Carboxylesterase Enzymes Research Articles

Harnessing Pillar[5]arene Host-Guest Complexation To Improve pH Stability and Affect Enzymatic Degradation of the Anticancer Prodrug Capecitabine: A 19 FNMR Study.

Cancer is a global health problem, and supramolecular chemotherapy is emerging as a novel strategy to battle the disease. Here, we first evaluated the thermodynamic and kinetic stability of the complexes formed between several water-soluble per-substituted pillar[5]arene derivatives and capecitabine (1), a widely used oral chemotherapeutic prodrug. The exchange rate was studied, for the first time in pillararene chemistry, by the 19 F guest exchange saturation transfer (GEST) NMR technique. Importantly, when we evaluated the effect of complexation on the characteristics of 1, we found that the complexation of 1 with such pillar[5]arene hosts increased capecitabine stability at acidic pH very significantly and slowed its enzymatic degradation by the carboxylesterase enzyme in a manner that depended on the host. These interesting findings could have implications on the clinical use of this heavily used prodrug and might affect the management of cancer patients.

Cell-Dependent Activation of ProTide Prodrugs and Its Implications in Antiviral Studies.

The ProTide prodrug design is a powerful tool to improve cell permeability and enhance the intracellular activation of nucleotide antiviral analogues. Previous in vitro studies showed that the activation of ProTide prodrugs varied in different cell lines. In the present study, we investigated the activation profiles of two antiviral prodrugs tenofovir alafenamide (TAF) and sofosbuvir (SOF) in five cell lines commonly used in antiviral research, namely, Vero E6, Huh-7, Calu-3, A549, and Caco-2. We found that TAF and SOF were activated in a cell-dependent manner with Vero E6 being the least efficient and Huh-7 being the most efficient cell line for activating the prodrugs. We also demonstrated that TAF was activated at a significantly higher rate than SOF. We further analyzed the protein expressions of the activating enzymes carboxylesterase 1, cathepsin A, histidine triad nucleotide-binding protein 1, and the relevant drug transporters P-glycoprotein and organic anion-transporting polypeptides 1B1 and 1B3 in the cell lines using the proteomics data extracted from the literature and proteome database. The results revealed significant differences in the expression patterns of the enzymes and transporters among the cell lines, which might partially contribute to the observed cell-dependent activation of TAF and SOF. These findings highlight the variability of the abundance of activating enzymes and transporters between cell lines and emphasize the importance of selecting appropriate cell lines for assessing the antiviral efficacy of nucleoside/nucleotide prodrugs.

Ferulic acid production from wheat bran by integration of enzymatic pretreatment and a cold-adapted carboxylesterase catalysis

High-value chemical production from natural lignocellulose transformation is a reliable waste utilization approach. A gene encoding cold-adapted carboxylesterase in Arthrobacter soli Em07 was identified. The gene was cloned and expressed in Escherichia coli to obtain a carboxylesterase enzyme with a molecular weight of 37.2 KDa. The activity of the enzyme was determined using α-naphthyl acetate as substrate. Results showed that the optimum enzyme activity of carboxylesterase was at 10 °C and pH 7.0. It was also found that the enzyme could degrade 20 mg enzymatic pretreated de-starched wheat bran (DSWB) to produce 235.8 μg of ferulic acid under the same conditions, which was 5.6 times more than the control. Compared to the chemical strategy, enzymatic pretreatment is advantageous because it is environmentally friendly, and the by-products can be easily treated. Therefore, this strategy provides an effective method for high-value utilization of biomass waste in agriculture and industry.

Failure of methanol detoxification in pests confers broad spectrum insect resistance in PME overexpressing transgenic cotton

Methanol is noxious to insect pests, but most plants do not make enough of it to shield themselves from encroaching insects. Methanol emission is known to increase in the instance of herbivory. In the current study, we showed that Aspergillus niger pectin methylesterase over-expression increases methanol emission and confers resistance to polyphagous insect pests on transgenic cotton plants by impeding the possible methanol detoxification pathways. Transgenic plants emitted ∼11 fold higher methanol displaying insect mortality of 96% and 93% in Helicoverpa armigera and Spodoptera litura, respectively. The larvae were unable to survive and finish their life cycle and the surviving larvae exhibited severe growth retardation. Insects try to detoxify methanol via catalase, carboxylesterase and cytochrome P450 monooxygenase enzymes, amongst which cytochrome P450 plays a major role in oxidizing methanol to formaldehyde and formaldehyde to formic acid, which is broken down into carbon dioxide and water. In our study, catalase and esterase enzymes were found to be upregulated, but cytochrome P450 monooxygenase levels were not much affected. Leaf disc assays and In-planta bioassays also showed 50–60% population reduction in the sap sucking pests, such as Bemisia tabaci and Phenacoccus solenopsis. These findings imply that elevated methanol emissions confer resistance in plants against chewing and sap-sucking pests by tampering the methanol detoxification pathways. Such mechanism will be useful in imparting expansive resistance against pests in plants.

