Detoxification Function Research Articles

Transgenic expression of SeCYP9A186 and PxFMO2 confers resistance to emamectin benzoate in Plutella xylostella.

The overexpression of metabolic enzymes constitutes a crucial mechanism for insects to detoxify xenobiotics and metabolic pesticides. A flavin-containing monooxygenase gene (PxFMO2) from Plutella xylostella and a P450 gene (SeCYP9A186, F116V mutant allele) from Spodoptera exigua have been reported to be involved in insecticide resistance. In this study, we aim to utilize transgenic technology to validate their in vivo detoxification functions in Plutella xylostella. We established two transgenic strains of Plutella xylostella with expressing an endogenous PxFMO2 gene from Plutella xylostella and an exogenous SeCYP9A186 gene from S. exigua, respectively. Bioassays demonstrated that the transgenic Plutella xylostella strain (IPP-FMO2) expressing PxFMO2 exhibited a 12-fold resistance to emamectin benzoate and a 6.4-fold resistance to chlorantraniliprole compared to the background strain (IPP-S). In contrast, the transgenic Plutella xylostella strain (IPP-9A186) expressing SeCYP9A186 displayed a 235-fold resistance to emamectin benzoate and a 115-fold resistance to abamectin. Moreover, resistance to emamectin benzoate in the IPP-9A186 strain of Plutella xylostella was inherited as an incompletely dominant trait and was genetically linked to the transgene locus. Our results not only elucidated the in vivo contribution of the PxFMO2 and SeCYP9A186 to the insecticide resistance phenotype in Plutella xylostella, but also provided a genetic engineering toolkit to manipulate resistance pathways. These insights and methodologies could further aid in developing sustainable pest management strategies in Plutella xylostella. © 2024 Society of Chemical Industry.

Integrating morphology, physiology, and computer simulation to reveal the toxicity mechanism of eco-friendly rosin-based pesticides

To mitigate the impact of traditional chemical pesticides on environment, and achieve sustainable crop protection, 24 eco-friendly rosin-based sulfonamide derivatives were synthesized and developed. The in vitro activity assessment showed that compound 4X (Co. 4X) exhibited excellent fungicidal activity against V. mali (EC50 = 1.106 μg/mL), marginally surpassing the positive control carbendazim (EC50 = 1.353 μg/mL). In vivo investigations suggested that Co. 4X exhibited moderate efficacy in mitigating V. mali infection in both apple trees and apples. Physiological assessments revealed that Co. 4X induced severe ultrastructural damage to the mycelium, heightened cell membrane permeability, and inhibited SDH protein activity. Subsequent biosafety evaluations affirmed the environment-friendly of Co. 4X on Zebrafish (LC50(96h) = 25.176 μg/mL). Toxicological research revealed that Co. 4X caused damage to the cells of Zebrafish gills, liver, and intestines, resulting in impaired respiratory, detoxification, digestion, and absorption functions of Zebrafish. In summary, the findings of this study contribute to a deeper understanding of the toxicity mechanisms of novel pesticides, decreasing environmental risks caused by traditional chemical pesticides, and improving the effective management of novel pesticide applications.

Significance of the liver arginase activity in the processes of detoxification, formation of thyroid status and dyslipidemia in rats with experimental peritonitis

Peritonitis is a surgical and general pathological problem, the relevance of which does not decrease despite the achievements of modern clinical and experimental medicine. Peritonitis, being one of the most severe complications of various diseases and injuries to the abdominal organs, triggers a complex cascade of pathogenetic reactions with disruption of vital processes. The mortality rate for peritonitis is 20‒30 %, and in its most severe forms it reaches 50–80 %. High mortality is caused by endotoxemia, multiple organ failure and sepsis. To date, sufficient data have been accumulated, indicating the importance of liver arginase in the processes of detoxification and vital activity of the body in normal conditions and in pathology. There were reasons to believe that its activity would be significant in the pathogenesis of peritonitis. However, the elucidation of significance of the liver arginase activity in detoxification processes and in the pathogenesis of septic conditions and peritonitis in particular has been still little studied.The purpose of the study was to determine the significance of the liver arginase activity in the processes of detoxification, the formation of thyroid status and dyslipidemia in rats with experimental peritonitis (CLP-model).In experiments on rats, it was found that liver arginase and nitrogen monoxide are involved in changes in the content of total cholesterol in the liver and lipoproteins in the blood serum, the level of iodine-containing thyroid hormones in the blood plasma and body temperature during CLP-peritonitis. Developing CLP-peritonitis in the conditions of liver argina- se depression by Nω-hydroxy-nor-L-arginine at a dose of 10 mg/kg is accompanied by a more pronounced inhibition of the detoxification function of the liver, worsening changes in the content of total cholesterol in the liver and lipoproteins in the blood serum, and the level of iodine-containing hormones in the blood plasma and contributes to the development of secondary dislipoproteinemia.

