Carboxylesterase Research Articles

The diving beetle, Cybister lateralimarginalis (De Geer, 1774), as a bioindicator for subcellular changes affected by heavy metal(loid) pollution in freshwater ecosystems.

The present study assessed Cybister lateralimarginalis as a model species for ecotoxicological studies of metal(loid) bioaccumulation in freshwater ecosystems, by optimising protocols and analysing biomarker activities and metal(loid) concentrations from different sites and investigating their association. To accomplish this, levels of arsenic (As), selenium (Se), cadmium (Cd), mercury (Hg), and lead (Pb) were analysed, as well as a set of biomarkers were evaluated, esterase activity and oxidative stress biomarkers: fluorescence- and glutathione-based. C. lateralimarginalis were sampled in the spring/summer of 2023 along the Sava, Drava and Danube floodplains in Croatia. C. lateralimarginalis samples from Sava floodplain (Mužilovčica) showed altered carboxylesterase (CES) activity, lower glutathione (GSH) levels, and elevated Hg and As concentrations. The Weighted Pollution Index (WPI) indicated moderate pollution at the Sava floodplain, possibly due to agricultural or petrochemical impacts. At the Drava floodplain (Podravlje), higher glutathione S-transferase (GST) activity and elevated Cd, Hg, and As concentrations were observed, likely from soil accumulation, while Stara Drava beetles had lower GSH levels, suggesting agricultural impacts. Despite this, the Drava floodplain showed the lowest WPI. Higher GST activity was noted at the Danube floodplain (Kopački rit and Topoljski Dunavac), with reactive oxygen species (ROS) concentrations (Kopački rit and Podunavlje) possibly reflecting agricultural practices or illegal hunting. The present research successfully applied C. lateralimarginalis as a bioindicator species for assessing metal(loid)s in different environments, highlighting its reliability as an abundant apex predator, emphasizing the significance of integrating biomarker response evaluation into pollutant monitoring for efficient environmental risk assessment in freshwater ecosystems.

Pharmacogenetic Testing for Predicting Methylphenidate Treatment Outcomes in Childhood Attention Deficit Hyperactivity Disorder in Turkey: Focus on Carboxylesterase 1, Latrophilin-3, and Catechol-O-Methyltransferase.

Pharmacogenetic studies involving Carboxylesterase 1 (CES1), Latrophilin-3 (LPHN3), and Catechol-O-methyltransferase (COMT) revealed individual differences regarding therapeutic response in children with attention deficit hyperactivity disorder (ADHD) under methylphenidate (MPH) treatment. This study aimed to evaluate MPH's association with the adverse effect status in children and its relationship with CES1, LPHN3, and COMT in the Turkish population. The study included 102 children and adolescents with ADHD, who were categorized as responders, or the adverse effect group based on their treatment response. The Naranjo Adverse Drug Reaction Probability Scale evaluated the presence and severity of adverse effects. Saliva sample was taken from the patients and genotype distribution of CES1 rs3815583, CES1 rs2307227, LPHN3 rs6551665, LPHN3 rs1947274, LPHN3 rs6858066, LPHN3 rs2345039, and COMT rs4680 were examined. In the adverse effect group, instances of carrying the GG genotype in CES1 rs2307227, having G vs. T genotype and GG vs. GT were significantly higher. In LPHN3 rs2345039, carrying the C genotype vs. G was associated with a serious adverse effect. In COMT rs4680, individuals with the AA or GG genotype were significantly higher in the adverse effect group. Our study suggests a relationship between genetic polymorphisms and the side effect status in children receiving MPH.

CYP2C19 and CES1 gene variants affecting clopidogrel metabolism in a South Asian population from Sri Lanka.

