Half Inhibitory Concentration Research Articles

A paper-based lateral flow immunochromatographic sensor for the detection of tricyclazole in rice

Tricyclazole is commonly used to prevent rice blast to meet the carbohydrate intake needs of half of the global population, and a large number of toxicological reports indicate that monitoring of tricyclazole is necessary. Here, we analyzed the structure of tricyclazole and designed different hapten derivatization strategies to prepare a high-performance monoclonal antibody (half inhibition concentration of 1.61 ng/mL), and then a lateral flow immunochromatographic sensor based on gold nanoparticles for the detection of tricyclazole in rice, with a limit of detection of 6.74 μg/kg and 13.58 μg/kg in polished and brown rice, respectively. The recoveries in rice were in the range of 84.6–107.4%, no complex pretreatment was required for comparison with LC-MS/MS, and the comparative analysis demonstrated that our method had good accuracy and precision. Therefore, the developed lateral flow immunochromatographic analysis was a reliable and rapid means for the on-site analysis of tricyclazole in rice.

Novel compounds with dual inhibition activity against SARS-CoV-2 critical enzymes RdRp and human TMPRSS2

COVID-19 caused major worldwide problems. The spread of variants and limited treatment encouraged the design of novel anti-SARS-CoV-2 compounds. A series of compounds RH1-23 were designed to dually target RNA-dependent RNA polymerase (RdRp) and transmembrane serine protease 2 (TMPRSS2). Compared to remdesivir, in vitro screening indicated the highest selectivity and potent activity of RH11-13 with half maximum inhibitory concentration (IC50) 3.9, 5.7, and 19.72nM, respectively. RH11-12 showed superior inhibition activity against TMPRSS2 and RdRP with IC50 (1.7 and 4.2), and (6.1 and 4.42) nM, respectively. WaterMap analysis and molecular dynamics studies demonstrated the superior enzyme binding activity of RH11 and RH12. On Vero-E6 cells, RH11 and RH12 significantly inhibited the viral replication with 66 % and 63.2 %, and viral adsorption with 44% and 65%, alongside virucidal effect with 51.40% and 90.5%, respectively. Furthermore, the potent activity of RH12 was tested on TMPRSS2-expressing cells (Calu-3) compared to camostat. RH12 exhibited selectivity index (26.05) similar to camostat (28.01) and comparable to its SI on Vero-E6 cells (22.6). RH12 demonstrated also a significant inhibition of the viral adsorption on Calu-3cells with 60% inhibition at 30nM. The designed compounds exhibited good physiochemical properties. These findings indicate a broad-spectrum antiviral efficacy of the designed compounds, particularly RH12, with a promise for further development.

Effects of the antitumor drugs adagrasib and asciminib on apixaban metabolism in vitro and in vivo

Apixaban is an oral anticoagulant that directly inhibits the target Factor Xa (FXa). In this study, we focused on the in vivo and in vitro effects of adagrasib and asciminib on apixaban metabolism, to discover potential drug-drug interactions (DDI) and explore their inhibitory mechanisms. The levels of apixaban and its metabolite, O-desmethyl-apixaban (M2), were determined by ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). In vitro evaluation, the maximum half inhibitory concentration (IC50) of adagrasib in rat liver microsomes (RLM) and human liver microsomes (HLM) against apixaban was 7.99 μM and 117.40 μM, respectively. The IC50 value of asciminib against apixaban in RLM and HLM was 4.28 μM and 18.42 μM, respectively. The results of the the analysis on inhibition mechanisms showed that adagrasib inhibited the metabolism of apixaban through a non-competitive mechanism, while asciminib inhibited the metabolism of apixaban through a mixed mechanism. Moreover, the interaction of apixaban with adagrasib and asciminib in Sprague-Dawley (SD) rats was also investigated. It was found that the pharmacokinetic characteristics of apixaban were significantly changed when combined with these two antitumor drugs, where AUC(0-t), AUC(0-∞), t1/2, Tmax, and Cmax were increased, while CLz/F was significantly decreased. But both drugs did not appear to affect the metabolism of M2 in a significant way. Consistent results from in vitro and in vivo demonstrated that both adagrasib and asciminib inhibited the metabolism of apixaban. It provided reference data for the future clinical individualization of apixaban.

