CYP1 Family Research Articles

Sequence diversity in the monooxygenases involved in oxime production in plant defense and signaling: a conservative revision in the nomenclature of the highly complex CYP79 family.

Cytochrome P450 monooxygenases of the CYP79 family catalyze conversion of specific amino acids into oximes feeding into a variety of metabolic plant pathways. Here we present an extensive phylogenetic tree of the CYP79 family built on carefully curated sequences collected across the entire plant kingdom. Based on a monophyletic origin of the P450s, a set of evolutionarily distinct branches was identified. Founded on the functionally characterized CYP79 sequences, sequence features of the individual substrate recognition sites (SRSs) were analyzed. Co-evolving amino acid residues were identified using co-evolutionary sequence analysis. SRS4 possesses a specific sequence pattern when tyrosine is a substrate. Except for the CYP79Cs and CYP79Fs, substrate preferences toward specific amino acids could not be assigned to specific subfamilies. The highly diversified CYP79 tree, reflecting recurrent independent evolution of CYP79s, may relate to the different roles of oximes in different plant species. The sequence differences across individual CYP79 subfamilies may facilitate the in vivo orchestration of channeled metabolic pathways based on altered surface charge domains of the CYP79 protein. Alternatively, they may serve to optimize dynamic interactions with oxime metabolizing enzymes to enable optimal ecological interactions. The outlined detailed curation of the CYP79 sequences used for building the phylogenetic tree made it appropriate to make a conservative phylogenetic tree-based revision of the naming of the sequences within this highly complex cytochrome P450 family. The same approach may be used in other complex P450 subfamilies. The detailed phylogeny of the CYP79 family will enable further exploration of the evolution of function in these enzymes.

The Synergistic and Opposing Roles of ω-Fatty Acid Hydroxylase (CYP4A11) and ω-1 Fatty Acid Hydroxylase (CYP2E1) in Chronic Liver Disease

Cytochrome P450 fatty acid hydroxylase consists of members of the CYP4 family that ω-hydroxylate fatty acids and the CYP2E1 that ω-1 hydroxylates fatty acids. Although ω and ω-1 hydroxylation of fatty acids have been thought to play a minor role in fatty acid metabolism (less than 20%), it plays a vital role in excess liver fatty acids overload seen in fasting, diabetes, metabolic disorder, and over-consumption of alcohol and high-fat diet. This pathway provides anabolic metabolites for gluconeogenesis, succinate, and acetate for lipogenesis. The CYP4A and CYP2E1 genes are activated in fasting and several metabolic disorders, suggesting a synergistic role in preventing fatty acid-induced lipotoxicity with the consequence of increased liver cholesterol and lipogenesis leading to increased Lipid Droplet (LD) deposition. During the progression of Metabolic Dysfunction-associated Steatotic Liver Disease (MASLD), activation of Phospholipase A2 (PLA2) releases arachidonic acid that CYP4A11 and CYP2E1 P450s metabolize to produce 20-hydroxyeicosatetraenoic acid (20-HETE) and 19-HETE, respectively. These metabolites have opposing roles in the progression of MASLD and chronic liver disease (CLD). This report discusses the synergistic role of the CYP4A and CYP2E1 P450s in the metabolism of saturated and unsaturated fatty acids and their opposite physiological role in the metabolism of Arachidonic Acid (AA). We finally discuss the role of ethanol in disrupting the synergistic and opposing roles of the CYP4A and CYP2E1 genes in MASLD and CLD.

Drug Metabolizing Enzymes: An Exclusive Guide into Latest Research in Pharmaco-genetic Dynamics in Arab Countries.

Drug metabolizing enzymes play a crucial role in the pharmacokinetics and pharmacodynamics of therapeutic drugs, influencing their efficacy and safety. This review explores the impact of genetic polymorphisms in drug-metabolizing genes on drug response within Arab populations. We examine the genetic diversity specific to Arab countries, focusing on the variations in key drug-metabolizing enzymes such as CYP450, GST, and UGT families. The review highlights recent research on polymorphisms in these genes and their implications for drug metabolism, including variations in allele frequencies and their effects on therapeutic outcomes. Additionally, the paper discusses how these genetic variations contribute to the variability in drug response and adverse drug reactions among individuals in Arab populations. By synthesizing current findings, this review aims to provide a comprehensive understanding of the pharmacogenetic landscape in Arab countries and offer insights into personalized medicine approaches tailored to genetic profiles. The findings underscore the importance of incorporating pharmacogenetic data into clinical practice to enhance drug efficacy and minimize adverse effects, ultimately paving the way for more effective and individualized treatment strategies in the region.