Clopidogrel exhibits substantial variability in therapeutic response, largely contributed by genetic factors. The pharmacogenomic variants data on clopidogrel metabolism in South Asians have been sparsely studied. This study explores the impact of CYP2C19 and CES1 gene variants on clopidogrel metabolism in Sri Lankans, revealing significant pharmacogenomic insights with broader implications for South Asians. Genotype data were filtered out from an anonymized database of 690 Sri Lankans, and minor allele frequencies (MAFs) were calculated. Five variants of CYP2C19 and one variant of CES1 gene were studied. Among the five CYP2C19 variants studied, rs12769205 (A>G) and rs4244285 (G>A) had the highest MAF of 42.1% and 42.0%, respectively. The CES1 variant rs71647871 (C>T) showed a MAF of 0.2%. Sri Lankans exhibited significantly higher MAFs for key variants compared to populations such as Europeans, African Americans, and East Asians (p < 0.05). Given that South Asians share genetic similarities, these findings suggest that a substantial proportion of the region's population may also be poor responders to clopidogrel, increasing the risk of adverse outcomes. This highlights the importance of genotype-guided antiplatelet therapy, which could improve clinical outcomes across South Asia amidst rising cardiovascular disease rates.

Effect of isolated compounds from Combretum trifoliatum on toxicity and detoxification enzymes in Nilaparvata lugens

The brown planthopper (BPH) Nilaparvata lugens (Stål) is a major insect pest of Oryza sativa that causes crop yield loss in tropical regions, including Thailand. In this study, the crude ethanolic extract of the leaves and branches of Combretum trifoliatum , its active isolated components, apigenin and camphor, and Finopril were tested for their ability to control the first to fifth instars of N. lugens. The C. trifoliatum crude extract and both allelochemicals showed insecticide potential (24 h-LC50 ~ 8.83–95.96 mg/L against each instar for crude extract), and their toxicity depended on the time of exposure. Camphor showed the higher efficacy (LD50 ~ 4.43 mg/L) and not different compared to Finopril. All plant compounds tested reduced carboxylesterase (CE) and glutathione-s-transferase (GST) activities. Camphor caused the greatest decreases in CE and GST activities after exposure, whereas apigenin induced a slight change in acetylcholinesterase activity. The results of the present study suggest that C. trifoliatum extract can be used as an insecticide to manage N. lugens populations.

Conjugates of amiridine and salicylic derivatives as promising multifunctional CNS agents for potential treatment of Alzheimer's disease.

New conjugates of amiridine and salicylic derivatives (salicylamide, salicylimine, and salicylamine) with different lengths of alkylene spacers were designed, synthesized, and evaluated as potential multifunctional central nervous system therapeutic agents for Alzheimer's disease (AD). Conjugates demonstrated high acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibition (IC50: AChE, 0.265-4.24 μM; BChE, 0.01-0.64 μM) but poor activity against off-target carboxylesterase (CES). Specifically, conjugates with a (CH2)8 spacer showed the highest AChE and BChE inhibition: 3-16 times more effective than amiridine. Salicylamides 7b and 7c had the maximum BChE/AChE selectivity ratios: 193 and 138, respectively. Conjugates were mixed-type reversible inhibitors of both cholinesterases and displaced propidium from the AChE peripheral anionic site (PAS) at the level of donepezil. All conjugates inhibited Aβ42 self-aggregation in the thioflavin test; inhibition increased with spacer elongation, being greatest for (CH2)8. The results agreed with molecular docking to AChE, BChE, and Aβ42. Conjugates exhibited high 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS)•+-scavenging activity comparable to the standard antioxidant Trolox, and they showed the ability to bind Cu2+, Fe2+, and Zn2+. Conjugates had favorable predicted intestinal absorption and blood-brain barrier permeability. Altogether, the results indicate that the new conjugates possess potential for further development as multifunctional anti-AD drug candidates.

Inter- and intraspecific blood-related biomarkers and chemical exposure in confined and free-living sea turtles.

