Insect chemical ecology (ICE) evolved as a discipline concerned with plant–insect interactions, and also with a strong focus on intraspecific pheromone-mediated communication. Progress in this field has rendered a more complete picture of how insects exploit chemical information in their surroundings in order to survive and navigate their world successfully. Simultaneously, this progress has prompted new research questions about the evolution of insect chemosensation and related ecological adaptations, molecular mechanisms that mediate commonly observed behaviors, and the consequences of chemically mediated interactions in different ecosystems. Themed meetings, workshops, and summer schools are ideal platforms for discussing scientific advancements as well as identifying gaps and challenges within the discipline. From the 11th to the 22nd of June 2018, the 11th annual PhD course in ICE was held at the Swedish University of Agricultural Sciences (SLU) Alnarp, Sweden. The course was made up of 35 student participants from 22 nationalities  as well as 32 lecturers. Lectures and laboratory demonstrations were supported by literature seminars, and four broad research areas were covered: (1) multitrophic interactions and plant defenses, (2) chemical communication focusing on odor sensing, processing, and behavior, (3) disease vectors, and (4) applied aspects of basic ICE research in agriculture. This particular article contains a summary and brief synthesis of these main emergent themes and discussions from the ICE 2018 course. In addition, we also provide suggestions on teaching the next generation of ICE scientists, especially during unprecedented global situations. Since the discovery of insect pheromones and the role of chemicals in plant–insect interactions, insect chemical ecology (ICE) has developed into a distinct research discipline being practiced at universities and other research organizations worldwide. Moreover, basic research in ICE has led to practical applications that contribute significantly to the control of agricultural or blood-feeding pests. High-quality education of PhD students in the field of ICE is of utmost importance for the discipline to grow and advance for future scientific discoveries as well as practical implementation. Recognizing the need for training and international networking of chemical ecologists, Bill Hansson and Ylva Hillbur initiated the first international ICE PhD course in 2005 at the Swedish University of Agriculture (SLU) involving teachers from Alnarp and Lund University, as well as many international lecturers. The interest of students and their positive response led to follow-up courses in 2007 and 2009 at Alnarp. In 2009, it was decided to rotate the location of the ICE courses and increase its frequency  to better accommodate the needs of international PhD students. As a result of this, a 2010 issue of the course was held at the Department of Entomology, Pennsylvania State University (PSU) and a subsequent one in 2012 at Max Planck Institute (MPI) for Chemical Ecology (CE) in Jena, Germany (where Bill Hansson had become the director and head of the Department of Neuroethology), merging resources and competence from three research institutions studying the chemical ecology of insects. With Teun Dekker at SLU , Tom Baker at PSU  and the Neuroethology group members  at MPI, the course continued being arranged in turns by the three host institutions strongly promoting teacher and student exchange across borders. After the 2018 course, which is the subject of the current report, the International Centre of Insect Physiology and Ecology (ICIPE) in Nairobi, Kenya, joined the consortium of organizing institutions and arranged the first ICE PhD course in Africa in 2019, illustrating the establishment and development of the course as an internationally recognized platform for education and exchange of students, junior and senior chemical ecologists.Plants, insects, and microbes are part of terrestrial ecosystems. Trophic interactions, such as the consumption of plant material by herbivorous insects, are key factors that lead to physiological and morphological adaptations in coexisting organisms. For example, plants have evolved multiple defense mechanisms against herbivory, and insects likewise evolve countermeasures against plant defenses  damaged by larvae of the diamondback moth (Plutella xylostella Linnaeus) and cabbage white butterfly (Pieris rapae L. Pieridae)  to the attacking insect(s) Fig. Although these studies demonstrate the potential for the exploitation of volatile compounds from basic to applied research, more studies are needed to decipher in-depth the concept of blends and their meaning for insect behaviors. Furthermore, future research also needs to focus on the effectiveness of the recruited natural enemies  in controlling specific insect pests, as well as the concentrations and active distances of emitted volatile blends.Metschnikowia gruessii, and M. reukaufii, attracts the parasitoid Aphidius ervi (Hymenoptera: Braconidae) via the production of volatile compounds  that associate and interact with different plant species and their organs Vorholt . These iPlants indirectly interact with the beneficial and symbiotic as well as pathogenic microbes that coexist with insects. A diverse community of microbes live within herbivorous insects and enter the plant during