Impact of altitudinal variation on secondary metabolites, antioxidant, anti-inflammatory and anticancer potential of Adiantum venustum D. Don

Phytoconstituents are being emphasized as chemotherapeutic agents, as well as probable anticancer agents. The purpose of the present research was to assess the antioxidant, anti-inflammatory, and anticancer properties of crude ethanolic extracts and bioactive fractions of Adiantum venustum D. Don collected from three sites in Himachal Pradesh, India. The bioactive phytoconstituents were examined qualitatively and quantitatively in crude ethanolic extracts and fractions of all sites, and biological activities were analyzed. The potential presence of terpenoids, saponins, tannins, phenols, and flavonoids was examined, and it came to light that BF-II [n-butanol fraction of Site-II (2000 m altitude)] had a significantly higher phytochemical concentration. Additionally, the crude ethanolic extracts and fractions were examined for antioxidant and anti-inflammatory properties, and BF-II was found to have potential antioxidant and anti-inflammatory activity, as verified by the IC50 values (lowest half inhibitory concentration) in the DPPH (57.86 ± 0.72 μg/mL), reducing power assay (45.20 ± 0.72 μg/mL), FRAP (73.12 ± 0.23 μM Fe equivalent), BSA (56.76 ± 0.25 μg/mL), and EAA (EAA59.84 ± 0.13 μg/mL). Following that, the anticancer potential was investigated further using the MTT assay on the HeLa cell line, MTT assay revealed that BF-II possesses significantly higher anticancer potential. Furthermore, the dominant crude ethanolic extract and fraction were further investigated using gas chromatography-mass spectroscopy, and high-performance thin layer chromatography techniques from the selected plant. Overall, the study suggests that the n-butanol fraction of A. venustum possesses significantly higher therapeutic value but is still not explored to the extent of its potential.

Cost-efficient and ultrasensitive sensor for electrochemical detection and cytotoxicity assessment of tetracyclines

The uncontrolled overuse of tetracyclines (TCs) has contributed to a significant rise in the levels of not only antibiotic residues in the environmental but also drug- and multidrug-resistant bacteria, necessitating a convenient, sensitive, and rapid assay for the quantitative determination and health risk assessment of TCs. This study presents the development of a nanocomposite sensor which integrates hydroxylated multiwalled carbon nanotubes (MWCNTs-OH), tungsten disulfide quantum dots (WS2QDs), and prussian blue (PB). This sensor was capable of measuring the concentrations of tetracycline (TC), oxytetracycline (OTC), doxycycline (DC), and tigecycline (TEC) at levels ranging from 0.10 to 125.00μM, with detection limits of 0.010, 0.015, 0.020, and 0.060μM, respectively (S/N = 3). The excellent electrochemical performance enabled the sensor to measure TC concentrations in real water samples. Importantly, the sensor was also used to evaluate the cytotoxicity of TCs on human hepatocarcinoma (HepG2) cells. The 48h half-inhibitory concentrations (IC50) for TC, OTC, DC, and TEC were 344.91μM, 365.14μM, 349.59μM, and 604.66μM, respectively, indicating that TC was the most cytotoxic. The sensing platform described herein not only facilitates the quantitative determination of TCs but also allows for the assessment of their cytotoxicity in the water environment, which greatly simplifies the comprehensive evaluation process and reduces the cost, thus holding great promise for TCs testing.

Chemical Profiling of Drimys granadensis (Winteraceae) Essential Oil, and Their Antimicrobial, Antioxidant, and Anticholinesterase Properties.