The relevance of recovery centers and head-starting programs for rescue, rehabilitation, rearing, and conservation of sea turtles is recognized worldwide. In addition, these centers contribute to generating biochemical and physiological data needed to identify health markers and provide baseline values. Because of the marine ecosystems' deterioration, biomarker identification is a global priority for sea turtle conservation; nevertheless, information on specific endpoints, such as neurotoxicity and mutagenesis, is still limited in sea turtles. This study aimed to contrast a set of non-invasive blood biomarkers with ecotoxicological and clinical applications in confined green sea turtles (Chelonia mydas) compared with free-living ones from the Mexican Caribbean. Additionally, interspecific (green, hawksbill (Eretmochelys imbricata), loggerhead (Caretta caretta) turtles) differences were also evaluated. Plasmatic organochlorine pesticides (OCs) and polychlorinated biphenyl (PCBs) were also determined. The concentration ranges of uric acid, total proteins, lipids (cholesterol and triglycerides), and thyroxine of both confined and free-living green turtles fell outside the reference intervals for the species. Additionally, confined green turtles had the highest number of erythrocytic nuclear abnormalities (ENA) and elevated levels of hemoglobin, lipid peroxidation, and activity of glutathione S-transferase, glutathione peroxidase, and carboxylesterase (CE). Contrasts among confined species identified hawksbill turtles with the lowest glutathione reductase activity, green turtles with the lowest ENA frequency and CE activity, and loggerhead turtles with the highest plasmatic concentrations of PCBs and OCs. The information here provided can be used as information in health monitoring programs and for conservation and management policies at regional, national, and international level.

Pomegranate juice treatment reverses carbon tetrachloride (CCl4)-induced increased acetylcholinesterase activity and cell death via suppression of oxidative stress in rats

BackgroundEnvironmental pollution, including exposure to carbon tetrachloride (CCl4), poses serious health risks, particularly through oxidative stress, which may lead to neurodegenerative damage. Antioxidants, especially those found in natural products, show potential in mitigating these toxic effects. Pomegranate juice (PJ), rich in bioactive phytochemicals, has demonstrated antioxidant, anti-inflammatory, and neuroprotective properties.ObjectiveThis study aimed to investigate the protective effects of PJ on neurotoxicity induced by CCl4 in rats, assessing specific markers of oxidative stress, enzymatic activity, and apoptotic cell death.Material and MethodsTwenty-eight male Wistar rats were divided into four groups: Control, CCl4, PJ, and CCl4+PJ. The CCl4 group received intraperitoneal injections of CCl4 (0.2 ml/100 g) twice weekly for six weeks, while the PJ group received PJ orally (4 ml/kg) daily for 30 days. The CCl4+PJ group received both treatments in sequence. Brain tissues were analysed for malondialdehyde (MDA), reduced glutathione (GSH), acetylcholinesterase (AChE), glutathione S-transferase (GST), glutathione reductase (GR), and carboxylesterase (CaE) activity. Apoptotic cell death was assessed using TUNEL staining.ResultsCCl4 exposure resulted in a marked increase in MDA levels and AChE activity in brain tissue (p&lt;0.05), alongside a significant decrease in reduced GSH levels and GST activity (p&lt;0.05). Treatment with PJ significantly lowered MDA levels and AChE activity in the CCl4+PJ group compared to the CCl4 group (p&lt;0.05). However, GSH levels and GST activity showed no significant changes in the CCl4+PJ group. TUNEL staining indicated a reduction in apoptotic cells in the CCl4+PJ group versus the CCl4 group, suggesting reduced cellular damage with PJ treatment (p&lt;0.05).ConclusionsPJ demonstrates neuroprotective potential against CCl4-induced oxidative stress and neurotoxicity in rats by reducing oxidative markers and apoptosis. These findings suggest that PJ could serve as a natural protective agent against neurodegenerative risks associated with environmental pollutants like CCl4.