insect feeding. It has also been shown that insect-associated microbes can directly induce or suppress plant defenses after an herbivorous attack  levels from soil, alter critical ecological processes and thereby have significant consequences on multitrophic plant-mediated interactions. An increase in temperature can change the direct defenses such as primary and secondary metabolites. For example, in vineyards, an increase in temperature slows down plant defense through reduced production of anthocyanins and amino acids, but an increase in tannins in the grapes, thus making the plants more susceptible to insect damage   resulted in reduced affinity for volatiles produced during fermentation and gained an affinity for short-chain esters found in fresh fruits substrate, which could be considered as an ecological adaptation to a specific niche ) show the development of a well-structured olfactory memory that enables the ant to remember important olfactory landmarks. Further, the ants utilize the acquired knowledge to pinpoint their nest entrance in spatial scale  shows that both extrinsic and intrinsic factors drive host selection and preference. As reviewed by Takken and Verhulst , which could explain the evolution of preference and specialization for human odors  to oviposit. Several factors, including water vapor and VOCs, guide the gravid mosquito in making choices at increasingly fine scales  have been used successfully to disrupt mate location  recruited parasitoids and predators of aphids  plants was found to increase the number of visitations and predation of sentinel eggs by adult lady beetle, adult hoverflies, and predatory mites  was engineered in the laboratory to release Eβf, the alarm pheromone for many pest aphids  and increased the foraging by Aphidius ervi (Ervi), the natural enemy of these aphids  in vineyards  and Chilo partellus Swinhoe (Crambidae), and fall armyworm larvae, Spodoptera frugiperda (J.E. Smith) in maize crops  in pest management strategies against the codling moth in apple orchards and In 2020, the outbreak and rapid spread of novel coronavirus disease (COVID-19) broadly affected almost all aspects of human endeavor, including health, economics, social interactions, and education, among others. Education is a critical sector in modern society and has been primarily achieved best by teaching and training in a face-to-face environment. Following these lines, the ICE course for PhD students in chemical ecology and closely related research had enjoyed a face-to-face format annually for over a decade, while rotating between research facilities spread across three continents. Each year, both students and the selected trainers  would converge at one of the participating research facilities for the course activities. In 2020, the cycle was broken due to the unprecedented Covid-19 pandemic.What options do the ICE organizers and trainers have in responding and teaching the next generation of ICE scientists during a global pandemic, where social distancing is mandatory? In general, the advancement in technology has enabled the shifting of various human interactions from predominantly in-person to predominantly virtual. Such an unprecedented shift from the classical meeting in a host city or laboratory to cyber interactions during the Covid-19 pandemic became a new way to continue scientific exchange in an unprecedented time and has presented an opportunity for accelerated implementation of digital teaching and training technologies, and ‘unprecedented opportunities’ to ensure students do not miss out. Other positive aspects of using these technologies are reduced overall costs  and the inclusion of more students that otherwise could not participate due to either limited course capacity or else limited financial resources. Here, we suggest some ideas on how the ICE course can be successfully conducted during unprecedented situations and going forward even during more normal times.As pandemics such as Covid-19 demand restricted movement and social interactions, the first option to conduct ICE would be virtual-only attendance and teaching. This option offers real-time interactions over the internet. Several platforms for audio, videos, and chatting have been developed to enable interactive online learning experiences including lectures, discussion, and group training. Besides the virtual-only meeting, we suggest embracing the hybrid type of meeting where only students and trainers residing near the participating research facility can be allowed to participate in in-person and laboratory demonstrations. At the same time, those who would need to travel from far can follow the lectures and discussions virtually. The last option is pre-recording lectures and laboratory demonstrations. Here, the trainers can pre-record their lectures and demonstrations and have them streamed or played for the students and followed by a question-and-answer session with the particular lecturers. With this approach, copyright can be an issue. Therefore, we suggest the use of the non-downloadable format of the records or the lectures get played from a central platform and only for a limited time. While ICE courses are not just about teaching only, the students and trainers also get a chance to interact socially and talk about experiences with career progression. In all of the above-proposed methods of organizing and