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Cost and availability have often dictated the use of heterologous/alien prolactins in experiments, particularly in vivo. The assumption has been that what is initiated in the target cell is representative of the homologous hormone since many heterologous mammalian prolactins bind to and activate rodent receptors. Here, we examined gene expression in mouse liver in response to a 7‐day treatment with recombinant mouse prolactin (mRecPRL), recombinant ovine prolactin (oRecPRL) and pituitary extract ovine prolactin (oPitPRL). Having established mouse ribosomal protein S9 as the most stable reference gene in the liver in the absence and presence of prolactin treatment, we examined expression of the two most highly expressed prolactin receptors (PRLRs) and three members of the Cyp3a group of cytochrome P450 isoenzymes by qRTPCR. For short form (SF) 3 PRLR, mRecPRL doubled expression while for oRecPRL and oPitPRL expression was only 1.3‐fold control. For the long form (LF) PRLR, changes were similar to those seen for SF 3 PRLR, such that the SF3:LF PRLR ratio remained the same. Expression of the Cyp3as was also dependent on the prolactin origin and, although mRecPRL always stimulated, the other PRLs caused varying results. Compared to control, Cyp3a16 was stimulated 12‐fold by mRecPRL, 3‐fold by oRecPRL, and 6‐fold by oPitPRL. For Cyp3a41, mRecPRL was 3.7‐fold control, oRecPRL was without effect, and oPitPRL was 2‐fold control. Importantly, for Cyp3a44, mRecPRL stimulated 2‐fold, whereas both oRecPRL and oPitPRL had an opposite, inhibitory effect, with expression at 0.5‐fold control. We conclude that homologous hormone had the largest stimulatory effect on expression of all measured genes and that by contrast heterologous hormone showed reduced activity, no activity, or opposite activity, depending on the gene being analyzed. Thus, experimentation using alien heterologous PRL may lead to inaccurate conclusions. [ABSTRACT FROM AUTHOR]

Xylotrechus rusticus is one of the major forest trunk borers in China, causing a significant threat to afforestation and forestry engineering construction. Controlling X. rusticus is challenging because of its hidden life habit. The use of natural enemies against X. rusticus is an important strategy for controlling Cerambycidae pests. This is the first study to investigate the effectiveness of Dastarcus helophoroides as a biological control against X. rusticus under indoor and outdoor environments. The expression patterns of CYP6BK3 and CYP6BQ13 genes in various tissues and at different development stages were analyzed to clarify their role in the development of D. helophoroides. The results showed that the parasitism rate of D. helophoroides was the higher when their eggs were released on the larvae of X. rusticus L. indoors. Furthermore, the parasitism rate of D. helophoroides was the highest (86.67%) when their eggs were released on the larvae of X. rusticus in the ratio of 1:100 (host: D. helophoroides) under artificial wood segment simulation. The average parasitism rates of D. helophoroides eggs and adults on the exposed wood segment of the Cerambycidae were 81.83% and 83.29%, respectively. Indoor release of Scleroderma guani increased their parasitic rate on the aged larvae and pupae of X. rusticus. qPCR analysis of CYP450 gene expression showed that CYP6BK3 and CYP6BQ13 genes were expressed at all development stages of D. helophoroides; however, their expression levels were higher in adults than in the larvae. Notably, CYP6BK3 expression was the highest in the adipose tissue, whereas CYP6BQ13 was the highest in the hindgut tissue. [ABSTRACT FROM AUTHOR]

In toxicogenetics, an integrative approach including the prediction of phenotype based on post-mortem genotyping of drug-metabolising enzymes might help explain the cause of death (CoD) and manner of death (MoD). The use of concomitant drugs, however, might lead to phenoconversion, a mismatch between the phenotype based on the genotype and the metabolic profile actually observed after phenoconversion. The aim of our study was to evaluate the phenoconversion of CYP2D6, CYP2C9, CYP2C19, and CYP2B6 drug-metabolising enzymes in a series of autopsy cases tested positive for drugs that are substrates, inducers, or inhibitors of these enzymes. Our results showed a high rate of phenoconversion for all enzymes and a statistically significant higher frequency of poor and intermediate metabolisers for CYP2D6, CYP2C9, and CYP2C19 after phenoconversion. No association was found between phenotypes and CoD or MoD, suggesting that, although phenoconversion might be useful for a forensic toxicogenetics approach, more research is needed to overcome the challenges arising from the post-mortem setting. [ABSTRACT FROM AUTHOR]