Cypermethrin toxicity in the environment: analytical Insight into detection methods and microbial degradation pathways.

The unrestricted utilization of xenobiotic compounds has sparked widespread concern by the world's growing population. A synthetic pyrethroid called cypermethrin (CP) is commonly utilized as an insecticide in horticulture, agriculture, and pest control. The high toxicity levels of accumulated CP have prompted environmental concerns; it damages soil fertility, and an ecosystem of essential bacteria, and causes allergic reactions and tremors in humans by affecting their nervous systems. The damage caused by CP to groundwater, food, and health makes it imperative that new effective and sustainable alternatives are investigated. Microbial degradation has been established as a reliable technique for mineralizing CP into less toxic chemicals. Among the many enzymes produced by bacteria, carboxylesterase enzymes are determined to be the most efficient in the CP breakdown process. High-performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS) has been reported as the best methods for determining CP and its metabolized products, with detection limits as low as ppb from diverse environmental samples. The current study describes the ecotoxicological impact of CP and innovative analytical techniques for their detection. The newly isolated CP-degrading bacterial strains have been evaluated in order to develop an efficient bioremediation strategy. The proposed pathways and the associated critical enzymes in the bacterial mineralization of CP have also been highlighted. Additionally, the strategic action to control CP toxicity has been discussed.

Study on Characterization and Pesticidal Evaluation of Diterpenes Isolated from Chloroform Extract of Hedychium spicatum Sm. Rhizomes

A chemical study on the rhizomes of Hedychium spicatum resulted in the isolation and identification of labdane diterpenes, hedychilactone A (1), and B (2). The chemical structure of known compounds 1 and 2 were elucidated based on extensive spectral data analyses and identified by comparing their experimental spectral data with the reported data in the literature. The isolated labdane diterpenes were evaluated for their pesticide effects like nematicidal activity on Meloidogyne incognita, insecticidal activity on Spodoptera litura, and herbicidal activity on Raphanus raphanistrum seeds. The bioactivity of both compounds was predicted with the help of in silico PASS studies. The proteins/enzymes undertaken for docking studies by AutoDock4.2 to validate nematicidal activity was acetylcholinesterase (PBD ID: IC2O), for insecticidal activity was enzyme carboxylesterase (PDB ID: 1CI8), and for herbicidal activity was acetohydroxyacid synthase (PBD ID: 1YHZ). These compounds could be deduced as a viable alternative source of natural pesticidal agents and pave the way for further studies on its mechanistic approaches and field trials to ascertain its pesticidal studies.

Sublethal effects of chlorantraniliprole on growth, biochemical and molecular parameters in two chironomids, Chironomus kiiensis and Chironomus javanus

Pesticide residues have serious environmental impacts on rice-based ecosystems. In rice fields, Chironomus kiiensis and Chironomus javanus provide alternative food sources to predatory natural enemies of rice insect pests, especially when pests are low. Chlorantraniliprole is a substitute for older classes of insecticides and has been used extensively to control rice pests. To determine the ecological risks of chlorantraniliprole in rice fields, we evaluated its toxic effects on certain growth, biochemical and molecular parameters in these two chironomids. The toxicity tests were performed by exposing third-instar larvae to a range of concentrations of chlorantraniliprole. LC50 values at 24 h, 48 h, and 10 days showed that chlorantraniliprole was more toxic to C. javanus than to C. kiiensis. Chlorantraniliprole significantly prolonged the larval growth duration, inhibited pupation and emergence, and decreased egg numbers of C. kiiensis and C. javanus at sublethal dosages (LC10 = 1.50 mg/L and LC25 = 3.00 mg/L for C. kiiensis; LC10 = 0.25 mg/L and LC25 = 0.50 mg/L for C. javanus). Sublethal exposure to chlorantraniliprole significantly decreased the activity of the detoxification enzymes carboxylesterase (CarE) and glutathione S-transferases (GSTs) in both C. kiiensis and C. javanus. Sublethal exposure to chlorantraniliprole also markedly inhibited the activity of the antioxidant enzyme peroxidase (POD) in C. kiiensis and POD and catalase (CAT) in C. javanus. Expression levels of 12 genes revealed that detoxification and antioxidant abilities were affected by sublethal exposures to chlorantraniliprole. There were significant changes in the expression levels of seven genes (CarE6, CYP9AU1, CYP6FV2, GSTo1, GSTs1, GSTd2, and POD) in C. kiiensis and ten genes (CarE6, CYP9AU1, CYP6FV2, GSTo1, GSTs1, GSTd2, GSTu1, GSTu2, CAT, and POD) in C. javanus. These results provide a comprehensive overview of the differences in chlorantraniliprole toxicity to chironomids, indicating that C. javanus is more susceptible and suitable as an indicator for ecological risk assessment in rice ecosystems.