teaching the next ICE courses, we suggest the continued inclusion of social engagement activities even under a virtual setting, where both the trainers and students can interact and exchange from a social perspective, which is critical for students as early career scientists.Insects possess an inimitable capability to generate and recognize chemical cues and interact with a wide range of organisms in their environment. While these processes are only partly understood, the body of studies presented in this article demonstrates steady progress in understanding ICE and closely related fields. Indeed, ICE is a continuously growing and promising field of research where insights gained when studying these natural ecological interactions trigger new questions and sustainable contributions to the field and applications in agriculture and vector-borne disease control. Advancements in methods and instrumentations are accelerating manipulation of the molecular architecture and chemical events underlying signal emission, olfaction, and behavior. This is deepening our knowledge and helping to answer broader chemoecological and evolutionary questions, especially those linked to agriculture and disease vectors. Because ICE is an interdisciplinary field, the collaboration between professionals and early career scientists working in behavioral ecology, electrophysiology, analytical chemistry, molecular biology, data-driven science, medical entomology, and agroecology, among other disciplines, is vital. The ICE PhD course with its yearly organization rotating between SLU Alnarp in Sweden, PSU in the USA, MPI for Chemical ecology in Jena, Germany, and since 2019, ICIPE in Nairobi, Kenya, with the involvement of lecturers from other leading ICE research institutions, is a successful avenue for students to gain insights into the cutting edge of ICE research. Moreover, the course offers an opportunity to build bridges between disciplines, inspire new questions, and establish professional networks between participating students and lecturing scientists. We challenge the early career ICE scientists to follow and develop the present knowledge beyond their known borders.


WD repeat domain phosphoinositide-interacting protein 3 (WIPI3) is a member of the WIPI protein family, autophagy marker, that is associated with the malignant progression of various human cancers, but its role in hepatocellular carcinoma (HCC) is still unclear.Firstly, we collected the mRNA expression of WIPI3 in HCC through the platform of Oncomine, as well as the DNA copy number variations (CNVs), and verified it on human HCC cell line and the GEO database. Then, the subgroups and prognosis of HCC were performed by the UALCAN web tool. The mutation of WIPI3 was analyzed by cBioPortal. The coexpression of WIPI3 in HCC was identified from the LinkedOmics database, and function enrichment analysis was done using the LinkFinder module in LinkedOmics. Coexpression gene network was constructed through the STRING database, and the MCODE plug-in of which was used to build the gene modules; both of them were visualized by the Cytoscape software. Finally, the top modular genes in the same patient cohort were constructed through data mining in The Cancer Genome Atlas (TCGA) liver hepatocellular carcinoma (LIHC) by using the UCSC Xena browser.The results indicated that WIPI3 was frequently overexpressed in HCC, which could lead to a poor prognosis through the Kaplan–Meier (KM) analysis. Moreover, there existed mutations of WIPI3 in HCC, and the prognosis of WIPI3-altered group was significantly poor based on KM plotter data. Coexpression analysis showed that the coexpression gene of WIPI3 was associated with cell cycle and spliceosome. Further analysis suggested that WIPI3 and eukaryotic translation initiation factor 4A3 (EIF4A3) coordinately regulated the cancer cell cycle by spliceosome as a result of the strong positive correlation between them.In summary, WIPI3 is constantly overexpressed in HCC tissues, resulting in a poor prognosis; therefore, we can identify it as an effective target for the treatment of HCC. Hepatocellular carcinoma (HCC), with a high recurrence rate of 70%, is a leading cause of cancer-related mortality worldwide, as well as the most frequent liver cancer . Due to Autophagy is a highly conservative self-digestion process in eukaryotes and is crucial for cellular homeostasis, protein quality control, and pathogen defense , which oIn our work, we integrated the data of cancer gene expression from public database to perform multiple bioinformatics analyses, involving the comparison of WIPI3 expression between HCC and normal liver, detected the influences of WIPI3 mutation on HCC prognosis, and performed functional enrichment of WIPI3-related genes, revealing a new target for diagnosis and treatment of HCC.2.Human HCC cell line Huh-7 and normal liver cell line HL-7702 were conserved in our laboratory. Cells were cultured in Dulbecco’s modified eagle medium (DMEM) . The cell media contained 10% fetal bovine serum , 100 U/ml penicillin, and 100 mg/ml. Cells were maintained in a humidified incubator at 37°C with 5% CO® III 1st Strand cDNA Synthesis SuperMix for qPCR  and analyzed by Lightcycler 480 SYBR green I master supermix  incorporation in PCRs involving specific primers  according to the manufacturer’s instructions. First-strand complementary DNA was synthesized