In toxicogenetics, an integrative approach including the prediction of phenotype based on post-mortem genotyping of drug-metabolising enzymes might help explain the cause of death (CoD) and manner of death (MoD). The use of concomitant drugs, however, might lead to phenoconversion, a mismatch between the phenotype based on the genotype and the metabolic profile actually observed after phenoconversion. The aim of our study was to evaluate the phenoconversion of CYP2D6, CYP2C9, CYP2C19, and CYP2B6 drug-metabolising enzymes in a series of autopsy cases tested positive for drugs that are substrates, inducers, or inhibitors of these enzymes. Our results showed a high rate of phenoconversion for all enzymes and a statistically significant higher frequency of poor and intermediate metabolisers for CYP2D6, CYP2C9, and CYP2C19 after phenoconversion. No association was found between phenotypes and CoD or MoD, suggesting that, although phenoconversion might be useful for a forensic toxicogenetics approach, more research is needed to overcome the challenges arising from the post-mortem setting.

This study aimed to assess the associations between maternal drug use, cytochrome P450 (CYP450) genetic polymorphisms, and their interactions with the risk of congenital heart defects (CHDs) in offspring. A case-control study involving 569 mothers of CHD cases and 652 controls was conducted from November 2017 to January 2020. After adjusting for potential confounding factors, the results show that mothers who used ovulatory drugs (adjusted odds ratio [ aOR ] = 2.12; 95% confidence interval [ CI ]: 1.08–4.16), antidepressants (a OR = 2.56; 95% CI : 1.36–4.82), antiabortifacients (a OR = 1.55; 95% CI : 1.00–2.40), or traditional Chinese drugs (a OR = 1.97; 95% CI : 1.26–3.09) during pregnancy were at a significantly higher risk of CHDs in offspring. Maternal CYP450 genetic polymorphisms at rs1065852 (A/T vs. A/A: OR = 1.53, 95% CI : 1.10–2.14; T/T vs. A/A: OR = 1.57, 95% CI : 1.07–2.31) and rs16947 (G/G vs. C/C: OR = 3.41, 95% CI: 1.82–6.39) were also significantly associated with the risk of CHDs in offspring. Additionally, significant interactions were observed between the CYP450 genetic variants and drug use on the development of CHDs. In those of Chinese descent, ovulatory drugs, antidepressants, antiabortifacients, and traditional Chinese medicines may be associated with the risk of CHDs in offspring. Maternal CYP450 genes may regulate the effects of maternal drug exposure on fetal heart development. [ABSTRACT FROM AUTHOR]

Saffron is a well-known Chinese traditional herb, and crocin biosynthesis is related to the yield and quality of saffron. This study aimed to screen differentially expressed genes (DEGs) in saffron at different flowering stages and identify cytochrome P450 (CYP) genes involved in crocin biosynthesis. Saffron samples at different flowering stages were used for RNA sequencing, and DEGs between the samples at three days before the flowering stage (− 3da) and two days after the flowering stage (+ 2da) were screened. Thereafter, significantly differentially expressed CYP genes were identified, and CYP gene expression at different flowering stages and in various tissues of saffron was determined using real-time quantitative polymerase chain reaction (RT-qPCR). After sequencing and analysis, 1508 DEGs between the samples at − 3da and + 2da were identified, including 487 upregulated and 1021 downregulated genes, which were enriched in 16 biological processes, 5 cellular components, 3 molecular functions, and 11 KEGG pathways, including protein processing in endoplasmic reticulum, pentose and glucuronate interconversions, starch and sucrose metabolism, estrogen signaling pathway, and mitogen-activated protein kinase signaling pathway. In addition, 12 significantly differentially expressed CYP genes were identified. The RT-qPCR results showed that CYP76C4, CYP72A15, CYP72A219, CYP97B2, CYP714C2, CYP71A1, CYP94C1, and CYP86A8 were all expressed in the pistils, and CYP72A219, CYP72A15, CYP97B2, CYP71A1, and CYP86A8 were highly expressed in the pistils. Our study established a transcriptome library of saffron and found that CYP72A219, CYP72A15, CYP97B2, CYP71A1, and CYP86A8 may be candidates involved crocin biosynthesis in saffron. [ABSTRACT FROM AUTHOR]