Biochemical and molecular characterization of pyrifluquinazon resistance in Bemisia tabaci Asia I

One of the most devastating polyphagous pests in the world, Bemisia tabaci (Gennadius), causes significant damage to a variety of crops and has demonstrated the ability to develop resistance to different classes of insecticides rapidly. A novel pyridine azomethine derivative pyrifluquinazon exhibits exceptional insecticidal toxicity against B. tabaci by interrupting the function of chordotonal receptor neurons. Formulating resistance management strategies is crucial to ensure the long-term use of this insecticide for whitefly control; however, the characteristics and possible mechanisms of pyrifluquinazon resistance in B. tabaci remain unclear. By employing pyrifluquinazon selection for 22 successive generations, the pyrifluquinazon-resistant strain (PQZ-R) was generated from a laboratory-susceptible population of B. tabaci Asia I (Lab-WB) and exhibited 39.65-fold resistance to pyrifluquinazon. When the realized heritability (h2) of B. tabaci to pyrifluquinazon was assumed to be the laboratory-estimated value (h2 = 0.181) and the mortality was 70–90%, only 12.3–22.6 generations were expected to be required to obtain a 10-fold increase in pyrifluquinazon resistance. While there was no significant cross-resistance to cyantraniliprole, dinotefuran, flonicamid, flupyradifurone, pymetrozine, sulfoxaflor, or thiamethoxam, the PQZ-R strain displayed slight cross-resistance to afidopyropen (3.14-fold). Synergism tests indicated that piperonyl butoxide (PBO) inhibits (4.36-fold) pyrifluquinazon resistance in the PQZ-R strain. This impact may be attributed to enhanced detoxification (elevated 3.91-fold) mediated by cytochrome P450 monooxygenase (P450). Compared to Lab-WB, the PQZ-R strain exhibited no significant overexpression of the 13 published detoxification-related P450 genes from CYP303, CYP4, and CYP6 families in B. tabaci. The combined knowledge gained from this study will enable further investigations into the function of qualitative and quantitative variations in P450-encoded genes, leading to innovative approaches for efficiently managing B. tabaci.

Identification and expression dynamics of CYPome across different developmental stages of Maconellicoccus hirsutus (Green)

Maconellicoccus hirsutus is a highly polyphagous insect pest, posing a substantial threat to various crop sp., especially in the tropical and sub-tropical regions of the world. While extensive physiological and biological studies have been conducted on this pest, the lack of genetic information has hindered our understanding of the molecular mechanisms underlying its growth, development, and xenobiotic metabolism. The Cytochrome P450 gene, a member of the CYP gene superfamily ubiquitous in living organisms is associated with growth, development, and the metabolism of both endogenous and exogenous substances, contributing to the insect's adaptability in diverse environments. To elucidate the specific role of the CYP450 gene family in M. hirsutus which has remained largely unexplored, a de novo transcriptome assembly of the pink mealybug was constructed. A total of 120 proteins were annotated as CYP450 genes through homology search of the predicted protein sequences across different databases. Phylogenetic studies resulted in categorizing 120 CYP450 genes into four CYP clans. A total of 22 CYP450 families and 30 subfamilies were categorized, with CYP6 forming the dominant family. The study also revealed five genes (Halloween genes) associated with the insect hormone biosynthesis pathway. Further, the expression of ten selected CYP450 genes was studied using qRT-PCR across crawler, nymph, and adult stages, and identified genes that were expressed at specific stages of the insects. Thus, the findings of this study reveal the expression dynamics and possible function of the CYP450 gene family in the growth, development, and adaptive strategies of M. hirsutus which can be further functionally validated.