from equal amounts of total RNA (4 μg) using Hifair primers  and perf1, as well as the DNA copy number variations (CNVs) (P < 0.0001.We collected the mRNA expression of WIPI3 in HCC through the platform of Oncomines (CNVs) . Oncomins (CNVs) . The exp2 for external validation  through the LinkFinder module in the database, and the correlation of results was tested by the Pearson correlation coefficient. The pathway and network analyses of differentially expressed genes were performed by the LinkInterpreter module, the results of which were signed and ranked through the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways  is a public online databaseprojects . We usedhttp://string-db.org, and we screened out coexpressed genes with interaction scores greater than 0.4 to establish a protein–protein interactive (PPI) network  version 6.8, an online bioinformatics resource for investigators to comprehend the biological meaning behind the given list by the function of annotation, classification, and more . KEGG pat-test. All bar graphs are presented as SE, and p values less than 0.05 were considered significant.All statistical analyses were conducted using Prism7 (GraphPad Software). Differences were calculated using Student’s unpaired P < 0.01), as well as CNVs. We analyzed the transcription levels of WIPI3 in HCC tumors from a series of studies linked to the Oncomine database and found that the mRNA expression of WIPI3 in HCC tissues was obviously higher compared to normal tissues (P < 0.01), especially in 3–4 months (P < 0.0001)  curve, there was a close relationship between the expression of WIPI3 and the overall survival of HCC patients that the high expression of WIPI3 caused significantly poor overall survival  . As a reIn the TCGA database, we analyzed the coexpressed genes of WIPI3 in 371 LIHC cases through LinkedOmics . As showThe outcomes of Gene Ontology (GO) analysis carried out by GSEA in LinkedOmics indicated that differentially expressed genes correlated with WIPI3 were mainly located in the chromosomal region, spliceosomal complex, microtubule, transferase complex, nuclear chromatin, and ubiquitin ligase complex, where they primarily participated in DNA replication, organelle fission, mitotic cell cycle phase transition, and protein–DNA complex subunit organization. They acted as structural constituents in histone binding, ATPase activity, protein serine/threonine kinase activity, chromatin DNA binding, and tubulin binding . The funBy using the STRING database, the top 100 significantly coexpressed genes were built into a protein–protein network, and Cytoscape (MCODE plug-in) was used to establish the most important module . Based oUsing the UCSC Cancer Genomics Browser to hierarchically cluster the five hub genes with WIPI3, it was found that the expression pattern between WIPI3 and EIF4A3 gene was consistent . BesidesSubsequently, the expression of EIF4A3 at cancer stages and tumor grades in the TCGA database showed that EIF4A3 was overexpressed in HCC patients compared with normal people . We screin vitro. In this study, the expression of WIPI3 remained at a high level with the emergence and development of HCC, leading to poor prognosis, which suggested that WIPI3 could be identified as a potential indicator of HCC diagnosis. We also displayed the mutation of WIPI3 in HCC by cBioPortal. Although the frequency of WIPI3 was altered very low, the WIPI3 altered group displayed a poor prognosis in HCC. Finally, the coexpression genes related to WIPI3 in HCC were analyzed through the LinkedOmics database. GSEA function indicated that coexpression genes participated in cell cycle and spliceosome in GO and KEGG pathway analysis. EIF4A3, filtered from the coexpression genes, could be a partner of WIPI3 in HCC to regulate the development of HCC.WIPI3, a member of the WIPI protein family, is involved in cell autophagy pathway and lipid binding , but itsThe clinical diagnosis of HCC at an early stage is a severe challenge. At present, the most common indicator at early screening of HCC is alpha-fetoprotein (AFP), but not all HCC patients are AFP positive . Hence, Copy number variations may have important implications on genome, destroying genes and altering genetic content, causing phenotypic differences . In our The importance of WIPI3 in the pathogenesis of HCC is proved from the perspective of genomic stability and gene expression. Our study shows that the high expression of WIPI3 in HCC has a profound influence on genome stability as well as on multiple steps of gene expression  and cell cycle. We used a series of bioinformatics tools for neighbor gene analysis of tumor data from public databases. As a result, WIPI3 is specifically related to several spliceosome complex components .EIF4A3, due to its helicase activity in RNA , particiThe cooperative regulation of EIF4A3 and WIPI3 in cancer cells may indicate a special spliceosome mechanism that can synchronize rapid cell proliferation based on the maintenance of genomic stability. Cotargeting these two collaborative proteins might be an effective treatment for HCC. In conclusion, we have confirmed the upregulation of WIPI3 and its partner EIF4A3 in HCC and verified their importance as prognostic factors. Our research suggests that WIPI3 may be a promising molecular target for the diagnosis and treatment of HCC.This study systematically integrated public sequencing data to guide the research of WIPI3 in HCC. It was found that WIPI3 was highly expressed in HCC, which predicted a poor prognosis. The coexpressed gene of WIPI3 in HCC is closely associated with cell cycle and spliceosome, which causes WIPI3 mutations that are leading to lower mortality in HCC patients. Our work shows that EIF4A3 is an important partner of WIPI3 in HCC and suggests that WIPI3 may be an important potential biomarker for HCC.The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found in the article/QK and TW designed and conceived the research. TL did the bioinformatics analysis and wrote the manuscript. QS collected the datasets. ZD searched the literatures. All authors approved the final manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