A complete and comprehensive chemical and biological study of Drimys granadensis, a native Ecuadorian aromatic plant, was conducted. By conventional steam distillation from dried leaves, a yellowish, translucent essential oil (EO) with a density of 0.95 and a refractive index of 1.5090 was obtained. The EO was analyzed by gas chromatography coupled to a mass spectrometer (GC/MS) and an FID detector (GC/FID), respectively. Enantiomeric distribution was also carried out by GC/MS using a chiral selective column (diethyl tert-butylsilyl-BETA-cyclodextrin). The microdilution broth method was employed to assess the antibacterial and antifungal activity of the EO against a panel of opportunistic microorganisms. Antioxidant capacity was measured using diphenyl picryl hydrazyl (DPPH) and azino-bis 3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radicals. Finally, the inhibitory potential of the EO against acetylcholinesterase was also valued. Sixty-four chemical compounds, constituting 93.27% of the total composition, were identified, with major components including γ-muurolene (10.63%), spathulenol (10.13%), sabinene (5.52%), and δ-cadinene (4.22%). The characteristic taxonomic marker of the Drimys genus, Drimenol, was detected at very low percentages (<2%). Two pairs of enantiomers ((1S,5R)-(+)-α-pinene/(1S,5S)-(-)-α-pinene; (1S,5R)-(+)-β-pinene/(1S,5S)-(-)-β-pinene) and one pure enantiomer (1R,4S)-(-)-camphene were identified. Regarding antimicrobial potency, the EO exhibited a significant moderate effect on Listeria monocytogenes with a minimal inhibitory concentration (MIC) value of 250 µg/mL, while with the remaining microorganisms, it exerted less potency, ranging from 500 to 2000 µg/mL. The EO displayed moderate effects against the ABTS radical with a half scavenging capacity of 210.48 µg/mL and no effect against the DPPH radical. The most notable effect was noticed for acetylcholinesterase, with a half inhibition concentration (IC50) of 63.88 ± 1.03 µg/mL. These antiradical and anticholinesterase effects hint at potential pharmacological applications in Alzheimer's disease treatment, although the presence of safrole, albeit in low content (ca. 2%), could limit this opportunity. Further in vivo studies are necessary to fully understand their potential applications.

Isolation of a surfactin-producing strain of Bacillus subtilis and evaluation of the probiotic potential and antioxidant activity of surfactin from fermented soybean meal.

Surfactin, usually produced by microbial metabolism, has many advantages including low toxicity, high biodegradability, and stability at extreme pH levels and temperatures, making it suitable for industry. However, its commercial production has not yet been achieved. A strain with a strong surfactin-producing ability was isolated and identified as Bacillus subtilis SOPC5, based on the appearance of colonies, microscopic observation, and 16S rDNA sequencing. The isolate exhibited significant tolerance to acid, bile, gastric, and intestinal juices, and was sufficiently susceptible to antibiotics. Bacillus subtilis SOPC5 showed high levels of auto-aggregation and surface hydrophobicity, and a strong capacity to secrete protease, amylase, and cellulase. The strainalso exhibited antibacterial activity against Staphylococcus aureus 10 306 with a antibacterial circle diameter of 18.0 ± 1.1 mm. The maximal yield of surfactin (1.32 mg mL-1) was obtained by fermenting soybean meal (SBM) using the isolate under the following conditions: SBM 86 g L-1, inoculation 1.5 × 107 CFU mL-1, FeSO4 1.2 mg L-1, MnSO4 2.6 mg L-1, MgSO4 0.5 mg mL-1, L-Glu 4 mg L-1, temperature 33 °C, duration 120 h, and shaking at 210 rpm. The purity of surfactin was 97.42% as measured by high-performance liquid chromatography (HPLC). The half inhibitory concentration (IC50) values for surfactin to scavenge 1,1-diphenyl-2-picrylhydrazyl (DPPH)and 2,2-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) radical cation (ABTS·+) were 1.275 ± 0.11 and 0.73 ± 0.08 mg mL-1, respectively. This study provides a scientific basis for the application of B. subtilis SOPC5 (as a potential probiotic) and the preparation of its metabolic product (surfactin). © 2024 Society of Chemical Industry.