Isotschimgine promotes lifespan, healthspan and neuroprotection of Caenorhabditis elegans via the activation of nuclear hormone receptors.

Isotschimgine (ITG) is a bornane-type monoterpenoid derivative naturally occurring in genus Ferula plants and propolis. Its effects on aging and the underlying mechanisms are not yet well understood. This study employed Caenorhabditis elegans (C. elegans) as a model organism to evaluate the potential of ITG in extending lifespan, enhancing healthspan, and promoting neuroprotection, while exploring the underlying mechanisms involved. The results showed that ITG extended the lifespan and healthspan of C. elegans, significantly enhanced stress resistance and detoxification functions. Studies on mutants and qPCR data indicated that ITG-mediated lifespan extension was modulated by the insulin/IGF-1 signaling pathway and nuclear hormone receptors. Furthermore, ITG markedly increased stress-responsive genes, including daf-16 and its downstream genes sod-3 and hsp-16.2, as well as NHR downstream detoxification-related genes cyp35a1, cyp35b3, cyp35c1, gst-4, pgp-3 and pgp-13. Additionally, ITG alleviated β-amyloid-induced paralysis and behavioral dysfunction in transgenic C. elegans strains. The neuroprotective efficacy of ITG was weakened by RNAi knockdown of nuclear hormone receptors daf-12 and nhr-8. Overall, our study identifies ITG as a potential compound for promoting longevity and neuroprotection, mediated through nuclear hormone receptors.

Extreme heat event influences the toxic impacts of nano-TiO2 with different crystal structures in mussel Mytilus coruscus

The wide use of nano‑titanium dioxide (nano-TiO2) and its ubiquitous emission into aquatic environments are threatening environmental health. Ambient temperature can affect the aggregation state of nano-TiO2 in seawater, thus influencing the intake and physiological effects on marine species. We studied the physiological effects of mixed nano-TiO2 (a mixture of anatase and rutile crystals with an average particle size of 25 nm, P25) on mussels. Subsequently, we investigated the oxidative stress, immunotoxicity, neurotoxicity, and detoxification in Mytilus coruscus exposed to two different crystal structures of nano-TiO2 (anatase and rutile) at 100 μg/L concentration under marine heatwaves (MHWs, 28 °C). MHWs and nano-TiO2 exposure induced neurotoxicity and immune damage and caused dysregulation of redox balance in the gills. Moreover, MHWs exposure disturbed the glutathione system and detoxification function of mussels, resulting in enhanced toxicity of nano-TiO2 under co-exposure. Anatase exposure significantly impaired the antioxidant system and downregulated the relative expression of antioxidant-related genes (Nrf2 and Bcl-2), HSP-90, and immune parameters under MHWs, while producing higher ROS levels compared to rutile. Based on integrated biomarker response (IBR), mussels co-exposed to anatase and MHW showed the highest value (19.29). However, there was no significant difference in bioaccumulation of titanium between anatase (6.07 ± 0.47 μg/g) and rutile (5.3 ± 0.44 μg/g) exposures under MHWs. These results indicate that MHWs would elevate the potential hazard of nanoparticles to marine organisms.