Key genes in a “Galloylation-Degalloylation cycle” controlling the synthesis of hydrolyzable tannins in strawberry plants

Abstract Strawberry fruits, known for their excellent taste and potential health benefits, are particularly valued for their rich content of hydrolyzable tannins (HTs). These compounds play key roles in regulating growth and development. However, the molecular mechanisms underlying HT synthesis in plants remains poorly elucidated. In this study, based on a correlation analysis between the transcriptome and metabolome of HTs, galloyl glucosyltransferase (UGT84A22), serine carboxypeptidase-like acyltransferases (SCPL-ATs), and carboxylesterases (CXEs) were screened. Furthermore, in vitro enzymatic assays confirmed that FaSCPL3–1 acted as a hydrolyzable tannins synthase (HTS), catalyzing the continuous galloylation of glucose to form simple gallotannins (GTs). Additionally, FaCXE1/FaCXE3/FaCXE7 catalyzed the degalloylation of simple GTs and ellagitannins (ETs), and FaUGT84A22 catalyzed the glycosylation of gallic acid (GA) to produce 1-O-β-glucogallin (βG), a galloyl donor. Moreover, in FvSCPL3–1-RNAi transgenic strawberry plants, the contents of simple GT and some ET compounds were reduced, whereas, in FaCXE7 overexpressing strawberry plants, these compounds were increased. These enzymes constituted a biosynthetic pathway of galloyl derivatives, termed the “galloylation-degalloylation cycle” (G-DG cycle). Notably, the overexpression of FaCXE7 in strawberry plants not only promoted HT synthesis but also interfered with plant growth and development by reducing lignin biosynthesis. These findings offer new insights into the mechanisms of HT accumulation in plants, contributing to improving the quality of berry fruits quality and enhancing plant resistance.

Impaired chaperone-mediated autophagy leads to abnormal SORT1 (sortilin 1) turnover and CES1-dependent triglyceride hydrolysis

ABSTRACT SORT1 (sortilin 1), a member of the the Vps10 (vacuolar protein sorting 10) family, is involved in hepatic lipid metabolism by regulating very low-density lipoprotein (VLDL) secretion and facilitating the lysosomal degradation of CES1 (carboxylesterase 1), crucial for triglyceride (TG) breakdown in the liver. This study explores whether SORT1 is targeted for degradation by chaperone-mediated autophagy (CMA), a selective protein degradation pathway that directs proteins containing KFERQ-like motifs to lysosomes via LAMP2A (lysosomal-associated membrane protein 2A). Silencing LAMP2A or HSPA8/Hsc70 with siRNA increased cytosolic SORT1 protein levels. Leupeptin treatment induced lysosomal accumulation of SORT1, unaffected by siLAMP2A co-treatment, indicating CMA-dependent degradation. Human SORT1 contains five KFERQ-like motifs (658VVTKQ662, 730VREVK734, 733VKDLK737, 734KDLKK738, and 735DLKKK739), crucial for HSPA8 recognition; mutating any single amino acid within these motifs decreased HSPA8 binding. Furthermore, compromised CMA activity resulted in elevated SORT1-mediated degradation of CES1, contributing to increased lipid accumulation in hepatocytes. Consistent with in vitro findings, LAMP2A knockdown in mice exacerbated high-fructose diet-induced fatty liver, marked by increased SORT1 and decreased CES1 levels. Conversely, LAMP2A overexpression promoted SORT1 degradation and CES1D accumulation, counteracting fasting-induced CES1D suppression through CMA activation. Our findings reveal that SORT1 is a substrate of CMA, highlighting its crucial role in directing CES1 to lysosomes. Consequently, disrupting CMA-mediated SORT1 degradation significantly affects CES1-dependent TG hydrolysis, thereby affecting hepatic lipid homeostasis. Abbreviations: APOB: apolipoprotein B; CES1: carboxylesterase 1; CMA: chaperone-mediated autophagy; HSPA8/Hsc70: heat shock protein family A (Hsp70) member 8; LAMP2A: lysosomal associated membrane protein 2A; LDL-C: low-density lipoprotein-cholesterol; PLIN: perilipin; SORT1: sortilin 1; TG: triglyceride; VLDL: very low-density lipoprotein; Vps10: vacuolar protein sorting 10.