Systems metabolic engineering of Escherichia coli for high-yield production of Para-hydroxybenzoic acid

Para-hydroxybenzoic acid (PHBA) is extensively used as an additive in the food and cosmetics industries, significantly enhancing product shelf life and stability. While microbial fermentation offers an environment-friendly and sustainable method for producing PHBA, the titer and productivity are limited due to product toxicity and complex metabolic flux distributions. Here, we initially redesigned a L-phenylalanine-producing Escherichia coli by employing rational metabolic engineering strategies, resulting in the production of PHBA reached the highest reported level of 14.17 g/L. Subsequently, a novel accelerated evolution system was devised comprising deaminase, the alpha subunit of RNA polymerase, an uracil-DNA glycosylase inhibitor, and the PHBA-responsive promoter PyhcN. This system enabled us to obtain a mutant strain exhibiting a 47% increase in the half-inhibitory concentration (IC50) for PHBA within 15 days. Finally, the evolved strain achieved a production of 21.35 g/L PHBA in a 5-L fermenter, with a yield of 0.19 g/g glucose and a productivity rate of 0.44 g/L/h. This engineered strain emerges as a promising candidate for industrial production of PHBA through an eco-friendly approach.

Epitope-Shared Hapten Enhancing Antibody Polyreactivity for Simultaneous Immunochromatography of Oxyphenisatin Adulterants.

Given the significant threat posed by oxyphenisatin adulterants (OPHs) in weight-loss foods, simultaneous analysis of the OPHs is necessary. Herein, four novel haptens based on the general epitope shared among the OPHs were raised by computer-aided chemical modeling prediction, with the expectation of eliciting antibody responses targeting three of the OPHs. One obtained monoclonal antibody (mAb) showed maximal half-inhibitory concentration (IC50) of 0.40-12.11 ng/mL for OPHs. The key interaction forces responsible for the corecognition of the OPHs were revealed by the intrinsic molecular mechanism. The developed immunochromatography (ICA) indicated a detection capability for screening (CCβ) for OPHs estimated to be 5-600 ng/g in jelly, candy tablets, and oral liquid. Furthermore, the analysis of 15 real samples by our method showed a good correlation with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Our research not only presented a rapid approach for identifying OPHs adulteration but also proposed an effective hapten prediction strategy to enhance antibody polyreactivity.

Inhibition effect of Cu(II) on nitrogen removal in anammox-denitrification couple system

Due to the wide application in industries, copper can be detected in some nitrogen-rich wastewater. In this research, short-term and long-term experiments were conducted to explore the effects of Cu(II) on the anammox-denitrification couple system. It concluded that the half inhibition concentration (IC50) of Cu(II) was 35.54 mg/L. The system in reactor could tolerate low concentrations of Cu(II) (≤5 mg/L), while the total nitrogen removal efficiency decreased from 93 % to 33 % under 10 mg/L of Cu(II). After 45 days exposure to Cu(II) (1–10 mg/L), 14.54 mg/g SS copper accumulated in the sludge, which largely inhibited the microbial activity. More extracellular polymeric substances (EPS) were secreted to defend against copper toxicity. Proteobacteria (19.18 %–44.04 %) was the dominant phylum and showed excellent tolerance and adaptability to Cu(II). The dominant anammox bacteria, Candidatus_Brocadia, was slightly enhanced under low concentrations of Cu(II), but was highly inhibited under 10 mg/L of Cu(II). PICRUSt2 results showed that some metabolic activities were suppressed under the exposure of copper while defensive responses were also induced. Metabolic disorders eventually led to the death of some microbes, resulting in unrecoverable deterioration in microbial activity. Overall, this study explores the effect of Cu(II) on the anammox-denitrification process and provides a possible inhibition mechanism.

Transient receptor potential vanilloid 1 involved in the analgesic effects of total flavonoids extracted from Longxuejie ().