Identification and functional validation of P450 genes associated with pyrethroid resistance in the malaria vector Anopheles sinensis (Diptera Culicidae)

Cytochrome P450 monooxygenases (P450s), a multifunctional protein superfamily, are one of three major classes of detoxification enzymes. However, the diversity and functions of P450 genes from pyrethroid-resistant populations of Anopheles sinensis have not been fully explored. In this study, P450 genes associated with pyrethroid resistance were systematically screened using RNA-seq in three field pyrethroid-resistant populations (AH-FR, CQ-FR, YN-FR) and one laboratory resistant strain (WX-LR) at developmental stages, tissues, and post blood-meal in comparison to the laboratory susceptible strain (WX-LS) in An. sinensis. Importantly, the expression of significantly upregulated P450s was verified using RT-qPCR, and the function of selected P450s in pyrethroid detoxification was determined with RNA interference using four laboratory pyrethroid-resistant strains (WX-LR, AH-LR, CQ-LR, YN-LR). Sixteen P450 genes were significantly upregulated in at least one field-resistant population, and 44 were significantly upregulated in different developmental stages, tissues or post blood-meal. A total of 19 P450s were selected to verify their association with pyrethroid resistance, and four of them (AsCYP6P3v1, AsCYP6P3v2, AsCYP9J10, and AsCYP9K1) demonstrated significant upregulation in laboratory pyrethroid-resistant strains using RT-qPCR. Knockdown of these four genes all significantly reduced pyrethroid resistance and increased the mortality by 57.19% (AsCYP6P3v1 and AsCYP6P3v2 knockdown group), 38.39% (AsCYP9K1 knockdown group) and 48.87% (AsCYP9J10 knockdown group) in An. sinensis by RNAi, which determined the pyrethroid detoxification function of these four genes. This study revealed the diversity of P450 genes and provided functional evidence for four P450s in pyrethroid detoxification in An. sinensis for the first time, which increases our understanding of the pyrethroid resistance mechanism, and is of potential value for pyrethroid resistance detection and surveillance.

Root Endophytic Microorganisms Contribute to the Attribute of Full-Year Shooting in Woody Bamboo Cephalostachyum pingbianense.

Cephalostachyum pingbianense (Hsueh & Y.M. Yang ex Yi et al.) D.Z. Li & H.Q. Yang is unique among bamboo species for its ability to produce bamboo shoots in all seasons under natural conditions. Apart from the physiological mechanism, information regarding the effects of endophytic microorganisms on this full-year shooting characteristic is limited. We hypothesize that root endophytic microorganisms will have a positive impact on the full-year bamboo shooting characteristic of C. pingbianense by increasing the availability or supply of nutrients. To identify the seasonal variations in the root endophytic bacterial and fungal communities of C. pingbianense, and to assess their correlation with bamboo shoot productivity, the roots of C. pingbianense were selected as research materials, and the 16S rRNA and ITS rDNA genes of root endophytic microorganisms were sequenced using the Illumina platform. Following this sequencing, raw sequencing reads were processed, and OTUs were annotated. Alpha and beta diversity, microbial composition, and functional predictions were analyzed, with correlations to bamboo shoot numbers assessed. The results showed that seasonal changes significantly affected the community diversity and structure of root endophytic microbes of C. pingbianense. Bacterial communities in root samples from all seasons contained more nitrogen-fixing microorganisms, with members of the Burkholderiales and Rhizobiales predominating. The relative abundances of ectomycorrhizal and arbuscular mycorrhizal fungi in the autumn sample were significantly higher than in other seasons. Correlation analysis revealed that the bamboo shoot productivity was significantly and positively correlated with bacterial functions of nitrogen fixation, arsenate detoxification, and ureolysis, as well as with symbiotrophic fungi, ectomycorrhizal fungi, and arbuscular mycorrhizal fungi. At the genus level, the bacterial genus Herbaspirillum and the fungal genera Russula, unclassified_f_Acaulosporaceae, and unclassified_f_Glomeraceae were found to have a significant positive correlation with bamboo shoot number. Our study provides an ecological perspective for understanding the highly productive attribute of C. pingbianense and offers new insights into the forest management of woody bamboos.