The cytochrome P450 monooxygenases of insects play crucial roles in the metabolic detoxification of insecticides. Our previous finding showed that two cytochrome P450 genes, both CYP301B1 and CYP6AX1v2 , in the BPH underwent overexpression due to β-asarone. In this study, we investigated the molecular characteristics, expression patterns and functions of these two cytochrome P450 genes. The results showed that CYP301B1 had the highest expression level in the eggs, while CYP6AX1v2 was expressed in macropterous female adults. Moreover, the expression level of CYP301B1 in the head was higher than that in the integument, fat body and gut. The expression level of CYP6AX1v2 in the fat body and gut was higher than that in head and integument. Importantly, silencing CYP301B1 and CYP6AX1v2 separately could increase the sensitivity, resulting in significant higher mortality of BPH following treatment with β-asarone. Our findings indicated that CYP301B1 and CYP6AX1v2 could contribute to the resistance of BPH to β-asarone, and these two genes may be involved in the detoxification metabolism of β-asarone in BPH. • CYP301B1 is mainly expressed in the eggs, head and integument. • CYP6AX1v2 is mainly expressed in macropterous female adults and the fat body and gut. • Silencing CYP301B1 and CYP6AX1v2 could increase the sensitivity of BPH to β-asarone. [ABSTRACT FROM AUTHOR]

Aim: The association between cytochrome P450 (CYP) gene polymorphisms and anti-tuberculosis drug-induced hepatotoxicity (ATDH) was investigated in patients with or without pre-existing liver diseases (PLD). Materials & methods: We followed 164 tuberculosis subjects, 58 with PLD and 106 without PLD. Polymorphisms in CYP2D6, CYP2C9, CYP2C19, CYP3A4 and CYP3A5 were analyzed using the TaqMan ® SNP genotyping assay. Results: The CYP3A4*18 heterozygous genotype was associated with ATDH (OR: 3.24, 95% CI: 1.06-9.86) regardless of PLD presence. Among subjects without PLD, CYP3A4*18 heterozygotes had significantly higher ATDH risk (OR: 9.10, 95% CI: 1.56-53.16). Conversely, in the PLD group, CYP3A4*18 heterozygotes had lower ATDH risk (OR: 0.21, 95% CI: 0.05-0.98). Conclusion: CYP3A4*18 genotype is linked to ATDH in tuberculosis patients, with differential effects based on PLD presence.

Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), adults were collected from eight citrus groves across central Florida, and the level of insecticide resistance to ten insecticides was measured using a bottle bioassay. The gene expression of five cytochrome P450 CYP4 (CYP4C67, CYP4DA1, CYP4DB1, CYP4G70 and CYP4C68) and three glutathione S‐transferase (GSTD1, GSTE2 and GSTE1) genes was characterized in seven field populations of D. citri and compared with a laboratory population. Additionally, we reared four neonicotinoid insecticide resistant field populations in the laboratory and observed susceptibility changes without exposure to insecticides over multiple generations. The eight field populations of D. citri adults showed no and very low levels of resistance (RR = 1 and 2–10) to dimethoate, chlorprifos, carbaryl, fenpropathrin, bifenthrin, flupyradifurone and spinetoram. Very low to low resistance was found to imidacloprid and cyantraniliprole (RR = 2–10 and RR = 10–20). Moderate to high resistance was found for thiamethoxam (RR = 20–50 and RR = 50–100). The CYP4G70 and CYP4C68 genes were expressed at a higher level in field populations as compared with the laboratory population. Also the Davenport, Florida field population exhibited higher expression of all target genes compared to the laboratory population. Susceptibility to imidacloprid and thiamethoxam increased by 6.62‐ and 6.42‐fold, respectively, compared to the levels initially observed in the field over six generations of breeding without exposure. These results indicate that insecticide resistance may reverse in the field if insecticide selection pressure is removed from the spray schedule or with use of a rotational scheme with insecticides of different modes of action. Also, the results support use of insecticide resistance survey program combined with effective rotation for integrated insecticide resistance management of D. citri where huanglongbing (HLB) management includes vector suppression with insecticides. [ABSTRACT FROM AUTHOR]

Whole transcriptomes of the rotifer Brachionus plicatilis were analyzed using an Illumina sequencer. De novo assembly was performed with 49,122,780 raw reads using Trinity software. Among the assembled 42,820 contigs, 27,437 putative open reading frame contigs were identified (average length 1235 bp; N50 = 1707 bp). Functional gene annotation with Gene Ontology and InterProScan, in addition to Kyoto Encyclopedia of Genes and Genomes pathway analysis, highlighted the metabolism of xenobiotics by cytochrome P450 (CYP). In addition, 28 CYP genes were identified, and their transcriptional responses to benzo[ α ]pyrene (B[ α ]P) were investigated. Most of the CYPs were significantly upregulated or downregulated ( P < 0.05) in response to B[ α ]P, suggesting that Bp-CYP genes play a crucial role in detoxification mechanisms in response to xenobiotics. This study sheds light on the molecular defense mechanisms of the rotifer B. plicatilis in response to exposure to various chemicals. [ABSTRACT FROM AUTHOR]