Decoding the Role of CYP450 Enzymes in Metabolism and Disease: A Comprehensive Review.

Cytochrome P450 (CYP450) is a group of enzymes that play an essential role in Phase I metabolism, with 57 functional genes classified into 18 families in the human genome, of which the CYP1, CYP2, and CYP3 families are prominent. Beyond drug metabolism, CYP enzymes metabolize endogenous compounds such as lipids, proteins, and hormones to maintain physiological homeostasis. Thus, dysregulation of CYP450 enzymes can lead to different endocrine disorders. Moreover, CYP450 enzymes significantly contribute to fatty acid metabolism, cholesterol synthesis, and bile acid biosynthesis, impacting cellular physiology and disease pathogenesis. Their diverse functions emphasize their therapeutic potential in managing hypercholesterolemia and neurodegenerative diseases. Additionally, CYP450 enzymes are implicated in the onset and development of illnesses such as cancer, influencing chemotherapy outcomes. Assessment of CYP450 enzyme expression and activity aids in evaluating liver health state and differentiating between liver diseases, guiding therapeutic decisions, and optimizing drug efficacy. Understanding the roles of CYP450 enzymes and the clinical effect of their genetic polymorphisms is crucial for developing personalized therapeutic strategies and enhancing drug responses in diverse patient populations.

Changes of Active Substances in Ganoderma lucidum during Different Growth Periods and Analysis of Their Molecular Mechanism.

Ganoderma lucidum, renowned as an essential edible and medicinal mushroom in China, remains shrouded in limited understanding concerning the intrinsic mechanisms governing the accumulation of active components and potential protein expression across its diverse developmental stages. Accordingly, this study employed a meticulous integration of metabolomics and proteomics techniques to scrutinize the dynamic alterations in metabolite accumulation and protein expression in G. lucidum throughout its growth phases. The metabolomics analysis unveiled elevated levels of triterpenoids, steroids, and polyphenolic compounds during the budding stage (BS) of mushroom growth, with prominent compounds including Diplazium and Ganoderenic acids E, H, and I, alongside key steroids such as cholesterol and 4,4-dimethyl-5alpha-cholesta-8,14,24-trien-3beta-ol. Additionally, nutrients such as polysaccharides, flavonoids, and purines exhibited heightened presence during the maturation stage (FS) of ascospores. Proteomic scrutiny demonstrated the modulation of triterpenoid synthesis by the CYP450, HMGR, HMGS, and ERG protein families, all exhibiting a decline as G. lucidum progressed, except for the ARE family, which displayed an upward trajectory. Therefore, BS is recommended as the best harvesting period for G. lucidum. This investigation contributes novel insights into the holistic exploitation of G. lucidum.

Effect of High Temperature on Abamectin and Thiamethoxam Tolerance in Bemisia tabaci MEAM1 (Hemiptera: Aleyrodidae).

Bemisia tabaci (Gennadius) is one of the most important invasive species in China, with strong insecticide resistance and thermotolerance. In this study, we investigated the effects of elevated temperature on the tolerance of B. tabaci MEMA1 to abamectin (AB) and thianethixam (TH) insecticides. We firstly cloned two new CYP450 genes from B. tabaci MEAM1, including one CYP6 family gene (BtCYP6k1) and one CYP305 family gene (BtCYP305a1). The expression patterns of the two BtCYP450 genes were compared in response to high-temperature stress and insecticide exposure, and RNAi was then used to demonstrate the role that these two genes play in insecticide tolerance. The results showed that expression of the two BtCYP450 genes could be induced by exposure to elevated temperature or insecticide, but this gene expression could be inhibited to a certain extent when insects were exposed to the combined effects of high temperature and insecticide treatment. For AB treatment, the expression of the two BtCYP450 genes reached the lowest level when insects were exposed to a temperature of 41 °C and treated with AB (combined effects of temperature and insecticide). In contrast, TH treatment showed a general decrease in the expression of the two BtCYP450 genes with exposure to elevated temperatures. These findings suggest that insecticide tolerance in B. tabaci MEAM1 could be mediated by high temperatures. This study provides a prospective method for the more effective application of insecticides for the control of B. tabaci in the field.