Fish Carboxyl Esterase Activity as a Possible Hypoxia Biomarker

Physiology and biochemistry adaptations studied in fish are important to comprehend how they endure the challenges of living in aquatic environments. Oxygen variations are a frequent natural and cyclic phenomenon that occurs in the Pantanal and Amazon watersheds and it is the limiting factor to survival of the aerobic species. The growing of pollution in these water basins threaten many species as anthropic hypoxia intensifies the production of reactive oxygen species (ROS) and lipid peroxidation inside the cells, leading organisms to a condition known as oxidative stress. Our research group, studying the effects of the oxygen variations on the carboxyl esterase (CarbE) enzyme of pacu, a typical fish from Pantanal, observed that CarbE activity diminished by half in pacus exposed to 42 hours of hypoxia. Based on this finding, we decided to investigate this phenomenon in other fish species, including hypoxia-tolerant fishes. CarbE activities were determined using p-nitrophenyl acetate (p-NPA) as a substrate. All five species of fish studied had both serum and liver CarbE activity significantly decreased. Pacus had its serum and liver activities decreased by approximately 50% under severe hypoxia.

Cotton Aphid (Hemiptera: Aphididae) Resistance to Afidopyropen in Xinjiang Region, China

Abstract Concentration-mortality response bioassays were conducted in 2021 to define the toxicity of afidopyropen to field populations of cotton aphids, Aphis gossypii Glover (Hemiptera: Aphididae), in Xinjiang region, China. Levels of activity of selected enzymes also were measured. Varying levels of resistance to afidopyropen were detected among aphids from nine major cotton-growing areas in the region. Higher resistance levels were detected in aphids collected from Tumshuk, Alar, and Kurle, with resistance ratios of 4.570, 2.058, and 1.565, respectively. Lower resistance ratios of 0.506, 0.632, and 0.775 were detected in aphids collected from Yinli, Wujiaqu, and Hami, respectively. Biochemical assays showed a highly significant positive correlation of the detoxifying enzymes carboxylesterase and multifunctional oxidase with the level of tolerance, with enzyme activity increasing with the level of tolerance. Glutathione S-transferases and acetylcholinesterase activity in field-collected aphids differed significantly from laboratory colony populations, but their activity did not differ among the field populations. We concluded that cotton aphids in major cotton areas of Xinjiang have not yet developed resistance to afidopyropen and remain sensitive to afidopyropen as indicated by resistance ratios of <5; however, resistance of cotton aphids to afidopyropen in southern Xinjiang is higher than that detected in cotton aphid populations in northern Xinjiang.

A Simple Potentiometric Biosensor based on Carboxylesterase for the Analysis of Aspartame

A potentiometric aspartame biosensor was fabricated by simply depositing the carboxylesterase (CES)-bonded poly(n-butyl acrylate-n-acryloxysuccimide) [CES-poly(nBA-NAS)] microspheres on a Ag/AgCl screen-printed pH selective electrode. The pH transducer was made from non-plasticized polyacrylate membrane containing a hydrogen ionophore and lipophilic salt. The immobilized CES enzyme catalyzed the enzymatic hydrolysis of aspartame to L-aspartic acid (L-Asp), L-phenylalanine and methanol. Potentiometric determination of aspartame concentration was performed by quantifying the hydrogen ion concentration produced from L-Asp. The potentiometric determination of aspartame exhibited good selectivity with near Nernstian response. The sensitivity of the biosensor was closed to the Nernstian value, i.e., 50-52 mV decade-1 with a dynamic linear response range from 10-5 to 10-2 M and detection limit approaching 10-6 M. The aspartame biosensor demonstrated good repeatability and reproducibility with relative standard deviation (RSD) of 1.9% and 1.6%, respectively (n=3). The potentiometric aspartame biosensor was demonstrated to be reliable for determining aspartame content in sweetener samples and was comparable to the conventional high-performance liquid chromatography (HPLC) method for aspartame analysis.