Enhancing the photo-induced oxidase-like activity of fluorescein with methyl viologen for colorimetric detection of organophosphorus pesticide

Overcoming the intrinsic low activity of most photoinduced oxidase mimics has been extremely challenging. In this work, we developed a methyl viologen (MV2+) mediated strategy to enhance the oxidase-like activity of fluorescein. The presence of MV2+ gives it a high affinity for TMB with a low Michaelis-Menten constant (Km) of 0.053 mM, which is about 2.8 times lower than that of fluorescein and with a remarkable catalytic constant (Kcat) as 0.2490 s−1, which is 3 times as high as that of fluorescein. Fluorescein diacetate (FDA) without oxidase-like activity can be hydrolyzed in situ to produce fluorescein in the presence of carboxylesterases (CaE). Based on the inhibition of CaE activity by organophosphorus pesticides (OP), a novel colorimetric signal biosensor was established with a wide linear range from 1.0 to 200 ng/mL. This work not only provides a convenient and feasible strategy for enhancing the activity of photoinduced oxidase mimics but also blazes a new pathway for the sensitive detection of OP.

Bivariate tracking of NIR phototherapeutic probe that illuminates the deterioration process of NAFLD-HCC

Non-alcoholic fatty liver disease (NAFLD) has evolved to become a major cause of hepatocellular carcinoma (HCC). Visualization of NAFLD-HCC deterioration process imaging is essential to understand the underlying pathophysiological processes. However, currently relevant probes with short emission wavelengths, univariate and the inability to achieve theranostics functionality have encountered obstacles in further evaluating the NAFLD-HCC process. Here, we present a carboxylesterase (CE)-activated NIR fluorescent probe (BODJ) which has lipid droplets (LDs)-targeting ability and emits at a wavelength of 858 nm with a fluorescence quantum yield of 19.06%. CE-activated BODJ was used as a visual tool to successfully visualize both NAFLD deterioration processes and HCC in situ based on changes in the average number of LDs and the associated fluorescence intensity fluctuations. Imaging results showed that changes associated with CE and LDs in the modelled cells varied during the transition from nonalcoholic fatty liver to nonalcoholic steatohepatitis and later progression to HCC, highlighting the close association between bivariate and disease. We also demonstrate that BODJ has photodynamic (PDT) and photothermal therapy (PTT) capabilities, allowing image-guided dual phototherapy to damage HCC in situ. This NIR probe, which takes advantage of bivariate to track the deterioration process that illuminates NAFLD-HCC and has dual phototherapy capabilities, provides new ideas for the design of probes related to the diagnosis and treatment of hepatic metabolic diseases.

Stress indicators in conservative tissues of Humboldt penguin under captivity

Humboldt penguins (Spheniscus humboldti) from the Barcelona zoo (n=9) were followed to assess their physiological stress status using conservative protocols. Corticosterone levels were measured in feathers and plasma as indicative of chronic and acute physiological stress, respectively. Other markers: B-esterases, potentially indicative of xenobiotic exposure were measured in plasma of these same individuals and reported for the first time in this species. The sensitivity to chemicals of environmental concern, employed as plastic additives, was assessed in vitro with plasma of this species using the inhibition of carboxylesterase (CE) and acetylcholinesterase enzymatic measurements. Among the tested additives, the organophosphorus flame retardants displayed the highest in vitro inhibitory potential on basal CE activity, suggesting their potential utility as biomarkers of this particular chemical class. Additionally, enzymatic measurements in plasma are determined for the first time in Humboldt penguin and can be regarded as baseline values for a potential field monitoring application.