Genome-Wide Identification, Characterization, and Expression Analysis of the CYP450 Family Associated with Triterpenoid Saponin in Soapberry (Sapindus mukorossi Gaertn.)

Soapberry (Sapindus mukorossi Gaertn.) is a tree species of the family Sapindaceae, the pericarp of which is rich in triterpenoid saponins, which are important in chemical production, biomedicine, and other fields. Cytochrome P450 monooxygenase (CYP450) is involved in the modification of the skeletons of triterpenoid saponins and is linked to their diversity. We previously identified 323 CYP450 genes in the transcriptome of soapberry and screened 40 CYP450 genes related to the synthesis of triterpenoid saponins by gene annotation and conserved structural domain analysis. The genetic structure and phylogeny of the CYP450 genes were analyzed separately. Phylogenetic analysis categorized the CYP450 genes of soapberry into five subfamilies, the members of which had similar conserved cumulative sequences and intron structures. A cis-acting element analysis implicated several genes in the responses to environmental changes and hormones. The expression of several genes during eight periods of fruit development was analyzed by real-time quantitative qRT-PCR; most showed high expression during the first four periods of fruit development, and their expression decreased as the fruits matured. A co-expression network analysis of SmCYP450s and related genes in the triterpenoid saponin synthesis pathway was performed. Correlation analysis showed that 40 SmCYP450s may be involved in saponin synthesis in soapberry. The triterpenoid saponin synthesis-related candidate genes identified in this study provide insight into the synthesis and regulation of triterpenoid saponins in soapberry.

Comparison of the Effect of Pruning on Plant Growth and Transcriptome Profiles in Different Tea Varieties

Although pruning contributes to the growth and development of new shoots, it is important to note that the growth potential and yield of tea varieties may differ after pruning due to genetic and environmental factors. In this experiment, 20 different varieties of tea plants were used to observe their potential for growth, shoot development, and other phenotypic indexes after pruning. The study aimed to determine the suitability of each variety for heavy pruning. It was concluded that there were obvious differences in tree strength and new growth after pruning of the different varieties, with ‘Zhongcha 302’ exhibiting the strongest growth and ‘Emei Wenchun’ showing the weakest growth. In order to understand the molecular mechanisms involved, a transcriptomic analysis was carried out on the two tea varieties. The results of the data indicate that the expression of CYP450 family was high in ‘Zhongcha 302’. In ‘Emei Wenchun’, the expression of NCED was higher than that in ‘Zhongcha 302’. The KEGG results indicate that pruning stimulates the expression of genes involved in phytohormone signalling and plant–pathogen interaction pathways in tea plants. The study offered scientific guidance for tea plant pruning suitability and preliminarily revealed the regulatory mechanism of new shoot growth in different tea plant varieties at the transcriptome level.

Brassinosteroids biosynthetic gene MdBR6OX2 regulates salt stress tolerance in both apple and Arabidopsis

Salt stress is a critical limiting factor for fruit yield and quality of apples. Brassinosteroids (BRs) play an important role in response to abiotic stresses. In the present study, application of 2,4- Epicastasterone on seedlings of Malus ‘M9T337’ and Malus domestica ‘Gala3’ alleviated the physiological effects, such as growth inhibition and leaf yellowing, induced by salt stress. Further analysis revealed that treatment with NaCl induced expression of genes involved in BR biosynthesis in ‘M9T337’ and ‘Gala3’. Among which, the expression of BR biosynthetic gene MdBR6OX2 showed a three-fold upregulation upon salt treatment, suggesting its potential role in response to salt stress in apple. MdBR6OX2, belonging to the CYP450 family, contains a signal peptide region and a P450 domain. Expression patterns analysis showed that the expression of MdBR6OX2 can be significantly induced by different abiotic stresses. Overexpressing MdBR6OX2 enhanced the tolerance of apple callis to salt stress, and the contents of endogenous BR-related compounds, such as Typhastero (TY), Castasterone (CS) and Brassinolide (BL) were significantly increased in transgenic calli compared with that of wild-type. Extopic expression of MdBR6OX2 enhanced tolerance to salt stress in Arabidopsis. Genes associated with salt stress were significantly up-regulated, and the contents of BR-related compounds were significantly elevated under salt stress. Our data revealed that BR-biosynthetic gene MdBR6OX2 positively regulates salt stress tolerance in both apple calli and Arabidopsis.