To investigate the impact of cytochrome P450 (CYP) genetic polymorphisms CYP2B6, CYP2C19, and CYP3A5 on mRNA expression, cyclophosphamide/4-hydroxycyclophosphamide pharmacokinetics, and treatment outcomes of the R-CHOP regimen (rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone) in Chinese patients with non-Hodgkin's lymphoma, 567 cases were investigated. Plasma concentrations of cyclophosphamide/4-hydroxycyclophosphamide were determined using liquid chromatography-tandem mass spectrometry and pharmacokinetic parameters calculated. The frequencies of CYP2B6*1/*29 and CYP2B6*1/*30 were 18 (3.2%) and 30 (5.3%), respectively. Liver samples with CYP2B6*29/*30, CYP2C19*2, CYP2B6*6, or CYP3A5*3 had significantly lower target mRNA expression. Samples with CYP2B6*6 and/or CYP2C19*2 had lower exposure to 4-hydroxycyclophosphamide, whereas those with CYP2B6*1G/*1H had higher exposure. Multivariate model showed that samples with CYP2C19*2 or CYP2B6 785A>raG had worse treatment response than samples with CYP3A5*3. CYP2C19*2, CYP2B6*6, CYP2B6*29, and CYP2B6*30 were protective factors for toxicity in the multivariate model. The best model for gene-gene interactions for treatment response contained CYP2C19*2, CYP2B6 785A>G, and CYP3A5*3 (cross-validation consistency [CVC], 8/10; P = .0035). The best prediction model for grade 2-4 toxicities had CYP2C19*2, CYP2B6 785A>G, and 516 G>T (CVC, 10/10; P < .0001). In previous reports, we were the first to report that the frequency of copy-number variations in CYP2B6 is higher among Chinese than among other ethnic populations. Genetic variations in CYP2B6, CYP2C19, and CYP3A5 dramatically affected the mRNA expression of proteins, the pharmacokinetics of 4-hydroxycyclophosphamide, and the R-CHOP treatment response in Chinese subjects. Patients carrying CYP3A5*3 were more likely to have a better treatment response, whereas patients with CYP2C19*2 or CYP2B6 516G>T, or CYP2B6 785A>G had higher tolerance to treatment. [ABSTRACT FROM AUTHOR]

Pyrethroid resistance was one of the main reasons for control failure of cotton bollworm Helicoverpa armigera (Hübner) in China. The promotion of Bt crops decreased the application of chemical insecticides in controlling H. armigera . However, the cotton bollworm still kept high levels of resistance to fenvalerate. In this study, the resistance levels of 8 field-collected strains of H. armigera from north of China to 4 insecticides, as well as the expression levels of related P450 genes were investigated. The results of bioassay indicated that the resistance levels to fenvalerate in the field strains varied from 5.4- to 114.7-fold, while the resistance levels to lambda-cyhalothrin, phoxim and methomyl were low, which were ranged from 1.5- to 5.2-, 0.2- to 1.6-, and 2.9- to 8.3- fold, respectively, compared to a susceptible strain. Synergistic experiment showed that PBO was the most effective synergist in increasing the sensitivity of H. armigera to fenvalerate, suggesting that P450 enzymes were involved in the pyrethroid resistance in the field strains. The results of quantitative RT-PCR indicated that eight P450 genes ( CYP332A1 , CYP4L11 , CYP4L5 , CYP4M6 , CYP4M7 , CYP6B7 , CYP9A12 , CYP9A14 ) were all significantly overexpressed in Hejian1 and Xiajin1 strains of H. armigera collected in 2013, and CYP4L5 was significantly overexpressed in all the 6 field strains collected in 2014. CYP332A1 , CYP6B7 and CYP9A12 had very high overexpression levels in all the field strains, indicating their important roles in fenvalerate resistance. The results suggested that multiple P450 genes were involved in the high-level fenvalerate-resistance in different field strains of H. armigera collected from north of China. [ABSTRACT FROM AUTHOR]