To evaluate the analgesic effects of total flavonoids of Longxuejie (Resina Dracaenae Cochinchinensis) (TFDB) and explore the possible analgesic mechanism associated with transient receptor potential vanilloid 1 (TRPV1). Whole-cell patch clamp technique was used to observe the effects of TFDB on capsaicin-induced TRPV1 currents. Rat experiments in vivo were used to observe the analgesic effects of TFDB. Western blot and immunofluorescence experiments were used to test the change of TRPV1 expression in DRG neurons induced by TFDB. Results showed that TFDB inhibited capsaicin-induced TRPV1 receptor currents in acutely isolated dorsal root ganglion (DRG) neurons of rats and the half inhibitory concentration was (16.7 ± 1.6) mg/L. TFDB (2-20 mg/kg) showed analgesic activity in the phase Ⅱ of formalin test and (0.02-2 mg per paw) reduced capsaicin-induced licking times of rats. TFDB (20 mg/kg) was fully efficacious on complete Freund's adjuvant (CFA)-induced inflammatory thermal hyperalgesia and capsaicin could weaken the analgesic effects. The level of TRPV1 expressions of DRG neurons was also decreased in TFDB-treated CFA-inflammatory pain rats. All these results indicated that the analgesic effect of TFDB may contribute to their modulations on both function and expression of TRPV1 channels in DRG neurons.

Ergosterol peroxide inducing apoptosis of human hepatocellular carcinoma by regulating mitochondrial apoptosis pathway

This study aims to investigate the effect of ergosterol peroxide(EP) on the apoptosis of human hepatocellular carcinoma and its mechanism of action. The cell viability of HepG2 and SK-Hep-1 cells with 0(blank control), 2.5, 5, 10, 20, 40, and 80 μmol·L~(-1) of EP after 24, 48, and 72 h of action was detected by using CCK-8 assay, and the half inhibitory concentrations(IC_(50)) at 24, 48, and 72 h were calculated. Formal experiments were performed to detect the effect of EP on intracellular reactive oxygen species(ROS) using DCFH-DA staining, the effect of EP on intracellular mitochondrial membrane potential using JC-1 staining, the number of apoptotic cells using Annexin V-FITC/PI double-staining after HepG2 cells were co-cultured with 0(blank control), 10, 20, 40 μmol·L~(-1) EP for 48 h. The effects of EP at different concentrations on apoptotic morphology were detected using AO/EB staining. The effects of different concentrations of EP on the protein expression of mitochondrial apoptosis pathway-related proteins B cell lymphoma 2(Bcl-2), cytochrome C(Cyt-C), Bcl-2-related X protein(Bax), caspase-3, cleaved caspase-3, caspase-9, and cleaved caspase-9 were examined by using Western blot. The results showed that different concentrations of EP could inhibit the proliferation of hepatocellular carcinoma with concentration-and time-dependent trends. Compared with the blank control group, the ROS level in the EP-treated group increased significantly(P&lt;0.05). The mitochondrial membrane potential decreased significantly(P&lt;0.05). The total apoptosis rate increased significantly(P&lt;0.05). The expression of Bcl-2 protein was significantly down-regulated, and the expression of Cyt-C, Bax, cleaved caspase-9, and cleaved caspase-3 were significantly up-regulated(P&lt;0.05). In summary, EP may inhibit the proliferation of hepatocellular carcinoma by modulating the mitochondria-mediated apoptosis pathway and induce apoptosis.

An anoikis-related gene signature predicts prognosis, drug sensitivity, and immune microenvironment in cholangiocarcinoma