197 Hepatic mRNA expression of innate and adaptive immune genes in beef steers with divergent residual body weight gain

Abstract Immune function has a pivotal role in dictating the overall health and productivity of cattle. In a proficient immune system, the liver assumes an integral function in detoxification and metabolic processes and contributes substantially to overall production and immunity. In this study, we evaluated the hepatic mRNA expression of genes involved in innate and adaptive immunity in crossbred beef steers with positive or negative residual body weight (BW) gain (RADG). Positive-RADG beef steers (n = 8; RADG = 0.76 kg/d) and negative-RADG beef steers (n = 8; RADG = -0.64 kg/d) were identified from a group of 108 growing crossbred beef steers (average BW = 286 ± 380kg) after a 56-d performance testing period. At the end of the 56-d period, liver tissue samples were collected from the beef steers for RNA extraction and cDNA synthesis. The mRNA expression of 84 genes involved in innate and adaptive immunity were analyzed using pathway-focused PCR-based arrays. The mRNA expression of genes with false discovery rate-adjusted P-values (FDR) ≤ 0.05 and absolute fold change (FC) ≥ 1.2 were determined to be differentially expressed. Out of the 84 genes analyzed, four genes [interleukin (IL)-2, MYD88, CD-80, NFkB-1] were differentially expressed and were all upregulated in positive compared with negative-RADG beef steers. Gene ontology pathway enrichment analysis of the differentially expressed genes revealed that pathways related to positive regulation of IL-17 production, cellular response to lipopolysaccharide, and positive regulation of lymphocyte activation were enriched in the positive-RADG steers (P ≤ 0.05). In conclusion, our study revealed that positive-RADG beef steers had increased expression of genes and pathways involved in pathogen recognition and initiation of responses to effectively fight off infections while mitigating excess inflammatory reactions compared with negative-RADG steers, which might explain, in part, the improved feed efficiency of these beef animals

Dental Sedation and General Anesthesia Considerations for Patients Posthepatic Transplantation.

Hepatic compromise poses significant impacts upon the care of patients undergoing routine dental treatment. When sedation or general anesthesia is planned for dental treatment or oral and maxillofacial surgery, an understanding of basic liver function and clinical evaluation can assist in adapting treatment modifications for patients with limited function due to previous disease and resultant organ transplantation efforts. Beginning with a basic overview of hepatic physiology, this review will outline the specific functions of digestion, metabolism, synthesis, and detoxification involving the liver. Specific clinical considerations will be reviewed regarding comorbidities that develop prior to and after liver transplantation that often impact a patient's suitability for ambulatory and office-based care. Lastly, choices in both local anesthetics, sedative medications, general anesthetics, and postoperative analgesics utilized in dental treatment will be discussed.

NMDA Receptors and Indices of Energy Metabolism in Erythrocytes: Missing Link to the Assessment of Efficiency of Oxygen Transport in Hepatic Encephalopathy.

Hepatic encephalopathy (HE) is a neuropsychiatric syndrome that develops in patients with severe liver dysfunction and/or portocaval shunting. Despite more than a century of research into the relationship between liver damage and development of encephalopathy, pathogenetic mechanisms of hepatic encephalopathy have not yet been fully elucidated. It is generally recognized, however, that the main trigger of neurologic complications in hepatic encephalopathy is the neurotoxin ammonia/ammonium, concentration of which in the blood increases to toxic levels (hyperammonemia), when detoxification function of the liver is impaired. Freely penetrating into brain cells and affecting NMDA-receptor-mediated signaling, ammonia triggers a pathological cascade leading to the sharp inhibition of aerobic glucose metabolism, oxidative stress, brain hypoperfusion, nerve cell damage, and formation of neurological deficits. Brain hypoperfusion, in turn, could be due to the impaired oxygen transport function of erythrocytes, because of the disturbed energy metabolism that occurs in the membranes and inside erythrocytes and controls affinity of hemoglobin for oxygen, which determines the degree of oxygenation of blood and tissues. In our recent study, this causal relationship was confirmed and novel ammonium-induced pro-oxidant effect mediated by excessive activation of NMDA receptors leading to impaired oxygen transport function of erythrocytes was revealed. For a more complete evaluation of "erythrocytic" factors that diminish brain oxygenation and lead to encephalopathy, in this study, activity of the enzymes and concentration of metabolites of glycolysis and Rapoport-Lubering shunt, as well as morphological characteristics of erythrocytes from the rats with acute hyperammoniemia were determined. To elucidate the role of NMDA receptors in the above processes, MK-801, a non-competitive receptor antagonist, was used. Based on the obtained results it can be concluded that it is necessary to consider ammonium-induced morphofunctional disorders of erythrocytes and hemoglobinemia which can occur as a result of alterations in highly integrated networks of metabolic pathways may act as an additional systemic "erythrocytic" pathogenetic factor to prevent the onset and progression of cerebral hypoperfusion in hepatic encephalopathy accompanied by hyperammonemia.