Carboxylesterase-activatable multi-in-one nanoplatform for near-infrared fluorescence imaging guided chemo/photodynamic/sonodynamic therapy toward cervical cancer

Traditional tumor treatment faces great challenge owning to inherent drawbacks. Activatable prodrugs with multi-modality therapeutic capacity are highly desired. In this consideration, a responsiveness-released multi-in-one nanoplatform, PLGA-PEG@HC, toward cervical cancer therapy was innovatively developed. Among the nanoplatform, HC was constructed by incorporating chlorambucil, a classic chemotherapy drug into a near-infrared photo- and sono-sensitizer, HCH via ester linker, which can be specifically hydrolyzed by carboxylesterase (CES). HC is scarcely fluorescent and toxic due to the caging of HCH and chlorambucil, thus achieving low background signal and minimal side effects. However, once selectively hydrolyzed by tumor enriched CES, ester bond will be broken. Consequently, HCH and chlorambucil are released so as to achieve near-infrared fluorescence imaging and synergistic photodynamic/sonodynamic/chemo therapy. PLGA-PEG packaging ensures the biocompatibility of HC. The as-obtained nanoplatform, with diameter of 97 nm, achieves tumor targeting capacity via EPR. In vitro and in vivo applications have demonstrated that PLGA-PEG@HC can accumulate in tumor tissues, exhibit CES-activatable near-infrared fluorescence imaging and efficient tumor suppression capacity. Compared with the reported combinational therapy materials which are complex in compositions, PLGA-PEG@HC is simple in formulation but demonstrates near-infrared fluorescence traced and considerable therapy efficacy toward tumors, which may accelerate the clinical translation.

Population pharmacokinetic study of the effect of polymorphisms in the ABCB1 and CES1 genes on the pharmacokinetics of dabigatran.

The impact of genetic polymorphisms in the ABCB1 and CES1 genes on dabigatran plasma concentrations remains a subject of debate, and the purpose of this study was to quantitatively assess the effects of genetic polymorphisms on dabigatran esters in healthy Chinese subjects employed a population pharmacokinetic (PopPK) approach. In total, 1,926 pharmacokinetic (PK) samples from 123 healthy individuals who were given 150mg of dabigatran orally during a fasting state or postprandially were analyzed using the PopPK model. A two-compartment model with first-order absorption was found to adequately describe the PK data. The results showed that covariates food intake and ABCB1 SNP rs4148738 were shown to have statistically significant impacts. Specifically, in postprandial administration increased lag time (ALAG) and clearance (CL) by 2.65% and 0.51%, respectively, and decreased absorption rate constant (KA) by 0.24%. Additionally, in subjects with CT genotype ABCB1 (rs4148738), the central ventricular volume of distribution (V2) was increased by 0.38%. In summary, the PopPK model developed in this study was robust and effectively characterized the pharmacokinetics of dabigatran in healthy Chinese adults, demonstrating that both food and ABCB1 genetic variation significantly influence the absorption and plasma concentration levels of dabigatran.

Intestinal human carboxylesterase 2 (CES2) expression rescues drug metabolism and most metabolic syndrome phenotypes in global Ces2 cluster knockout mice.

Carboxylesterase 2 (CES2) is expressed mainly in liver and intestine, but most abundantly in intestine. It hydrolyzes carboxylester, thioester, and amide bonds in many exogenous and endogenous compounds, including lipids. CES2 therefore not only plays an important role in the metabolism of many (pro-)drugs, toxins and pesticides, directly influencing pharmacology and toxicology in humans, but it is also involved in energy homeostasis, affecting lipid and glucose metabolism. In this study we investigated the pharmacological and physiological functions of CES2. We constructed Ces2 cluster knockout mice lacking all eight Ces2 genes (Ces2-/- strain) as well as humanized hepatic or intestinal CES2 transgenic strains in this Ces2-/- background. We showed that oral availability and tissue disposition of capecitabine were drastically increased in Ces2-/- mice, and tissue-specifically decreased by intestinal and hepatic human CES2 (hCES2) activity. The metabolism of the chemotherapeutic agent vinorelbine was strongly reduced in Ces2-/- mice, but only marginally rescued by hCES2 expression. On the other hand, Ces2-/- mice exhibited fatty liver, adipositis, hypercholesterolemia and diminished glucose tolerance and insulin sensitivity, but without body mass changes. Paradoxically, hepatic hCES2 expression rescued these metabolic phenotypes but increased liver size, adipose tissue mass and overall body weight, suggesting a "healthy" obesity phenotype. In contrast, intestinal hCES2 expression efficiently rescued all phenotypes, and even improved some parameters, including body weight, relative to the wild-type baseline values. Our results suggest that the induction of intestinal hCES2 may combat most, if not all, of the adverse effects of metabolic syndrome. These CES2 mouse models will provide powerful preclinical tools to enhance drug development, increase physiological insights, and explore potential solutions for metabolic syndrome-associated disorders.