This article explores a historical challenge for contemporary accounts of the role that the desire of being esteemed can play in exercising social control. According to Geoffrey Brennan and Philip Pettit, the economy of esteem normally has two aspects: it is supportive of virtuous action and it occurs spontaneously. The analysis of esteem presented by the 18th‐century materialist Claude‐Adrien Helvétius challenges the intuition that these two aspects go together unproblematically. This is so because, in Helvétius's view, the desire for esteem is always triggered by sensible interest. In the frequent situations where sensible interest diverges from the demands of virtue, the economy of esteem thus can be spontaneous but will not be supportive of virtue. Helvétius allows for cases of moral luck where sensible interest coincides with the demands of virtue but regards these cases as rare occurrences. This is why he believes that a functioning economy of esteem crucially depends on political constitutions—in which case the economy of esteem can be supportive of virtue but will not be spontaneous. This conception analyzes the quasi‐economic aspects of the role that the desire for esteem and the aversion against contempt has in motivating action: esteem can be understood as a scarce good; its scarcity induces competition; and even if esteem cannot be transferred in any direct way, one can provide a range of esteem services that enhance the esteem in which others are held. In their ground‐breaking work on the economy of esteem, Geoffrey Brennan and Philip Pettit make two claims that sound highly plausible, both regarded in themselves and read in combination with each other: (1) Unless in circumstances of secrecy and oppression, the economy of esteem is supportive of virtuous behavior since if persons act virtuously, “then they get the extra reward of being thought to be virtuous”  treats such situations as exceptions to the more frequent occurrence of situations where actions are visible and those experiencing their consequences are not dominated. And certainly, one easily can identify a multitude of everyday situations—think of the reasons that people do not free ride in public transportation—where such mechanisms lead to desirable consequences. Brennan and Pettit also point to a wealth of empirical research that documents the occurrence of behavior that creates trust and trustworthiness between agents in the absence of any institutional sanctions . The desire for mutual esteem could explain why these phenomena occur. Claim (2) explains what is so attractive about the economy of esteem as a mechanism of social regulation: If the competition for esteem ordinarily takes place without presupposing any institutional framework, and if the mere anticipation of being regarded unfavorably by others constitutes a powerful motivation for acting, then the economy of esteem can be regarded as an almost cost‐free mechanism of social coordination  or (2) in isolation but rather asks whether the  or (2) iHelvétius's central thesis is that esteem always reflects the tastes and interests of the subject who is passing judgment, be it an individual, a group, or a nation. On the basis of this thesis, he offers an analysis of pathologies of esteem that are not restricted to the cases of secrecy and oppression but are rather the outcome of the structure of sensible interest. I will explore these issues in Section 2De l'esprit (1757), Helvétius addresses two questions that are central to any treatment of esteem: (a) What is the object of esteem? (b) Why do we desire to be esteemed? Helvétius's answers to both questions derive from his account of sensible interest for human motivation. As to the first question, Helvétius claims that “personal interest is the unique and universal ground for appreciating the merit of human actions” . By contrast, what he calls “felt esteem” (estime sentie) depends on the conformity of ideas just described . In Helvétius's view, the conditions for felt esteem lead to irrational practices of esteem. One aspect could be described as a systematically distorted self‐esteem that results from the fact that our ideas have the highest degree of conformity with our own ideas. The high opinion that we have of ourselves and the admiration that we show for ourselves is not so much the result of pride than of the “necessity in which we are to esteem ourselves in preference over others” . Moreover, since no two humans have exactly similar ideas, we think that we think better than anyone else . This does not exclude the insight that one is much inferior to