Combined toxicity of abamectin and carbendazim on enzymatic and transcriptional levels in the soil-earthworm microcosm.

To reveal the toxicological mechanisms of pesticide mixtures on soil organisms, this study concentrated on evaluating enzymatic activity and gene expression changes in the earthworm Eisenia fetida (Savigny 1826). Despite being frequently exposed to multiple pesticides, including the common combination of abamectin (ABA) and carbendazim (CAR), environmental organisms have primarily been studied for the effects of individual pesticides. Acute toxicity results exhibited that the combination of ABA and CAR caused a synergistic impact on E. fetida. The levels of MDA, ROS, T-SOD, and caspase3 demonstrated a significant increase across most individual and combined groups, indicating the induction of oxidative stress and cell death. Additionally, the expression of three genes (hsp70, gst, and crt) exhibited a significant decrease following exposure to individual pesticides and their combinations, pointing toward cellular damage and impaired detoxification function. In contrast, a noteworthy increase in ann expression was observed after exposure to both individual pesticides and their mixtures, suggesting the stimulation of reproductive capacity in E. fetida. The present findings contributed to a more comprehensive understanding of the potential toxicity mechanisms of the ABA and CAR mixture, specifically on oxidative stress, cell death, detoxification dysfunction, and reproductive capacity in earthworms. Collectively, these data offered valuable toxicological insights into the combined effects of pesticides on soil organisms, enhancing our understanding of the underlying risks associated with the coexistence of different pesticides in natural soil environments.

Insuficiencia hepática aguda por adenovirus tratada con inmunoglobulina intravenosa

Background: Acute liver failure in pediatric age is a serious multisystem disease, characterized by a failure of the synthesis and detoxification function of the liver. Among the etiologies, viral infection should be investigated. Treatment is supportive and some cases require liver transplantation. Case report: A 2-year-old girl was admitted for acute liver failure. The PCR viral panel was positive for Adenovirus 41 and IgG antibodies to SARS-CoV-2 were also found. Supportive treatment was started without improvement, so intravenous immunoglobulin was administered, with resolution of the liver failure. Conclusion: Immunoglobulin has immunomodulatory mechanisms in children with severe acute hepatitis of infectious etiology, so in some cases, its administration can be considered as adjuvant therapy. Keywords: Acute liver failure, Hepatitis, Adenovirus, Immunoglobulin.

Insuficiencia hepática aguda por adenovirus tratada con inmunoglobulina intravenosa

Background: Acute liver failure in pediatric age is a serious multisystem disease, characterized by a failure of the synthesis and detoxification function of the liver. Among the etiologies, viral infection should be investigated. Treatment is supportive and some cases require liver transplantation. Case report: A 2-year-old girl was admitted for acute liver failure. The PCR viral panel was positive for Adenovirus 41 and IgG antibodies to SARS-CoV-2 were also found. Supportive treatment was started without improvement, so intravenous immunoglobulin was administered, with resolution of the liver failure. Conclusion: Immunoglobulin has immunomodulatory mechanisms in children with severe acute hepatitis of infectious etiology, so in some cases, its administration can be considered as adjuvant therapy. Keywords: Acute liver failure, Hepatitis, Adenovirus, Immunoglobulin.

Isolation and characterization of a hepatoprotective polysaccharide from Lonicera caerulea L. var. edulis Turcz. ex Herd. fruit against APAP-induced acute liver injury mice

The structure and bioactivities of a novel polysaccharide from Lonicera caerulea L. var. edulis Turcz. ex Herd. fruit (THP-3) were investigated. The crude polysaccharides of Turcz. ex Herd. (THP) were extracted by hot water extraction. After purification, the chemical structure of polysaccharides was identified. Then, a mouse model of acute drug-induced liver injury was constructed using 4-acetamidophenol (APAP) and pretreated with THP. The number-average molecular weight of THP-3 was 48.89 kDa and the mass average molar mass was 97.87 kDa. THP-3 was mainly composed of arabinose (42.54 %), glucose (27.62 %), galacturonic acid and galactose (29.84 %). The main linkage types of THP-3 were 1-linked Araf, 1,4-linked Glcp, and 1,3,6-linked Galp. In addition, after THP treatment, serum Alanine aminotransferase (ALT), Aspartate aminotransferase (AST) and γ-glutamyl transpeptidase (γGT) in AILI mice were successfully down-regulated. The results showed that THP could prevent the characteristic morphological changes of hepatic lobular injury and lipid depletion caused by APAP, reduced the level of oxidative damage in mice, increased the expression of APAP-induced hypolipidemia and related inflammatory indicators, and improved the detoxification function of liver. In general, the newly extracted THP polysaccharide has a good liver protection effect and is an ideal natural medicine for the treatment of liver diseases.