Impact of Genetic Polymorphisms and Drug-Drug Interactions Mediated by Carboxylesterase1 on Remimazolam Deactivation.

Remimazolam (Byfavo®), a recent FDA-approved ester-linked benzodiazepine, offers advantages in sedation, such as rapid onset and predictable duration, making it suitable for broad anesthesia applications. Its favorable pharmacological profile is primarily attributed to rapid hydrolysis, the primary metabolism pathway for its deactivation. Thus, understanding remimazolam hydrolysis determinants is essential for optimizing its clinical use. This study aimed to identify the enzyme(s) and tissue(s) responsible for remimazolam hydrolysis and to evaluate the influence of genetic polymorphisms and drug-drug interactions (DDIs) on its hydrolysis in the human liver. An initial incubation study with remimazolam and phosphate buffer saline (PBS), human serum, and the S9 fractions of human liver and intestine demonstrated that remimazolam was exclusively hydrolyzed by human liver S9 fractions. Subsequent incubation studies utilizing a Carboxylesterase inhibitor (Bis-para-nitrophenylphosphate, BNPP), recombinant human Carboxylesterase1 (CES1) and Carboxylesterase 2 (CES2) confirmed that remimazolam is specifically hydrolyzed by CES1 in human liver. Furthermore, in vitro studies with wild-type CES1 and CES1 variants transfected cells revealed that certain genetic polymorphisms significantly impair remimazolam deactivation. Notably, the impact of CES1 G143E was verified using individual human liver samples. Moreover, our evaluation of the DDIs between remimazolam and several other substrates/inhibitors of CES1-including simvastatin, enalapril, clopidogrel and sacubitril- found that clopidogrel significantly inhibited remimazolam hydrolysis at clinically relevant concentrations, with CES1 genetic variants potentially influencing the interactions. In summary, CES1 genetic variants and its interacting drugs are crucial factors contributing to interindividual variability in remimazolam hepatic hydrolysis, holding the potential to serve as biomarkers for optimizing remimazolam use. Significance Statement This investigation demonstrates that remimazolam is deactivated by CES1 in the human liver, with CES1 genetic variants and DDIs significantly influencing its metabolism. These findings emphasize the need to consider CES1 genetic variability and potential DDIs in remimazolam use, especially in personalized pharmacotherapy to achieve optimal anesthetic outcomes.

Rational Design of an Activatable Near-Infrared Fluorogenic Platform for In Vivo Orthotopic Tumor Imaging and Resection.

Rational and effective design of a universal near-infrared (NIR) light-absorbed platform employed to prepare diverse activatable NIR fluorogenic probes for in vivo imaging and the imaging-guided tumor resection remains less exploited but highly meaningful. Herein, mandelic acid with a core structure of 4-hydroxylbenzyl alcohol to link recognition unit, a fluorophore and a quencher was employed to prepare activatable probes. We exemplified ester as carboxylesterase (CE)-recognized unit, ferrocene as quencher and phenothiazinium as NIR fluorophore to afford fluorogenic probes termed NBS-Fe-CE and NBS-C-Fe-CE. These probes enabled the conversion toward CE with significant fluorescence increases and successfully discriminate CE activity in cells. NIR light enhances the tumor penetration and enable imaging-guided orthotopic tumor resection. This specific case demonstrated that this platform can be effectively used to construct diverse NIR probes for imaging analytes in biological systems.