Identification of the cytochrome P450 gene AccCYP6A13 in Apis cerana cerana and its response to environmental stress

Cytochrome P450 plays a crucial role in regulating insect growth, development, and resisting a variety of stresses. Insect metamorphosis and response to external stress are altered by deleting CYP450 genes. In this study, we identified and analyzed a novel gene of CYP450 family, AccCYP6A13, from Apis cerana cerana, and explored its role in the response of Apis cerana cerana to adverse external stressors. It was found that the expression of AccCYP6A13 was spatiotemporal specificity. The expression level increased with age and reached its highest value in the adult stage. The primarily expressiong location were legs, brain, and epidermis of honeybees. Stress conditions can affect the expression of AccCYP6A13 depending on treatment times. RNA interference experiments have shown that knocking down AccCYP6A13 reduces antioxidant activity and deactivates detoxification enzymes, resulting in oxidative damage accumulation and a decline in detoxification capability in bees, as well as inhibiting numerous antioxidant genes. Additionally, knockdown of the AccCYP6A13 gene in Apis cerana cerana resulted in increased sensitivity to pesticides and increased mortality when treated with neonicotinoid pesticides such as thiamethoxam. AccCYP6A13 overexpression in a prokaryotic system further confirmed its role in resistance to oxidative stress. To summarize, AccCYP6A13 may play an essential role in the normal development and response to environmental stress in Apis cerana cerana. Furthermore, this study contributed to the theoretical understanding of bee resistance biology.

Abstract 7277: Discovery of INV-9956, an orally available and highly selective Cyp11A1 inhibitor for the treatment of castration-resistant prostate cancer with drug resistant AR mutations

Abstract Androgen Receptor (AR) signaling plays a profound role in prostate cancer and development. Efforts in harnessing androgen and AR axis for castration-resistant prostate cancer treatment have led to discovery of several therapeutics, including AR antagonists and Cyp17 inhibitors. However, resistance to these approved therapeutics has emerged, and remained a significant unmet medical need. Among the prominent resistance mechanisms of action are restored androgen signaling via AR amplification and overexpression, and gain-of-function mutations occurring at AR ligand binding domain that can be agonized by existing AR antagonists and/or non-androgenic steroids such as glucocorticoids. Targeting Cyp11A1 that catalyzes the formation of pregnenolone is effective to ablate biosynthesis of androgens and all steroids and thus holds promise to deliver coverage of a significant percentage of drug resistant AR mutations and benefit CRPC patients post treatment with AR antagonists or Cyp17 inhibitors. Here we present discovery and characterization of INV-9956, an orally available, selective, and potent Cyp11A1 inhibitor with well-acceptable pharmacokinetic properties. INV-9956 potently inhibits biosynthesis of pregnenolone and other androgens in H295R cells with IC50 at single-digit nanomolar, suppresses proliferation of prostate cancer cell lines harboring major clinically acquired drug resistant AR LBD mutations, such as AR L702H using coculture assays. Orally administered INV-9956 showed consistent dose dependent PK/PD correlations across animal species. Particularly, INV-9956 displayed pronounced tumor inhibition superior to known competitors in a CRPC VCaP xenograft model and in a “humanized” animal model. Furthermore, INV-9956 displayed a good tox profile in 14-day repeated dosing rat and dog studies, supported by a high selectivity to Cyp11A1 over other Cyp family members, clean hERG and mini-Ames, and no significant off-target activity in a safety panel screen. The promising preclinical profiles of INV-9956 lend strong support to its potentials in addressing unmet needs in CRPC treatments and warrant clinical development. Citation Format: Chang Bai, Danqi Chen, Zilei Xia, Shujuan Huang, LIshan Lin, Zhaopie Zeng, Jing Su, Yongkui Sun. Discovery of INV-9956, an orally available and highly selective Cyp11A1 inhibitor for the treatment of castration-resistant prostate cancer with drug resistant AR mutations [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7277.