The oriental armyworm Mythimna separata (Walker) is a devastating pest of cereal crops mainly in Asia and Oceania and recently become resistant to beta-cypermethrin (beta-CP). However, molecular biological studies of its response to beta-CP are scarce, and related genomic information is not available. In this study, we sequenced and de novo assembled the transcriptome of beta-CP susceptible M. separata (MsS-whole, abbr. MsS-W). A total of 30,486 unigenes were obtained, with an N50 length of 506 bp. A number of 21,051 unigenes were matched to public databases, of which 10,107 were classified into 59 gene ontology subcategories, 5792 were assigned into 25 clusters of orthologous groups of proteins subgroups and 12,123 were assigned to 257 Kyoto Encyclopedia of Genes and Genomes pathways. A total of 729 simple sequence repeats were detected. In addition, a total of 323 cytochrome P450-associated sequences from nine lepidopterous species, of which 130 were from M. separata, were analyzed using the maximum likelihood method and Bayesian inference. Among the 130 cytochrome P450-associated sequences from M. separata, 60 were dropped into CYP3 clan, which is associated with metabolizing xenobiotics and plant natural compounds. Furthermore, the beta-CP susceptible (MsS-2) and resistant (MsR-2) M. separata population transcriptomes were sequenced. Certain critical genes involved in beta-CP detoxification were detected and verified by quantitative real-time polymerase chain reaction. Collectively, our results provided a basis for further studies of the molecular mechanism of insecticide resistance in M. separata., (© 2022 Wiley Periodicals LLC.)

Background: Traumatic brain injury (TBI) induces arachidonic acid (ArA) release from cell membranes. ArA metabolites form a class of over 50 bioactive eicosanoids that can induce both adaptive and/or maladaptive brain responses. The dynamic metabolism of ArA to eicosanoids, and how they affect the injured brain, is poorly understood due to their diverse activities, trace levels, and short half-lives. The eicosanoids produced in the brain postinjury depend upon the enzymes present locally at any given time. Eicosanoids are synthesized by hemecontaining enzymes, including cyclooxygenases, lipoxygenases, and arachidonate monoxygenases. The latter comprise a subset of the cytochrome P450 "Cyp" gene family that metabolize fatty acids, steroids, as well as endogenous and exogenous toxicants. However, for many of these genes neither baseline neuroanatomical nor injury-related temporal expression have been studied in the brain. In a rat model of parietal cortex TBI, Cyp and eicosanoid-related mRNA levels were determined at 6 h, 24 h, 3d, and 7d postinjury in parietal cortex and hippocampus, where dynamic changes in eicosanoids have been observed. Quantitative real-time polymerase chain reaction with low density arrays were used to assay 62 rat Cyps, 37 of which metabolize ArA or other unsaturated fatty acids; 16 eicosanoid-related enzymes that metabolize ArA or its metabolites; 8 eicosanoid receptors; 5 other inflammatory- and recovery-related genes, plus 2 mouse Cyps as negative controls and 3 highly expressed "housekeeping" genes. Results: Sixteen arachidonate monoxygenases, 17 eicosanoid-related genes, and 12 other Cyps were regulated in the brain postinjury (p < 0.05, Tukey HSD). Discrete tissue levels and distinct postinjury temporal patterns of gene expression were observed in hippocampus and parietal cortex. Conclusions: The results suggest complex regulation of ArA and other lipid metabolism after TBI. Due to the temporal nature of brain injury-induced Cyp gene induction, manipulation of each gene (or its products) at a given time after TBI will be required to assess their contributions to secondary injury and/or recovery. Moreover, a better understanding of brain region localization and cell type-specific expression may be necessary to deduce the role of these eicosanoid-related genes in the healthy and injured brain. [ABSTRACT FROM AUTHOR]

The differential transcript patterns of five antioxidant genes, four genes related to the ginsenoside pathway and five P450 genes related to defense mechanism were investigated in in vitro adventitious roots of Panax ginseng after exposure to two different concentrations of heavy metals for 7 days. PgSOD- 1 and PgCAT transcription increased in a dose-dependent manner during the exposure to CuCl, NiCl, and CdCl, while all other tested scavenging enzymes didn't show significant increase during heavy metal exposure. Conversely, the mRNA transcripts of PgSQE, PgDDS were highly responsive to CuCl compared to NiCl exposure. However, the transcript profile of Pgβ- AS was highly induced upon NiCl treatment compared to CuCl and CdCl exposure. The expressions of PgCYP716A42, PgCYP71A50U, and PgCYP82C22 were regulated in similar manners, and all showed the highest transcript profile at 100 μM of CuCl, CdCl, and NiCl except PgCYP71D184, which showed the highest transcript level when subjected to 10 μM CuCl and NiCl. Thus it may suggest that in P. ginseng heavy metal interaction on cell membrane induced expression of various defense related genes via jasmonic acid pathway and also possesses cross talk networks with other defense related pathways. [ABSTRACT FROM AUTHOR]