The mechanistic basis for the toxicity difference between juvenile rats and mice following exposure to the agricultural insecticide chlorpyrifos

At high exposure levels, organophosphorus insecticides (OPs) exert their toxicity in mammals through the inhibition of brain acetylcholinesterase (AChE) leading to the accumulation of acetylcholine in cholinergic synapses and hyperactivity of the nervous system. Currently, there is a concern that low-level exposure to OPs induces negative impacts in developing children and the chemical most linked to these issues is chlorpyrifos (CPF). Our laboratory has observed that a difference in the susceptibility to repeated exposure to CPF exists between juvenile mice and rats with respect to the inhibition of brain AChE. The basis for this difference is unknown but differences in the levels of the detoxification mechanisms could play a role. To investigate this, 10-day old rat and mice pups were exposed daily for 7 days to either corn oil or a range of dosages of CPF via oral gavage. Four hours following the last administration of CPF on day 16, brain, blood, and liver were collected. The inhibition of brain AChE activity was higher in juvenile rats as compared to juvenile mice. The levels of activity of the detoxification enzymes and the impact of CPF exposure on their activity were determined in the two species at this age. In blood and liver, the enzyme paraoxonase-1 (PON1) hydrolyzes the active metabolite of CPF (CPF-oxon), and the enzymes carboxylesterase (CES) and cholinesterase (ChE) act as alternative binding sites for CPF-oxon removing it from circulation and providing protection. Both species had similar levels of PON1 activity in the liver and serum. Mice had higher ChE activity in liver and serum than rats but, following CPF exposure, the percentage inhibition was similar between species at an equivalent dosage. Even though rats had slightly higher liver CES activity than mice, the level of inhibition following exposure was higher in rats. In serum, juvenile mice had an 8-fold higher CES activity than rats, and exposure to a CPF dosage that almost eliminated CES activity in rats only resulted in 22% inhibition in mice suggesting that the high serum CES activity in mice as compared to rats is a key component in this species difference. In addition, there was a species difference in the sensitivity of CES to inhibition by CPF-oxon with rats having a lower IC50 in both liver and serum as compared to mice. This greater enzyme sensitivity suggests that saturation of CES would occur more rapidly in juvenile rats than in mice, resulting in more CPF reaching the brain to inhibit AChE in rats.

Effect of microplastics on the activity of carboxylesterase and phosphatase enzymes in Scinax squalirostris tadpoles.

Microplastics (MPs) are critical emerging pollutants around the world. There is a growing interest in the effects of MP ingestion, non-digestion, and toxicity on aquatic organisms. Amphibian tadpoles are the vertebrate group that has received the least attention regarding this issue. The aim of the present study was to determine the ingestion of polyethylene MPs by Scinax squalirostris tadpoles by atomic force microscopy (AFM) and to evaluate the activities of carboxylesterase (CbE, using 4-naphthyl butyrate-NB-, and 1-naphthyl acetate -NA- as substrates) and alkaline phosphatase (ALP) under MP exposure. Enzyme activities were analyzed spectrophotometrically at 2 and 10days of exposure. Tadpoles were exposed to two different treatments during 10days: a negative control (CO, dechlorinated water) and MP (60mg L-1). AFM images of the digestive contents of tadpoles revealed the presence of MPs. After 10days of MP exposure, CbE (NB) activity was significantly higher and CbE (NA) activity was significantly lower in MP treatments than in controls. ALP activity decreased in MP treatments after 2 and 10days of exposure. The detection of MP particles in the intestinal contents and the effects on metabolic enzymes in a common frog species evidenced the potential health risk of MP to aquatic vertebrates. Thus, the differential response in enzymes and substrates demonstrate the need for considering the complex effects of contaminants and nutrients on ecosystems for ecotoxicological risk characterization.