Perfluorooctanoic acid inhibits cell proliferation through mitochondrial damage

Grown evidence has shown that the liver and reproductive organs were the main target organs of perfluorooctanoic acid (PFOA). Herein, we studied a toxic mechanism of PFOA using HeLa Chang liver epithelial cells. When incubated with PFOA for 24 h or 48 h, cell proliferation was inhibited in a concentration- and time-dependent fashion, but interestingly, the feature of dead cells was not notable. Mitochondrial volume was increased with concentration and time, whereas the mitochondrial membrane potential and produced ATP amounts were significantly reduced. Autophagosome-like vacuoles and contraction of the mitochondrial inner membrane were observed in PFOA-treated cells. The expression of acetyl CoA carboxylase (ACC) and p-ACC proteins rapidly decreased, and that of mitochondrial dynamics–related proteins increased. The expression of solute carrier family 7 genes, ChaC glutathione-specific gamma-glutamylcyclotransferase 1, and 5S ribosomal RNA gene was up-regulated the most in cells exposed to PFOA for 24 h, and the KEGG pathway analysis revealed that PFOA the most affected metabolic pathways and olfactory transduction. More importantly, PPAR alpha, fatty acid binding protein 1, and CYP450 family 1 subfamily A member 1 were identified as the target proteins for binding between PFOA and cells. Taken together, we suggest that disruption of mitochondrial integrity and function may contribute closely to PFOA-induced cell proliferation inhibition.

RNA Interference Reveals the Impacts of CYP6CY7 on Imidacloprid Resistance in Aphis glycines.

Cytochrome P450 (CYP) is a group of important detoxification enzymes found in insects related to their resistance to insecticides. To elucidate the CYP6 family genes of P450, which are potentially related to imidacloprid resistance in Aphis glycines, the CYP6 cDNA sequences of A. glycines were studied. The transcriptome of A. glycines was constructed, and the CYP6 cDNA sequences of A. glycines were screened. Their relative expression levels in response to imidacloprid induction were examined through qRT-PCR, and the CYP6s with higher expression levels were used to study the detoxification of imidacloprid through RNA interference and a bioassay. Twelve CYP6s were obtained from the A. glycines transcriptome. These samples were named by the International P450 Nomenclature Committee and registered in GenBank. After 3, 6, 12, 24 and 48 h of induction with LC50 concentrations of imidacloprid, the relative expression levels of these CYP6s increased; the expression level of CYP6CY7 experienced the highest increase, being more than 3-fold higher than that of those of the non-imidacloprid-induced CYP6s. After RNA interference for CYP6CY7, the relative expression level of CYP6CY7 significantly decreased after 3, 6 and 12 h, while the corresponding P450 enzyme activity decreased after 12 and 24 h. The mortality of A. glycines due to imidacloprid treatment increased by 14.71% at 24 h. CYP6CY7 might detoxify imidacloprid in A. glycines. This study provides a theoretical basis for the further study of the mechanism of action of CYP6s and potential new methods for improving insecticidal efficacy.

Effects of deltamethrin exposure on the cytochrome P450 monooxygenases of Aedes albopictus (Skuse) larvae from a dengue-endemic region of northern part of West Bengal, India.

Aedes albopictus is highly prevalent in the northern part of West Bengal and is considered to be responsible for the recent dengue outbreaks in this region. Control of this vector is largely relied on the use of synthetic pyrethroids, which can lead to the development of resistance. In the present study, larvae of three wild Ae. albopictus populations from the dengue-endemic regions were screened for deltamethrin resistance, and the role of cytochrome P450 monooxygenases (CYPs) was investigated in deltamethrin exposed and unexposed larvae. Two populations were incipient resistant, and one population was completely resistant against deltamethrin. Monooxygenase titration assay revealed the involvement of CYPs in deltamethrin resistance along with an induction effect of deltamethrin exposure. Gene expression studies revealed differential expression of five CYP6 family genes, CYP6A8, CYP6P12, CYP6A14, CYP6N3 and CYP6N6, with high constitutive expression of CYP6A8 and CYP6P12 in all the populations before and after deltamethrin exposure. From these findings, it was evident that CYPs play an important role in the development of deltamethrin resistance in the Ae. albopictus populations in this region.