Cadmium and ketoprofen accumulation influences aquatic ecosystem demonstrated using in-vivo zebrafish model

The growing concern about pollution and toxicity in aquatic as well as terrestrial organisms is predominantly caused due to waterborne exposure and poses a risk to environmental systems and human health. This study addresses the co-toxic effects of cadmium (Cd) and ketoprofen (KPF), representing heavy metal and pharmaceutical discharge pollutants, respectively, in aquatic ecosystems. A 96-h acute toxicity assessment was conducted using zebrafish embryos. The results indicated that high dosages of KPF (10, 15, and 100 µg/mL) and Cd (10 and 15 µg/mL) reduced survivability and caused concentration-dependent deformities such as scoliosis and yolk sac edema. These findings highlight the potential defects in development and metabolism, as evidenced by hemolysis tests demonstrating dose-dependent effects on blood cell integrity. Furthermore, this study employs adult zebrafish for a 42-day chronic exposure to Cd and KPF (10 and 100 µg/L) alone or combined (10 + 10 and 100 + 100 µg/L) to assess organ-specific Cd and KPF accumulation in tissue samples. Organ-specific accumulation patterns underscore complex interactions impacting respiratory, metabolic, and detoxification functions. Prolonged exposure induces reactive oxygen species formation, compromising antioxidant defense systems. Histological examinations reveal structural changes in gills, gastrointestinal, kidney, and liver tissues, suggesting impairments in respiratory, osmoregulatory, nutritional, and immune functions. This study emphasizes the importance of conducting extensive research on co-toxic effects to assist with environmental risk assessments and safeguard human health and aquatic ecosystems.

First Application of a Mixed Porcine-Human Repopulated Bioengineered Liver in a Preclinical Model of Post-Resection Liver Failure.

In this study, a mixed porcine-human bioengineered liver (MPH-BEL) was used in a preclinical setup of extracorporeal liver support devices as a treatment for a model of post-resection liver failure (PRLF). The potential for human clinical application is further illustrated by comparing the functional capacity of MPH-BEL grafts as assessed using this porcine PRLF model with fully human (FH-BEL) grafts which were perfused and assessed in vitro. BEL grafts were produced by reseeding liver scaffolds with HUVEC and primary porcine hepatocytes (MPH-BEL) or primary human hepatocytes (FH-BEL). PRLF was induced by performing an 85% liver resection in domestic white pigs and randomized into the following three groups 24 h after resection: standard medical therapy (SMT) alone, SMT + extracorporeal circuit (ECC), and SMT + MPH-BEL. The detoxification and metabolic functions of the MPH-BEL grafts were compared to FH-BEL grafts which were perfused in vitro. During the 24 h treatment interval, INR values normalized within 18 h in the MPH-BEL therapy group and urea synthesis increased as compared to the SMT and SMT + ECC control groups. The MPH-BEL treatment was associated with more rapid decline in hematocrit and platelet count compared to both control groups. Histological analysis demonstrated platelet sequestration in the MPH-BEL grafts, possibly related to immune activation. Significantly higher rates of ammonia clearance and metabolic function were observed in the FH-BEL grafts perfused in vitro than in the MPH-BEL grafts. The MPH-BEL treatment was associated with improved markers of liver function in PRLF. Further improvement in liver function in the BEL grafts was observed by seeding the biomatrix with human hepatocytes. Methods to reduce platelet sequestration within BEL grafts is an area of ongoing research.