others in highly specialized skills; still, one believes in one's own superiority over specialists by denying the importance of their skills and by elevating the importance of common sense and social versatility . Another aspect of irrational practices of esteem shows itself in systematically distorted low esteem for others. One example that Helvétius mentions is the contempt between members of different social classes . Another example is the contempt for other historical ages . A further example . He understands this view as an alternative to the view that we desire the esteem of others because we are uncertain about our personal merits. In particular, he rejects the hypothesis that we seek a sentiment of our own excellence that we cannot get without the feedback from and comparison with others (ibid.). In support, he offers a group of arguments: (a) The first argument starts from a thought experiment: Imagine that there were other inhabited worlds and that some beings with super‐human powers were able to spread your fame in these other worlds. Would you prefer galactic esteem over national esteem? Helvétius suggests that everyone would prefer national esteem . As a related consideration concerning esteem in remote regions of our planet, he suggests this is so because our conationals (in contrast with inhabitants of other worlds and inhabitants of remote regions of our planet) can do something useful for us . (b) Humans show a desire for high political positions even under circumstances where it is politically impossible to achieve much. People have this desire even when they know that they will attract public contempt for their indifference toward public esteem. Hence, in situations where they can procure for themselves personal advantages without gaining esteem, they will prefer these advantages over being esteemed . (c) One is more flattered by esteem from the powerful than from those in low positions; hence, interest in being esteemed is proportional to the advantages that esteem can procure (ibid.). (d) Everyone in France prefers military fame to literary fame because the public gives more esteem to a general than to a philosopher . Similarly, almost everyone prefers the esteem of the unenlightened multitude over the esteem of some few enlightened persons because “the larger number of admirers recalls to our mind the enjoyable image of the pleasures that they can procure for us” . (e) Experience with oriental despotism shows that people show contempt for the striving for public esteem when the political system does not offer any rewards for merits but rather discourages actions that document moral and intellectual virtues . From these arguments, Helvétius concludes that the love of esteem reduces to the love of the pleasures arising from being esteemed .De l'homme (published posthumously in 1773), Helvétius returns to these issues and gives them a further edge by claiming that the desire for esteem is grounded in the desire for power. He maintains that the interest in happiness is nothing but an interest in having all the means necessary to be happy; and, as he argues, this interest boils down to an interest in power because power is the personal quality sufficient to procure the means necessary for happiness: “The love of power is founded on the love of happiness, and it therefore is the common object of all our desires; also, wealth, honor, fame … respect, justice, virtue … are in us nothing other than love for power disguised under different names” . This is why Helvétius takes the interest in esteem to be nothing but a disguise that the interest in power takes: “The one who spends the night under arms or in the office imagines loving esteem, but that's an error. Esteem is nothing but the name that one gives to the object of one's love, and the thing itself is power” .In heroic virtue. This is so because preferring death to humiliation—a virtue for which Helvétius has high respect . Helvétius's analysis of the connection between the desire for esteem and sensible interest thus cannot (perhaps with the exception of his treatment of heroic virtue) be dismissed as incoherent.If this line of argument is coherent, then it would become clear why the malfunctioning of the economy of esteem is inherent in psychological dynamics common to all humans. However, the coherence of Helvétius's argument has recently been questioned by Francesco Toto. As Toto maintains, there is an unresolved tension between Helvétius's view that our desire for esteem is always driven by sensible interest and Helvétius's view that virtuous citizens prefer esteem over wealth and sincere admiration over false flattery Toto, , p. 168?at the same time. This is so because he holds a spontaneous disposition toward seeking esteem through virtuous action to be a matter of moral luck. Consequently, he takes a widespread disposition toward seeking esteem through virtuous action to be a matter of political constitutions.Helvétius's point thus seems to be that, whether or not individuals seek esteem, the actual motivation behind their actions is their interest in pleasure and power. This view explains why the economy of esteem can be spontaneous, widespread, but still not conducive to virtue. Such situations occur whenever pleasure in the sameness of ideas arises from shared immoral prejudices and when public esteem procures power to those who act