to pyrethroids, pyrroles and spinosyns. Expression of five UGTs was characterised in An. gambiae and An. coluzzii to determine constitutive over-expression, induction, and tissue specificity. Furthermore, a UGT inhibitor, sulfinpyrazone, restored susceptibility to pyrethroids and DDT in An. gambiae, An. coluzzii, An. arabiensis and An. funestus, the major African malaria vectors. Taken together, this study provides clear association of UGTs with pyrethroid resistance as well as highlighting the potential use of sulfinpyrazone as a novel synergist for vector control.