BackgroundCholangiocarcinoma is a malignant invasive biliary tract carcinoma with a poor prognosis. Anoikis-related genes are prognostic features of a variety of cancers. However, the value of prognostication and therapeutic effect of anoikis-related genes in cholangiocarcinoma have not been reported. The aim of this research was developing an ARGs signature associated with cholangiocarcinoma patients. MethodsWe introduced transcriptome data to discover genes that were differentially expressed in cholangiocarcinoma. Subsequently, WGCNA was utilized to screen critical module genes in reference to anoikis. The univariate Cox, Lasso regression and Kaplan-Meier survival were executed to build a prognostic signature. We further performed gene functional enrichment, immune microenvironment and immunotherapy analysis between two risk subgroups. Finally, the pRRophetic algorithm was applied to compare the half inhibitory concentration value of several drugs. ResultsA grand total of 1844 genes with differential expression related to the cholangiocarcinoma patients were identified. Furthermore, we obtained 2678 key module genes related to anoikis. Then, a prognostic signature was developed using the 6 prognostic genes (FXYD2, PCBD1, C1RL, GMNN, LAMA4 and HACL1). Independent prognostic analysis showed that risk score and alcohol could function as separate prognostic variables. We found cetain distinction in the immune microenvironment between the two risk subgroups. Moreover, immunotherapy evaluation showed that the anoikis-related gene signature could be applied as a therapy predictor. Finally, Chemotherapeutic drug sensitivity results showed that the low-risk group responded better to bosutinib, gefitinib, gemcitabine, and paclitaxel, while the high-risk group responded better to axitinib, cisplatin, and imatinib. ConclusionThe prognostic signature comprised of FXYD2, PCBD1, C1RL, GMNN, LAMA4 and HACL1 based on anoikis-related genes was established, which provided theoretical basis and reference value for the research and treatment of cholangiocarcinoma.

Effects of Histidine Oligomers in Lipid Nanoparticles on siRNA Delivery.

In this study, we developed histidine oligomer (oHis; 10mer)-incorporating LNPs (H10LNPs) as a novel carrier for efficient siRNA delivery. Notably, the unmodified oHis (10mer) was greatly incorporated within LNPs through ionic interaction with siRNAs, which serves as an endosome escape enhancer. H10LNPs with a size of approximately 65nm demonstrated a significantly enhanced extent of endosomal escape, as evidenced by calcein assay and confocal microscopy images of intracellular fluorescence, surpassing conventional LNPs. Furthermore, the half inhibitory concentration (IC50) of the human endogenous globotriaosylceramide synthase (Gb3 synthase) gene in H10LNPs-treated cells exhibited a significant three-fold decrease, compared to that in LNP-treated cells. Notably, H10LNPs maintained comparable biocompatibility and biodistribution both in vitro and in vivo. Considering that the fabricated siRNA H10LNPs exhibited excellent biocompatibility and superior gene silencing activity over conventional LNPs, these particles can be harnessed for the safe delivery of therapeutic siRNAs. Additionally, this study introduces promising, feasible, simple, and alternative formulation processes for integrating unmodified functional cationic peptides into LNPs to enhance the delivery efficiency of a wide range of nucleic acid-based drugs. This article is protected by copyright. All rights reserved.

An exploratory study on the effect of kynurenine metabolites on sEnd-2 endothelioma cells.

Cancer is the second leading cause of mortality worldwide. The development of anticancer therapy plays a crucial role in mitigating tumour progression and metastasis. Epithelioid hemangioendothelioma is a very rare cancer, however, with a high systemic involvement. Kynurenine metabolites which includel-kynurenine, 3-hydroxykynurenine, 3-hydroxyanthranilic acid and quinolinic acid have been shown to inhibit T-cell proliferation resulting in a decrease in cell growth of natural killercells and T cells. Furthermore, metabolites such as l-kynurenine have been shown to inhibit proliferation of melanoma cells in vitro. Considering these metabolite properties, the present study aimed to explore the in vitro effects of l-kynurenine, quinolinic acid and kynurenic acid on endothelioma sEnd-2 cells and on endothelial (EA. hy926 cells) (control cell line). The in vitro effect at 24, 48, and 72 h exposure to a range of 1-4 mM of the respective kynurenine metabolites on the two cell lines in terms of cell morphology, cell cycle progression and induction of apoptosis was assessed. The half inhibitory concentration (IC50), as determined using nonlinear regression, for l-kynurenine, quinolinic acid and kynurenic acid was 9.17, 15.56, and 535.40 mM, respectively. Optical transmitted light differential interference contrast and hematoxylin and eosin staining revealed cells blocked in metaphase, formation of apoptotic bodies and compromised cell density in l-kynurenine-treated cells. A statistically significant increase in the number of cells present in the sub-G1 phase was observed in l-kynurenine-treated sample. To our knowledge, this was the first in vitro study conducted to investigate the mechanism of action of kynurenine metabolites on endothelioma sEnd-2 cells. It can be concluded that l-kynurenine exerts an antiproliferative effect on the endothelioma sEnd-2 cell line by decreasing cell growth and proliferation as well as a metaphase block. These hallmarks suggest cell death via apoptosis. Further research will be conducted on l-kynurenine to assess the effect on cell adhesion in vitro and in vivo as cell-cell adhesion has been shown to increase metastasis to distant organs therefore, the inhibition of adhesion may lead to a decrease in metastasis.