Abstract: Cytochrome P450 (CYP) is a multifunctional super gene family, some of which play an important role in pesticide resistance. In this research, we used a real-time quantitative RT-PCR method, and a novel strategy, to measure the transcriptional level per gene copy using an exogenous RNA reference and DNA reference. The transcription levels of eight P450 genes in different tissues of fifth instar Bombyx mori larvae and their responses to pesticides (dichlorvos and deltamethrin) and sodium fluoride (NaF) were investigated. The results show different levels and patterns of expression of the different P450s in the various tissues observed. The P450s can be induced by pesticide and fluoride, but their responses to each are different. The results of this research are helpful in studying the tissue-specific expression of P450s in B. mori, and in developing new pesticide resistant silkworm varieties. [Copyright &y& Elsevier]

Abstract: Phenolic compounds are ubiquitous in plants which collectively synthesize several thousand different chemical structures characterized by hydroxylated aromatic ring(s). These compounds play several important functions in plants. They represent a striking example of metabolic plasticity enabling plants to adapt to changing biotic and abiotic environments and provide to plant products colour, taste, technological properties and putative health promoting benefits. Phenolic compounds represent the most studied phytochemicals and have been widely exploited as model systems in different areas of plant research. Initial studies in the field concerned the analytical characterization of a wide range of structures and of relevant enzymes with PAL being one of the most studied plant enzymes. This research is still active due to the complexity of the structures and the biosynthetic pathways As an example, the nature and functions of enzymes involved in lignin synthesis have been revisited several times, even in recent years. More recently, molecular biology and genomics have provided additional understanding of the mechanisms underlying the synthesis of these compounds with special emphasis on the regulation of gene expression by environmental factors. The extensive characterization of genes encoding the different enzymatic steps of flavonoid synthesis and cytochrome P450 genes have been among the most recent advances in this area. Metabolic engineering of lignins and flavonoids has been deeply investigated. Significant positive results have been obtained in both areas but the negative European opinion towards genetically modified organisms has considerably hampered potential applications. From a more basic point of view, global approaches (such as transcript and metabolite profiling) have investigated the repercussions of these engineered modulations of specific phenolics synthesis on other branches of plant metabolism. These studies have revealed a substantial and sometimes unexpected network of regulatory interactions. In the present time, the societal demand and an increasing interest for practical applications has stimulated a wide range of biological and epidemiological studies aiming at characterizing the health promoting properties of specific phenolic compounds with antioxidant activities towards cancer, cardiovascular and neurodegenerative diseases or for use in antiaging or cosmetic products. Increased emphasis on sustainable development should stimulate innovative investigations on phenolic synthesis for improving plant biomass and for a better control of plant and animal health. [Copyright &y& Elsevier]

Abstract In order to examine the usefulness of detoxifying genes as molecular markers in different chemical environments, isolation of cytochrome P450 genes (CYPs) belonging to the CYP4 family was performed in different samples from two subalpine populations of Daphnia pulex . The use of degenerate primers allowed us to isolate seven cDNAs. Four of them were assigned to the CYP4C subfamily, and were closely related to previously isolated crustacean CYP4s while the others were assigned to new CYP4AN and CYP4AP subfamilies. Expression studies, using semiquantitative polymerase chain reaction (PCR) followed by Southern blot hybridization with specific probes revealed differences in CYP4C32 and CYP4AP1 expressions between the two populations, which differ in the polyphenol richness of the vegetation surrounding their aquatic habitat. Further exposure to toxic dietary polyphenols showed different CYP induction patterns. Taken together, these preliminary results suggest a possible involvement of CYP4s in the ecological differentiation of subalpine D. pulex populations related to the polyphenol richness of the environmental vegetation. CYP4s may thus be considered as possible molecular markers in aquatic environmental bioreporting. [ABSTRACT FROM AUTHOR]