Monitoring Aedes populations for arboviruses, Wolbachia, insecticide resistance and its mechanisms in various agroecosystems in Benin

Aedes mosquitoes are the main vectors of arboviruses in Benin. Cases of dengue have been reported in Benin with all four serotypes of the virus actively circulating in this region. Some agricultural settings are known to harbor Aedes vectors responsible for the transmission of arboviruses. The massive use of certain insecticides in agricultural settings has probably contributed to insecticide resistance in these vectors. In Benin, the susceptibility of arbovirus vectors to insecticides is poorly studied. In addition, the distribution of Wolbachia spp., which is used against some arboviruses is unknown. Moreover, there is limited information regarding the vectors responsible for the transmission of arboviruses in Benin. This present study monitored the species composition, arboviruses, and Wolbachia symbiont status, as well as the phenotypic and molecular insecticide resistance profile of Aedes populations from three agroecosystems in Benin. Aedes species identification was performed morphologically and confirmed using qPCR. (RT)-qPCR assay was applied for monitoring the presence of DENV, CHIKV, ZIKV, and WNV pathogens as well as for naturally occurring Wolbachia symbionts. Insecticide resistance was assessed phenotypically, by permethrin (0.75%) exposure of Adults (F0) using World Health Organization (WHO) bioassay protocols, and at the molecular level, using TaqMan (RT)-qPCR assays for assessing knock-down resistance (kdr) mutations (F1534C, V1016G/I, and S989P) and the expression levels of eight detoxification genes (P450s from the CYP9 and CYP6 families, carboxylesterases and glutathione-S-transferases). Aedes aegypti (Ae. aegypti) mosquitoes were the most abundant (93.9%) in the three agroecosystems studied, followed by Aedes albopictus (Ae. albopictus) mosquitoes (6.1%). No arboviruses were detected in the study's mosquito populations. Naturally occurring Wolbachia symbionts were present in 7 pools out of 15 pools tested. This could influence the effectiveness of vector control strategies based on exogenously introduced Wolbachia, all present in the three agroecosystems. Full susceptibility to permethrin was observed in all tested populations of Ae. albopictus. On the contrary, Ae. aegypti were found to be resistant in all three agroecosystem sites except for banana plantation sites, where full susceptibility was observed. Molecular analysis revealed that individual target site resistance kdr mutations F1534C and V1016G/I were detected in most Ae. aegypti populations. Additionally, double mutant (F1534C + V1016G/I) mosquitoes were found in some populations, and in one case, triple mutant (F1534C + V1016G/I + S989P) mosquitoes were detected. Metabolic resistance, as reflected by overexpression of three P450 genes (CYP6BB2, CYP9J26, and CYP9J32), was also detected in Ae. aegypti mosquitoes. Our study provides information that could be used to strategize future vector control strategies and highlights the importance of continuing vector surveillance. Future studies should assess the effect of piperonyl butoxide (PBO) on metabolic resistance and identify the different strains of Wolbachia spp., to choose the best vector control strategies in Benin.

Sequence similarity networks bear out hierarchical relationships of green cytochrome P450.

Land plants have diversified enzyme families. One of the most prominent is the cytochrome P450 (CYP or CYP450) family. With over 443,000 CYP proteins sequenced across the tree of life, CYPs are ubiquitous in archaea, bacteria, and eukaryotes. Here, we focused on land plants and algae to study the role of CYP diversification. CYPs, acting as monooxygenases, catalyze hydroxylation reactions crucial for specialized plant metabolic pathways, including detoxification and phytohormone production; the CYPome consists of one enormous superfamily that is divided into clans and families. Their evolutionary history speaks of high substrate promiscuity; radiation and functional diversification have yielded numerous CYP families. To understand the evolutionary relationships within the CYPs, we employed sequence similarity network analyses. We recovered distinct clusters representing different CYP families, reflecting their diversified sequences that we link to the prediction of functionalities. Hierarchical clustering and phylogenetic analysis further elucidated relationships between CYP clans, uncovering their shared deep evolutionary history. We explored the distribution and diversification of CYP subfamilies across plant and algal lineages, uncovering novel candidates and providing insights into the evolution of these enzyme families. This identified unexpected relationships between CYP families, such as the link between CYP82 and CYP74, shedding light on their roles in plant defense signaling pathways. Our approach provides a methodology that brings insights into the emergence of new functions within the CYP450 family, contributing to the evolutionary history of plants and algae. These insights can be further validated and implemented via experimental setups under various external conditions.