Inhibitory effects of Serjania erecta on the development of Chrysodeixis includens

The soybean looper, Chrysodeixis includens, is a primary soybean pest that reduces crop productivity. This work examined control of C. includens populations with methanolic extract of Serjania erecta, a native Cerrado plant, while minimizing risks to pollinators, natural enemies and the environment. Serjania erecta specimens were collected, identified, and subjected to methanol extraction. Bioassays were performed using newly hatched and second-instar caterpillars and different extract concentrations on the diet surface to obtain IC50 values. Two replicates, containing 10 caterpillars, were established in triplicate. The IC50 values were 4.15 and 6.24 mg of extract mL−1 for first-instar and second-instar caterpillars, respectively. These growth inhibition results informed the extract concentrations assessed in subsequent development inhibition assays, in which the pupal weight was higher under the control than under the treatments. Extract treatments increased the duration of the larval, pupal and total development. The potential of different concentrations of S. erecta extract to inhibit the enzymes carboxylesterases was also evaluated. Carboxylesterases activity decreased by 41.96 and 43.43% at 7.8 and 15.6 μg mL−1 extract, respectively. At 31.3 μg mL−1 extract, enzymatic activity was not detected. Overall, S. erecta leaf methanolic extract showed inhibitory potential against carboxylesterases.

Evaluation of the Safety and Ochratoxin A Degradation Capacity of Pediococcus pentosaceus as a Dietary Probiotic with Molecular Docking Approach and Pharmacokinetic Toxicity Assessment.

The present study evaluated the properties and ochratoxin A (OTA) degradation capacity of the dietary probiotic Pediococcus pentosaceus BalaMMB-P3, isolated from a milk coagulant. The acidic tolerance of the isolate at pH 2–3 was checked with bile salts. No hemolytic activity was noted, which confirmed the nonpathogenicity of the strain. The isolate was tested in vitro for antibiotic susceptibility, enzymatic activity, bile salts hydrolase activity and antifungal activity against Penicillium verrucosum, Fusarium graminearum and Aspergillus ochraceus. A molecular docking-based OTA toxicity assessment was carried out for multitargeted proteins. The 16S rRNA gene-based phylogenetic assessment identified the strain as P. pentosaceus, and was authenticated in GenBank. The carboxylesterase and glutathione s-transferase enzymes showed active and strong interactions with esters and amide bonds, respectively. The compound exhibited carcinogenic and cytotoxicity effects at an LD50 value of 20 mg/kg. Furthermore, the strain showed a potent ability to reduce OTA and suggested the prospects for utilization in nutritional aspects of food.

The Role of Insect Symbiotic Bacteria in Metabolizing Phytochemicals and Agrochemicals.

Simple SummaryTo counter plant chemical defenses and exposure to agrochemicals, herbivorous insects have developed several adaptive strategies to guard against the ingested detrimental substances, including enhancing detoxifying enzyme activities, avoidance behavior, amino acid mutation of target sites, and lower penetration through a thicker cuticle. Insect microbiota play important roles in many aspects of insect biology and physiology. To better understand the role of insect symbiotic bacteria in metabolizing these detrimental substances, we summarize the research progress on the function of insect bacteria in metabolizing phytochemicals and agrochemicals, and describe their future potential application in pest management and protection of beneficial insects.The diversity and high adaptability of insects are heavily associated with their symbiotic microbes, which include bacteria, fungi, viruses, protozoa, and archaea. These microbes play important roles in many aspects of the biology and physiology of insects, such as helping the host insects with food digestion, nutrition absorption, strengthening immunity and confronting plant defenses. To maintain normal development and population reproduction, herbivorous insects have developed strategies to detoxify the substances to which they may be exposed in the living habitat, such as the detoxifying enzymes carboxylesterase, glutathione-S-transferases (GSTs), and cytochrome P450 monooxygenases (CYP450s). Additionally, insect symbiotic bacteria can act as an important factor to modulate the adaptability of insects to the exposed detrimental substances. This review summarizes the current research progress on the role of insect symbiotic bacteria in metabolizing phytochemicals and agrochemicals (insecticides and herbicides). Given the importance of insect microbiota, more functional symbiotic bacteria that modulate the adaptability of insects to the detrimental substances to which they are exposed should be identified, and the underlying mechanisms should also be further studied, facilitating the development of microbial-resource-based pest control approaches or protective methods for beneficial insects.