The primary structure of the cDNA clone SF28 was determined in sunflower ( Helianthus annuusL.) flowers. The clone comprises a 874-bp insert corresponding to 227 amino acid residues of the C-terminal part of the cytochrome P450 gene. The sunflower cytochrome P450 was considerably different from the already known plant and animal cytochromes P450. [ABSTRACT FROM AUTHOR]

Simple Summary: Childhood cancer patients receiving treatment containing alkylating agents are at risk of infertility, yet inter-individual variability in treatment-related ovarian damage is observed. Alkylating agents are metabolized by cytochrome P450 (CYP450) enzymes and polymorphisms in these CYP450 enzymes may explain this variability in ovarian damage. This study on genetic variation in CYP450 enzymes of chemotherapy-induced gonadotoxicity, using anti-Müllerian hormone (AMH) levels as a proxy for ovarian reserve, in female childhood cancer survivors (CCSs) may identify patients at risk of infertility. This unique global collaboration of two large CCS studies shows the significant gonadotoxic effect of enzyme CYP3A4*3 and significant protective effect of CYP2B6*2 on gonadal function in CCSs receiving alkylating agents. Genetic variation in CYP3A4 and CYP2B6 have previously been associated with gonadotoxicity after cancer treatment. These findings could guide risk prediction models determining patients at risk of chemotherapy-induced gonadal impairment. Background: Female childhood cancer survivors (CCSs) carry a risk of therapy-related gonadal dysfunction. Alkylating agents (AA) are well-established risk factors, yet inter-individual variability in ovarian function is observed. Polymorphisms in CYP450 enzymes may explain this variability in AA-induced ovarian damage. We aimed to evaluate associations between previously identified genetic polymorphisms in CYP450 enzymes and AA-related ovarian function among adult CCSs. Methods: Anti-Müllerian hormone (AMH) levels served as a proxy for ovarian function in a discovery cohort of adult female CCSs, from the pan-European PanCareLIFE cohort (n = 743; age (years): median 25.8, interquartile range (IQR) 22.1–30.6). Using two additive genetic models in linear and logistic regression, nine genetic variants in three CYP450 enzymes were analyzed in relation to cyclophosphamide equivalent dose (CED) score and their impact on AMH levels. The main model evaluated the effect of the variant on AMH and the interaction model evaluated the modifying effect of the variant on the impact of CED score on log-transformed AMH levels. Results were validated, and meta-analysis performed, using the USA-based St. Jude Lifetime Cohort (n = 391; age (years): median 31.3, IQR 26.6–37.4). Results: CYP3A4*3 was significantly associated with AMH levels in the discovery and replication cohort. Meta-analysis revealed a significant main deleterious effect (Beta (95% CI): −0.706 (−1.11–−0.298), p-value = 7 × 10−4) of CYP3A4*3 (rs4986910) on log-transformed AMH levels. CYP2B6*2 (rs8192709) showed a significant protective interaction effect (Beta (95% CI): 0.527 (0.126–0.928), p-value = 0.01) on log-transformed AMH levels in CCSs receiving more than 8000 mg/m2 CED. Conclusions: Female CCSs CYP3A4*3 carriers had significantly lower AMH levels, and CYP2B6*2 may have a protective effect on AMH levels. Identification of risk-contributing variants may improve individualized counselling regarding the treatment-related risk of infertility and fertility preservation options. [ABSTRACT FROM AUTHOR]

Nilaparvata lugens (Stål) is a serious pest of rice and has evolved different levels of resistance against most chemical pesticides. β-asarone is the main bioactive insecticidal compound of Acorus calamus L. that shows strong insecticidal activity against pests. In this study, we conducted a bioassay experiment to determine the contact toxicity of β-asarone to N. lugens nymphs. The LD 30 sublethal dose was 0.106 μg per nymph, with 95% confidence limits of 0.070–0.140 μg. We applied the LD 30 concentration of β-asarone to nymphs for 24 h or 72 h and then performed a transcriptome sequence analysis by referencing the N. lugens genome to characterize the variation. The transcriptomic analysis showed that several GO terms and KEGG pathways presented significant changes. Individually, 126 differentially expressed genes (DEGs), including 72 upregulated and 54 downregulated genes, were identified at 24 h, and 1771 DEGs, including 882 upregulated and 889 downregulated genes, were identified at 72 h. From the DEGs, we identified a total of 40 detoxification-related genes, including eighteen Cytochrome P450 monooxygenase genes (P450s), three Glutathione S-transferase genes, one Carboxylesterase gene, twelve UDP-glucosyltransferases and six ATP-binding cassette genes. We selected the eighteen P450s for subsequent verification by quantitative PCR. These findings indicated that β-asarone presented strong contact toxicity to N. lugens nymphs and induced obvious variation of detoxification-related genes that may be involved in the response to β-asarone. • β-asarone showed insecticidal activity against the brown planthopper (BPH) rice pest. • Application of β-asarone to BPH led to changes in the expression of 1897 genes. • Eighteen P450 genes may be involved in the response of BPH to β-asarone. [ABSTRACT FROM AUTHOR]