Multiple UDP glycosyltransferases modulate benzimidazole drug sensitivity in the nematode Caenorhabditis elegans in an additive manner

Xenobiotic biotransformation is an important modulator of anthelmintic drug potency and a potential mechanism of anthelmintic resistance. Both the free-living nematode Caenorhabditis elegans and the ruminant parasite Haemonchus contortus biotransform benzimidazole drugs by glucose conjugation, likely catalysed by UDP-glycosyltransferase (UGT) enzymes. To identify C. elegans genes involved in benzimidazole drug detoxification, we first used a comparative phylogenetic analysis of UGTs from humans, C. elegans and H. contortus, combined with available RNAseq datasets to identify which of the 63 C. elegans ugt genes are most likely to be involved in benzimidazole drug biotransformation. RNA interference knockdown of 15 prioritized C. elegans genes identified those that sensitized animals to the benzimidazole derivative albendazole (ABZ). Genetic mutations subsequently revealed that loss of ugt-9 and ugt-11 had the strongest effects. The “ugt-9 cluster” includes these genes, together with six other closely related ugts. A CRISPR-Cas-9 deletion that removed seven of the eight ugt-9 cluster genes had greater ABZ sensitivity than the single largest-effect mutation. Furthermore, a double mutant of ugt-22 (which is not a member of the ugt-9 cluster) with the ugt-9 cluster deletion further increased ABZ sensitivity. This additivity of mutant phenotypes suggest that ugt genes act in parallel, which could have several, not mutually exclusive, explanations. ugt mutations have different effects with different benzimidazole derivatives, suggesting that enzymes with different specificities could together more efficiently detoxify drugs. Expression patterns of ugt-9, ugt-11 and ugt-22 gfp reporters differ and so likely act in different tissues which may, at least in part, explain their additive effects on drug potency. Overexpression of ugt-9 alone was sufficient to confer partial ABZ resistance, indicating increasing total UGT activity protects animals. In summary, our results suggest that the multiple UGT enzymes have overlapping but not completely redundant functions in benzimidazole drug detoxification and may represent “druggable” targets to improve benzimidazole drug potency.

Evaluation of the Clinical Effectiveness of the Drug Laennec in Complex Rehabilitation of Patients with Post-COVID Syndrome: a Randomized Trial

INTRODUCTION. The development of post-COVID syndrome is accompanied by an increase in markers of systemic inflammation, a violation of the detoxification function of the liver, caused by both direct viral damage to hepatocytes and an increased iatrogenic load on the hepatobiliary system due to polypharmacy. AIM. Studying the use of the drug Laennec in the rehabilitation period as a hepatoprotector and immunomodulator in patients with post-COVID syndrome. MATERIALS AND METHODS. The study included 40 patients with post-COVID syndrome aged 30 to 60 years, with steatosis and steatohepatitis (increased levels of liver transaminases). The clinical effectiveness of Laennec was assessed based on the dynamics of pro-inflammatory biomarkers, liver metabolism indicators, as well as functional tests and psychological questionnaires. RESULTS. The inclusion of Laennec in the rehabilitation program for patients with post-COVID syndrome leads to a more pronounced improvement in lipid metabolism parameters, correction of liver metabolism parameters, a decrease in the level of pro-inflammatory biomarkers, and an improvement in the quality of life of patients than in the control group. DISCUSSION. Significantly significant positive dynamics in the severity of complaints of general weakness and sleep disturbance, as well as indicators of memory impairment and impaired concentration in the main group could indicate the achievement of anabolic, neurotrophic and bioenergetic effects of the Laennec drug. A significantly significant decrease in the level of liver transaminases in the main group indicates the hepatoprotective effect of the drug Laennec, due to the amino acids, vitamins and microelements included in the drug, which support the reactions of phases 1 and 2 of liver detoxification. Normalization of elevated pro-inflammatory markers (ferritin, IL-6, CRP) in patients of the main group may indicate the presence of anti-inflammatory and immunoregulatory effects of the Laennec drug. CONCLUSIONS. A course prescription of the drug Laennec for the purpose of correcting pro-inflammatory markers and hepatoprotection can be recommended for use in complex non-drug rehabilitation to increase its clinical effectiveness, as well as improve subjective indicators of the quality of life of patients with post-COVID syndrome.