Unraveling arbuscular mycorrhizal fungi-induced resistance of purple branch rose (Rosa rugosa ‘Zizhi’) to Lymantria dispar based on metabolomics

Currently, people primarily rely on chemical pesticides to prevent and control insect pests, which not only introduce problems with environmental pollution but also produce chemical residues that will make it difficult to guarantee the safety of edible plants. Arbuscular mycorrhizal fungi (AMF) are soil microorganisms that form a symbiotic relationship with plants, which improves plant resistance. This natural phenomenon provides new ideas for eco-friendly biological control. The present study assessed the effects of AMF (Funneliformis mosseae, Rhizophagus intraradices and their combination) inoculation on rose growth and its resistance to gypsy moth larvae (Lymantria dispar) by investigating the growth performance and metabolomics of rose plants and the growth and physiological parameters of gypsy moth larvae. AMF inoculation improved the growth of both the aboveground and belowground parts of the rose plants. The developmental duration of gypsy moth larvae feeding on mycorrhizal rose leaves was prolonged by 4.8–5.8 days. Moreover, feeding on mycorrhizal rose leaves significantly reduced the body weight of the gypsy moth larvae by 9–24%, body length by 4–6%, and the width of head sheath by 11–15%. The superoxide dismutase (SOD) and carboxylesterase (CarE) enzymes in the larvae that fed on the leaves of mycorrhizal rose were significantly higher than those in the control, while the acid phosphatase (ACP) was inhibited (P < 0.05). Peroxidase (POD) and alkaline phosphatase (AKP) in larvae that fed on mycorrhizal rose leaves were significantly higher than those in the control at 5 days. Glutathione S transferases (GSTs) were significantly higher than the control at 3 and 5 days. Metabolomics analyses showed that inoculation with AMF promoted the accumulation of jasmonic acid (JA) and abscisic acid (ABA) in rose leaves. The treatment groups were 1.6- and 3.4-fold higher than those of the control, respectively. The AMF significantly regulated 1,589 metabolites in rose leaves. The AMF promoted the accumulation of flavonoids and inhibited the accumulation of glycosides in rose leaves. A total of 19 metabolites that significantly correlated with the growth and development of gypsy moths were screened out. Among ABA, glycerophosphocholine, 1-(sn-glycero-3-phospho)-1D-myo-inositol, (-)-catechin, N-acetyl-D-glucosamine 6-phosphate, 3-(4-hydroxyphenyl)-1-propanol and 3-trans-caffeoyltormentic acid were significantly different from those of the control. These compounds may play a role in the resistance of rose to gypsy moth induced by AMF. In conclusion, AMF inoculation improved the resistance of R. rugosa to L. dispar through modifications on the metabolism and physiology. These findings highlight the interaction of “mycorrhizal-plant-insect”. It provides a reference for the application of AMF to control plant pest.

Resistance in Spodoptera litura (F.) to Insecticides and Detoxification Enzymes

Standard leaf dip bioassay experiments were conducted to study the insecticide resistance in Spodoptera litura (F.) in soybean. Chlorantraniliprole was the most toxic while, cypermethrin and organophosphates (profenophos and triazophos) recorded least toxicity against S. litura populations. Field population of S. litura showed the highest resistance to cypermethrin with resistance ratios (RRs) ranging from 244 to 376. The field populations collected from three locations of Southern Rajasthan exhibited low resistance to newer insecticides, chlorantraniliprole and spinetoram with Lab-SS strain. All the enzyme activities were significantly higher in the whole larval extracts and midgut extracts of S. litura larvae when compared to the Lab-SS strain. Analysis of the carboxyl esterase enzyme in the native PAGE assay revealed that the electrophoretic profiles showed presence of different band regions (EST 1 to EST 7) with esterase activity in the field populations along with the laboratory susceptible strain. The susceptibility of three field populations to different group of insecticides along with the profile of detoxification enzymes indicate the need to formulate a region-specific insecticide resistance management (IRM).

Liposomal irinotecan (Onivyde): Exemplifying the benefits of nanotherapeutic drugs.