We identified 672, 374, and 379 CYPs proteins encoded by the CYPs genes in Gossypium hirsutum, Gossypium raimondii, and Gossypium arboreum, respectively. The genes were found to be distributed in all 26 chromosomes of the tetraploid cotton, with chrA05, chrA12, and their homeolog chromosomes harboring the highest number of genes. The physiochemical properties of the proteins encoded by the CYP450 genes varied in terms of their protein lengths, molecular weight, isoelectric points (pI), and even grand hydropathy values (GRAVY). However, over 99% of the cotton proteins had GRAVY values below 0, which indicated that the majority of the proteins encoded by the CYP450 genes were hydrophilic in nature, a common property of proteins encoded by stress-responsive genes. Moreover, through the RNA interference (RNAi) technique, the expression levels of Gh_D07G1197 and Gh_A13G2057 were suppressed, and the silenced plants showed a higher concentration of hydrogen peroxide (H2O2) with a significant reduction in the concentration levels of glutathione (GSH), ascorbate peroxidase (APX), and proline compared to the wild types under drought and salt stress conditions. Furthermore, the stress-responsive genes 1-Pyrroline–5-Carboxylate Synthetase (GhP5CS), superoxide dismutase (GhSOD), and myeloblastosis (GhMYB) were downregulated in VIGS plants, but showed upregulation in the leaf tissues of the wild types under drought and salt stress conditions. In addition, CYP450-silenced cotton plants exhibited a high level of oxidative injury due to high levels of oxidant enzymes, in addition to negative effects on CMS, ELWL, RLWC, and chlorophyll content The results provide the basic foundation for future exploration of the proteins encoded by the CYP450 genes in order to understand the physiological and biochemical mechanisms in enhancing drought and salt stress tolerance in plants. [ABSTRACT FROM AUTHOR]

The presence of polymorphisms in the CYP2D6 gene may modulate enzyme level and activity, thereby affecting individual responses to pharmacological treatment. Here, we compared the prevalence of the CYP2D6*10, *4, and 14* alleles in an Iranian population of different ethnicities with those of other populations. Allele and genotype frequency distributions of CYP2D6*10 variants and predicted phenotypes including extensive metabolizers, intermediate metabolizers, and poor metabolizers were analysed in blood samples of 300 unrelated healthy individuals in an Iranian population using polymerase chain reaction (PCR)-restriction fragment length polymorphism, PCR-single-strand conformation polymorphism, and direct genomic DNA sequencing. The CYP2D6*4 (G1846A) and *14 (G1758A) allelic frequencies were not detected in different ethnicities, demonstrating the absence of a significant contribution of these alleles in Iranian populations. However, the T/T, C/T, and C/C genotype frequencies of the CYP2D6*10 allele were significantly different (P<0.01) in all Iranian ethnic groups. Additionally, the frequency of the homozygous T/T variant of the CYP2D6*10 allele was significantly high in the Lure (P<0.017) and low in the Kurd (P<0.002) ethnicities. The frequency of the T/T variant of the CYP2D6*10 allele in central Iran was the highest (P<0.001), while the south of Iran had the lowest frequency (P<0.001). The frequency of the C/T variant of the CYP2D6*10 allele was significantly a bit high (P<0.001) in females compare to males, while the frequencies of the T/T variant in females is similar to males, which are 24.4% and 24.3%, respectively. In contrast to absence of the CYP2D6*4 (G1846A) and *14 (G1758A) alleles in Iranian populations of different ethnicities, the prediction of the CYP2D6*10 allele is required in drug research and routine treatment, where the information would be helpful for clinicians to optimize therapy or identify persons at risk of adverse drug reactions before clinical trials. Approximately 39.3% of subjects (24.3% homozygous T/T CYP2D6*10 as poor metabolizers and 15% heterozygous C/T CYP2D6*10 as intermediate metabolizers) had this allele; therefore, the harmful effects of drugs are relatively common among Iranians.