Urinary proteomics for noninvasive monitoring of biomarkers of chronic mountain sickness in a young adult population using data-independent acquisition (DIA)-based mass spectrometry

Different populations exhibit varying pathophysiological responses to plateau environments. Therefore, it is crucial to identify molecular markers in body fluids with high specificity and sensitivity to aid in determination. Proteomics offers a fresh perspective for investigating protein changes linked to diseases. We utilize urine as a specific biomarker for early chronic mountain sickness (CMS) detection, as it is a simple-to-collect biological fluid. We collected urine samples from three groups: plains health, plateau health and CMS. Using DIA's proteomic approach, we found differentially expressed proteins between these groups, which will be used as a basis for future studies to identify protein markers. Compared with the healthy plain population, 660 altering proteins were identified in plateau health, which performed the resistance to altitude response function by boosting substance metabolism and reducing immune stress function. Compared to the healthy plateau population, the CMS group had 140 different proteins identified, out of which 8 were potential biomarkers for CMS. Our study has suggested that CMS may be closely related to increased thyroid hormone levels, oxidative damage to the mitochondria, impaired cell detoxification function and inhibited hydrolase activity. SignificanceOur team has compiled a comprehensive dataset of urine proteomics for AMS disease. We successfully identified differentially expressed proteins between healthy and AMS groups using the DIA proteomic approach. We discovered that 660 proteins were altered in plateau health compared to the healthy plain population, resulting in a heightened resistance to altitude response function by boosting substance metabolism and reducing immune stress function. Additionally, we pinpointed 140 different proteins in the AMS group compared to the healthy plateau population, with 8 showing potential as biomarkers for AMS. Our findings suggest that the onset of AMS may be closely linked to increased thyroid hormone levels, oxidative damage to the mitochondria, impaired cell detoxification function and inhibited hydrolase activity.

Adaptogenic correction of free radical processes in liver tissue under the exposure to coal-rock dust on the body (experimental study)

Introduction. Damage to the liver under the conditions of long-term exposure to coal-rock dust on the body leads to significant impairment of its metabolic, detoxification, synthetic, and other functions. In this regard, it is relevant to search for early methods of prevention and correction of developing pathological changes in this organ. The purpose of the study is to experimentally study the effect of a complex drug with dihydroquercetin on the activity of free radical processes in the liver of laboratory rats over the dynamics of long-term exposure to coal-rock dust. Materials and methods. The work was carried out on male white rats weighing 200–250 g. The control group was kept under the standard vivarium conditions. The experimental group included rats that inhaled coal-rock dust in a priming chamber daily for 4 hours for 1, 3, 6, and 9 weeks and rats that received a daily complex drug with dihydroquercetin immediately before dust exposure. In the liver tissue, the activity of free radical oxidation and antioxidant defense enzymes (superoxide dismutase and catalase) was determined. Results. Priming rats with coal-rock dust for three weeks led to the gain in the intensity of free radical processes in the liver tissue: the initial level of accumulation of TBA-active oxidation products reliably raised by almost 2 times over the 1st week and 3 times over the 3rd week of the experiment compared to the control. An increase in the intensity of free radical oxidation was accompanied by activation of antioxidant defense enzymes - catalase - by 2.4 times at the 1st week and 1.6 times at the 3rd week of dust exposure. Raising the period of priming with dust up to 9 weeks led to multidirectional changes in oxidative metabolism in the liver tissue: at week 6, a decrease in the activity of superoxide dismutase was almost 3 times; at week 9, to a slight activation of antioxidant enzymes: superoxide dismutase by 55% and catalase by 36%.The most effective use of a complex drug with dihydroquercetin had been shown in short-term mode for three weeks, but not long-term (more than 6 weeks). The maximum antioxidant defense effect of dihydroquercetin was detected at the 3rd week of dust exposure – the level of free radical products decreased by 2 times and the activity of superoxide dismutase increased by 4 times. Limitations. Since the issue of organ-specific effects and the optimal duration of use of dihydroquercetin to increase resistance to free radical damage has been little elaborated, this imposes certain restrictions on its use for the correction and prevention of occupational diseases. Conclusion. The results obtained should be taken into account with long-term use of exogenous antioxidants in the correction and prevention of occupational diseases to support the level of endogenous antioxidant systems in the cell and the protective effect of dihydroquercetin for tissues most sensitive to the action of free radical processes.