Rational Design of an Activatable Near‐Infrared Fluorogenic Platform for In Vivo Orthotopic Tumor Imaging and Resection

Rational and effective design of a universal near‐infrared (NIR) light‐absorbed platform employed to prepare diverse activatable NIR fluorogenic probes for in vivo imaging and the imaging‐guided tumor resection remains less exploited but highly meaningful. Herein, mandelic acid with a core structure of 4‐hydroxylbenzyl alcohol to link recognition unit, a fluorophore and a quencher was employed to prepare activatable probes. We exemplified ester as carboxylesterase (CE)‐recognized unit, ferrocene as quencher and phenothiazinium as NIR fluorophore to afford fluorogenic probes termed NBS‐Fe‐CE and NBS‐C‐Fe‐CE. These probes enabled the conversion toward CE with significant fluorescence increases and successfully discriminate CE activity in cells. NIR light enhances the tumor penetration and enable imaging‐guided orthotopic tumor resection. This specific case demonstrated that this platform can be effectively used to construct diverse NIR probes for imaging analytes in biological systems.

Phytochemical screening and toxicity assessment of compounds isolated from the leaves of Mangifera indica L. for the control of Spodoptera litura (Lepidoptera; Noctuidae)

This study aimed to analyze the phytochemical composition and antioxidative capabilities of mango leaves (Mangifera indica L., Nam Dok Mai), indigenous to Sa Kaeo Province, Thailand. Various solvents with differing polarities, including n-hexane, DCM, ethyl acetate, and MeOH, were utilized for leaf extraction. The findings revealed the existence of eight groups of phytochemical compounds: alkaloids, flavonoids, coumarins, saponins, tannins, terpenoids, steroids, and cardiac glycosides. The MeOH crude extract exhibited the highest concentration of total phenolic compounds at 409.88 ±0.02 mg GAE/g. Furthermore, the MeOH crude extract demonstrated the strongest antioxidant activity, with an IC50 value of 0.52 ±0.02 μg/ml, as determined by the DPPH method. High-performance liquid chromatography (HPLC) was employed to identify gallic acid and mangiferin in the MeOH crude extract. Laboratory tests were conducted using the topical application method to evaluate the toxicity of the M. indica leaf crude extract on 2nd instar Spodoptera litura larvae. The MeOH crude extract exhibited high efficacy, with an LD50 value of 10.58 µg per larvae within 24 hours. Gallic acid and mangiferin were identified as the primary active ingredients, with LCD50 values of 1.19 µg per larvae and 1.90 µg per larvae, respectively, within 24 hours. Additionally, the impact on detoxification enzymes (24 hours post-treatment) was assessed in surviving 2nd instar S. litura larvae using the topical application method. The MeOH extract treatment resulted in 1.31-fold inhibition of carboxylesterase (CE), 1.31-fold inhibition of glutathione-S-transferase (GST), and 1.32-fold inhibition of acetylcholinesterase (AChE).

Detection of carboxylesterases by an activatable NIR fluorescence probe with high selectivity in living systems

Carboxylesterases (CEs) have attracted increasingly attention in the physiological and pathological processes of many diseases such as diabetes, hepatocellular carcinoma (HCC) and drug metabolism. Selective detection of CEs activity is imperative to evaluate the pathophysiological process of CEs-related disease. Herein, a selective near-infrared (NIR) fluorescent probe (DCM-CE) with high selectivity is reported. DCM-CE was composed of dicyanoisophorone-based fluorophore and carbamate unit which is a highly selective identification group for CEs. DCM-CE exhibited good sensitivity for CEs detection at physiological pH and temperature. Furthermore, DCM-CE featured large Stokes shift (175 nm) and good at tracking the drug-induced trace change of CEs activity in HepG2 cells with little effect on cell viability. Moreover, DCM-CE was applied to monitor the activity of CEs in living mice. Taken all together, DCM-CE had extensive potential application value in detecting CEs for evaluating related diseases.