In Vitro Investigations into the Potential Drug Interactions of Pseudoginsenoside DQ Mediated by Cytochrome P450 and Human Drug Transporters.

Pseudoginsenoside DQ (PDQ), an ocotillol-type ginsenoside, is synthesized with protopanaxadiol through oxidative cyclization. PDQ exhibits good anti-arrhythmia activity. However, the inhibitory effect of PDQ on the cytochrome 450 (CYP450) enzymes and major drug transporters is still unclear. Inhibition of CYP450 and drug transporters may affect the efficacy of the drugs being used together with PDQ. These potential drug-drug interactions (DDIs) are essential for the clinical usage of drugs. In this study, we investigated the inhibitory effect of PDQ on seven CYP450 enzymes and seven drug transporters with in vitro models. PDQ has a significant inhibitory effect on CYP2C19 and P-glycoprotein (P-gp) with a half-inhibitory concentration (IC50) of 0.698 and 0.41 μM, respectively. The inhibition of CYP3A4 and breast cancer-resistant protein (BCRP) is less potent, with IC50 equal to 2.02-6.79 and 1.08 μM, respectively.

Analysis of Quantitative Phytochemical Content and Antioxidant Activity of Leaf, Stem, and Bark of Gymnosporia senegalensis (Lam.) Loes.

To the best of our knowledge, there was no prior report providing valuable preliminary data through a demonstration of the quantitative phytochemical and antioxidant activity of Gymnosporia senegalensis. The total contents of phenols, flavonoid, flavanol, tannin, and saponin were evaluated from different fractions extracted from the leaf, stem, and bark of G. senegalensis by using standards such as gallic acid, quercetin, rutin, tannic acid, and saponin quillaja. The antioxidant potential was measured by 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydrogen peroxide scavenging (H2O2), superoxide anion radical scavenging, metal chelating ferrous ion, ferric reducing antioxidant power (FRAP), and total antioxidant capacity (TAC). Data were subjected to half-inhibitory concentration (IC50) and one-way analysis of variance (ANOVA) at p < 0.05 as a significant value. The total phenol content was found to be highest in the chloroform extract of stem at 97.7 ± 0.02 mg GAE/g. The total flavonoid and flavonol contents in the aqueous extract were 97.1 ± 0.03 mg QE/g and 96.7 ± 0.07 mg RE/g, respectively. The total tannin content in the ethyl acetate extract of leaf was 97.5 ± 0.01 mg TAE/g, and the total saponin content in the methanol extract of stem was 79.1 ± 0.06 mg SQE/g. The antioxidant analysis indicated that IC50 and percentage (%) inhibition were dose-dependent and showed the highest antioxidant activity (40.9 ± 0.9 µg/mL) in methanol extract of leaf for DPPH, (88.8 ± 1.12 µg/mL) in the chloroform extract of stem for H2O2, (43.9 ± 0.15 µg/mL) in the aqueous extract of bark for superoxide anion radical scavenging activity, (26.9 ± 0.11 µg/mL) in the chloroform extract of leaf for the metal chelating ferrous ion activity, (7.55 ± 0.10 mg/mL) in the benzene extract of leaf for FRAP, and (2.97 ± 0.01 mg/mL) in the methanol extract of bark for TAC. These results show that G. senegalensis has great potential in antioxidant activities. The isolation and characterization of specific bioactive compounds and the in vivo applicability of such activity await further extensive studies for drug discovery and development.

Crystal structure of Staphylococcus aureus lipase complex with unsaturated petroselinic acid.