Irinotecan is a topoisomerase inhibitor, widely used in treatment of malignancies including pancreatic ductal adenocarcinoma (PDAC) as part of the FOLFIRINOX regimen prescribed as a first‐line treatment in several countries. However, irinotecan has not been successfully introduced as a second‐line treatment for pancreatic cancer and few randomized clinical studies have evaluated its added value. Efficacy of liposomal irinotecan (nal‐IRI) combined with 5‐fluorouracil and leucovorin (5‐FU/LV) was reported in the phase III NAPOLI‐1 trial in metastatic PDAC following failure of gemcitabine‐based therapy. Several features of nal‐IRI pharmacokinetics (PK) could result in better outcomes versus nonliposomal irinotecan. Irinotecan is a prodrug that is converted to active SN‐38 by carboxylesterase enzymes and inactivated by cytochrome P450 3A4/3A5. SN‐38 is inactivated by UGT1A1 enzymes. Individual variations in their expression and activity could influence enhanced localized irinotecan activity and toxicity. Liposomal irinotecan exploits the enhanced permeability and retention effect in cancer, accumulating in tumor tissues. Liposomal irinotecan also has a longer half‐life and higher area under the concentration‐time curve (0–∞) than nonliposomal irinotecan, as the liposomal formulation protects cargo from premature metabolism in the plasma. This results in irinotecan activation in tumor tissue, leading to enhanced cytotoxicity. Importantly, despite the longer exposure, overall toxicity for nal‐IRI is no worse than nonliposomal irinotecan. Liposomal irinotecan exemplifies how liposomal encapsulation of a chemotherapeutic agent can alter its PK properties, improving clinical outcomes for patients. Liposomal irinotecan is currently under investigation in other malignancies including biliary tract cancer (amongst other gastrointestinal cancers), brain tumors, and small‐cell lung cancer.

Evaluation of toxic effect of insecticide, herbicide and fungicide as single and mixture on Gammarus lacustris using biochemical markers

ABSTRACT In the present study, the acute toxic effects of single, double, and triple mixtures of imidacloprid, azoxystrobin, and glyphosate on Gammarus lacustris were investigated. Initially, the 96-hour median lethal dose (LC50) values for imidacloprid, azoxystrobin, and glyphosate were determined as 157, 1.92, and 4.33 mg L−1, respectively. The animals were exposed to one-fiftieth of the 96-h LC50 concentration for 24 or 96 h. The activity of glutathione S-transferase (GST), glutathione reductase, acetylcholinesterase, carboxylesterase (CaE), and superoxide dismutase enzymes as biological markers were examined to determine the non-lethal effects of the pesticides tested. Significant changes were observed in GST and CaE activities especially in detoxification compared to control and between different applications. The results of this study show the potential for increased toxicity in organisms exposed to pesticide mixtures.

Realistic exposure to fipronil, 2,4-D, vinasse and their mixtures impair larval amphibian physiology

Expansion of sugarcane crops may have contributed to the increased contamination of native habitats in Brazil. Several species of amphibians inhabit ponds formed in flooded farmlands, where pesticide concentrations are usually high. This study evaluated the ecotoxicological effects of the sugarcane pesticides fipronil and 2,4-D, as well as the fertilizer vinasse (isolated and mixed), on physiological responses of Leptodactylus fuscus and Lithobates catesbeianus tadpoles. In situ assays were conducted in mesocosms with concentrations based on the doses recommended by the manufacturer. Vinasse (1.3% dilution) caused 100% tadpoles’ mortality immediately after its application. Fipronil and/or 2,4-D altered antioxidant and biotransformation responses, induced neurotoxicity and changed lipid contents in tadpoles. A multivariate approach indicated that the mixture of pesticides induced most of the sublethal effects in both tadpole species, in addition to the isolated fipronil in L. fuscus. Fipronil alone increased glucose-6-phosphate dehydrogenase (G6PDH) activity, decreased acetylcholinesterase (AChE) and total lipid contents, and altered some individual lipid classes (e.g., free fatty acids and acetone-mobile polar lipids) in L. fuscus. The interaction between fipronil and 2,4-D in this species were more evident for lipid contents, although enzymatic alterations in G6PDH, AChE and glutathione-S-transferase (GST) were also observed. In L. catesbeianus, the mixture of pesticides reduced triglycerides and total lipids, as well as increased GST and decreased AChE activities. The detoxifying enzyme carboxylesterase was reduced by 2,4-D (alone or in mixture) in both species. Isolated pesticides also modulated specific lipid classes, suggesting their disruptive action on energy metabolism of tadpoles. Our study showed that fipronil, 2,4-D, and vinasse, individually or mixed, can be harmful to amphibians during their larval phase, causing mortality or impairing their functional responses.