Genetic Polymorphisms in CYP2 Gene Family in Bulgarian Individuals and their Clinical Implications

Abstract The cytochrome P450 superfamily consists of hemeproteins involved in the detoxication of different xenobiotics, including drugs. The CYP2 gene family is responsible for the metabolism of 80% of the drugs in clinical use. There are considerable interindividual and interethnic variabilities in the rate of drug metabolism as a result of genetic polymorphisms. The goal of our study was to determine the frequency of 10 genetic polymorphisms in CYP2 family genes to give light on the pharmacogenetic defects of the main CYPs, involved in drug metabolism, in Bulgarian individuals. We detected high allele frequency for CYP2D6*10 (0.27), CYP2D6*4 (0.22), and CYP2B6*9 (0.24), followed by CYP2C19*2 (0.14), CYP2C9*3 (0.11) and CYP2C9*2 (0.09). The genotype frequencies were also determined for all investigated variants. In total 47.2% of the analyzed individuals carried CYP2D6 genetic polymorphisms – 5.6% carried a single variant and 41.6% were found to have two or more such variants. Homozygotes for CYP2D6 variants were established among 14% of Bulgarian individuals. Determination of the prevailing pharmacogenetic polymorphisms of the CYPs, most responsible for drug metabolism, will lead to a lower risk of drug toxicity, increased drug efficacy, and drug dose optimization.

Detrimental impacts of concomitant application of cadmium and pesticides are ameliorated by 24-epibrassinolide through alteration in oxidative status and CYP genes expression in Zea mays L.

The ameliorative impact of 24-epibrassinolide (EBL) was assessed under toxicity of cadmium (Cd) and three different pesticides including diazinon (DZ), imidacloprid (IMI), and chlorpyrifos (CPS) in maize plants. Maize seeds were pre-treated with 0.01 μM of EBL and grown in soil contaminated with 100 μM Cd for one month. Then, at five fully expanded leaf stage, plants received DZ, IMI, and CPS pesticides separately by foliar spray and were harvested five days later. The EBL seed primed plants, grown in soil contaminated with Cd and exposed to pesticides, exhibited higher deleterious photosynthesis (Pn) and stomatal conductivity (Gs) compared to un-EBL-treated plants. A considerable decrease was observed in the pesticide's residue and Cd, MDA and H2O2 contents of EBL + Cd + Pesticide treated plants compared to the Cd + Pesticide groups plants. Higher phenol, flavonoid and protein contents, as well as increment in SOD, CAT, POD, APX, PAL, GST, and GR enzymes activity were detected in EBL + Cd + Pesticide groups compared to the plants imposed with Cd + Pesticides. The expression analyses of four CYP family members including CYP71C1, CYP71C3v2, CYP72A5, and CYP81A9 revealed downregulation under Cd and Cd + pesticides treatments, while EBL seed priming highly induced transcript abundance of CYP71C1 (EBL + Cd + IMI); CYP71C3v2 (EBL + Cd + DZ & EBL + Cd + IMI) and CYP81A9 (EBL + Cd + DZ & EBL + Cd + CPS), specifically. The higher mRNA accumulation of CYP72A5 gene was exhibited under all three different EBL + Cd + pesticides treatments. The 24-epibrassinolide alleviates the adverse impacts of Cd and pesticides via the activation of enzymatic and non-enzymatic components to eliminate ROS through the up-regulation of CYP genes. This facilitates conjugation of glutathione to pesticides residue, leading to their rapid transportation into the vacuole or out of the membrane.