Staphylococcus aureus produces large amounts of toxins and virulence factors. In patients with underlying diseases or compromised immune systems, this bacterium can lead to severe infections and potentially death. In this study, the crystal structure of the complex of S. aureus lipase (SAL), which is involved in the growth of this bacterium, with petroselinic acid (PSA), an inhibitor of unsaturated fatty acids, was determined by X-ray crystallography. Recently, PSA was shown to inhibit S. aureus biofilm formation and the enzymatic activity of SAL. To further characterize the inhibitory mechanism, we determined the half-inhibitory concentration of SAL by PSA and the crystal structure of the complex. The IC50 of the inhibitory effect of PSA on SAL was 3.4 μm. SAL and PSA inhibitors were co-crystallized, and diffraction data sets were collected to 2.19 Å resolution at SPring-8 to determine the crystal structure and elucidate the detailed structural interactions. The results show that the fatty acid moiety of PSA is tightly bound to a hydrophobic pocket extending in two directions around the catalytic residue Ser116. Ser116 was also covalently bonded to the carbon of the unsaturated fatty acid moiety, and an oxyanion hole in SAL stabilized the electrons of the double bond. The difference in inhibitory activity between PSA and ester compounds revealed a structure-activity relationship between SAL and PSA. Additional research is required to further characterize the clinical potential of PSA.

Hapten design, monoclonal antibody preparation and immunoassay development for direct detection of the toxic metabolite of nitrovin without derivatization from fish

Nitrovin, classified as a nitrofuran drug, has been banned in numerous countries due to its carcinogenic effect on human health. However, existing analytical techniques for its detection are laborious, requiring an overnight derivatization process lasting 16 h. In this study, novel haptens were designed to prepare antibodies against aminogunidine (AGD), the metabolite of nitrovin, and then developed an indirect competitive enzyme-linked immunosorbent assay (icELISA) method to detect AGD from fish samples without derivatization for the first time. Computer-assisted molecular modeling predicted the efficiency of novel haptens by conformational studies and electronic analysis. The results were consistent with the predicted results in producing high-affinity monoclonal antibody (mAb) against AGD. The half-inhibition concentration (IC50) value of icELISA was 0.80 ng mL−1 for AGD, with a limit of detection of 0.27 μg kg−1 and recoveries of 89.8%–97.2%. The results showed that this new hapten design and synthesis provided a great prospect to obtain high-affinity mAb for AGD. This developed icELISA substantially avoided the conventional and lengthy derivatization process and was an effective method for the direct detection of AGD in fish tissue without derivatization.

Metallo-Glycodendrimeric Materials against Enterotoxigenic Escherichia coli.

Conjugation of carbohydrates to nanomaterials has been extensively studied and recognized as an alternative in the biomedical field. Dendrimers synthesized with mannose at the end group and with entrapped zero-valent copper/silver could be a potential candidate against bacterial proliferation. This study is aimed at investigating the bactericidal activity of metal-glycodendrimers. The Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) reaction was used to synthesize a new mannosylated dendrimer containing 12 mannopyranoside residues in the periphery. The enterotoxigenic Escherichia coli fimbriae 4 (ETEC:F4) viability, measured at 600 nm, showed the half-inhibitory concentration (IC50) of metal-free glycodendrimers (D), copper-loaded glycodendrimers (D:Cu) and silver-loaded glycodendrimers (D:Ag) closed to 4.5 × 101, 3.5 × 101 and to 1.0 × 10-2 µg/mL, respectively, and minimum inhibitory concentration (MIC) of D, D:Cu and D:Ag of 2.0, 1.5 and 1.0 × 10-4 µg/mL, respectively. The release of bacteria contents onto broth and the inhibition of ETEC:F4 biofilm formation increased with the number of metallo-glycodendrimer materials, with a special interest in silver-containing nanomaterial, which had the highest activity, suggesting that glycodendrimer-based materials interfered with bacteria-bacteria or bacteria-polystyrene interactions, with bacteria metabolism and can disrupt bacteria cell walls. Our findings identify metal-mannose-dendrimers as potent bactericidal agents and emphasize the effect of entrapped zero-valent metal against ETEC:F4.