according to shared immoral prejudices. In this way, Helvétius's analysis goes beyond the analysis of the effects of the “intangible backhand” offered by Brennan and Pettit since it offers an explanation of why the malfunctioning of the economy of esteem need not be restricted to situations of oppression and secrecy. Still, as we shall see in the following two sections, Helvétius also allows for the possibility that individuals are spontaneously motivated to fulfill duties of morality, as well as for the possibility that the economy of esteem widely supports virtuous action. The real challenge that he poses for contemporary accounts of the economy of esteem is his insight that the desire for esteem may not support morality spontaneously and in a widespread manner 3But what influence can a difference in instruction occasioned by some light difference in surrounding objects have on minds? Well, does one ignore that a small number of different ideas that are combined with ideas that two humans already have in common, can produce a difference in their entire way of seeing and judging? From a contemporary perspective, Helvétius's conception of moral luck could be described as a conception of “constitutive” luck—that is, of luck that explains what an individual is like. Helvétius's remarks about the differences in education also make it clear that the comparison that he has in mind is nothing as sophisticated (and problematic) as a comparison between an individual in the actual world and individuals in nearby possible worlds,As to the role of luck in our moral lives, Helvétius proposes: “Without despising the vicious man, one must mourn him, and congratulate oneself for a lucky natural constitution …” . Moral luck here is characterized as a kind of natural luck—luck that has brought forth natural dispositions that others are lacking. Among these natural dispositions, Helvétius counts intellectual capacities that are the result of chance in education: “[C]hance has a greater part in our education than we think. It is chance that puts certain objects before our eyes and consequently occasions for us lucky ideas and sometimes leads us to greater discoveries” ibid., 2:7). Accordingly, he draws attention to the fact that no one receives exactly the same education because circumstances never coincide in two persons . Helvétius never gives any definition of the concept of chance. But he observes that a vast amount of small impressions goes into education in early age, most of which are entirely unintended by the educators . Some of them are the unintended side effects of intentional actions, such as when a child pays attention to the shades of color of pots on the window board while being locked up in a room as a form of punishment . It is the effects that such forgotten small impressions have on conscious ideas that Helvétius describes as the effects of chance::7. AccorStill, describing Helvétius's conception of moral luck in this way seems to make it vulnerable against a recent criticism of the concept of constitutive luck. As David Enoch and Andrei Marmor argue, if character traits cannot be regarded as the object of intentional control, then individuals could not be thought worthy of praise or blame for such character traits , Helvétius there remarks: “These rights oppose themselves to the despotism of a power that is entirely foreign to primordial convention and to the fundamental laws of the constitution of societies” It is not only the light of reason, nor only the inner sentiment of the law alone that guides humans in their deliberations. Inner sentiment makes them attentive; reason enlightens them; but in addition, it is necessary that sensations represent the objects that provide the motivations of physical goods or evils, of moral goods or evils, which determine the decisive acts of the will. This commitment to a non‐theological version of natural law theory is highly relevant for Helvétius's views on the goals of philosophy and politics. As he suggests, the task of the philosophers is to think about the morality relating to legislation, and the task of the legislator is to find out what, under ever changing circumstances, is useful for a given community . “But the one and the other grounds the institution of public laws on human nature and on the law of laws, and on the physical dynamics of human actions, and on justice that is co‐essential for the general well‐being of society” . The philosopher and the legislator who fulfil this task thus exemplify the character traits that give substance to the idea of constitutive moral luck. Because these character traits are genuinely useful both for the community and for those who have these traits, they are a legitimate object of positive moral evaluation, even if they are not the outcome of intentional control.Helvétius is not usually considered to be one of the early modern natural law thinkers; however, as Sophie Audidière and Ann Thomson have emphasized, in a postscript to Interest presents objects to us only from the side that it is useful for us to apperceive them. When one judges according to public interest, it is not so much due to the justness of one's mind or the justice of one's character that one should render honor than to the chance that has placed one in circumstances where we have an interest to see as the public sees. When actions are judged according to public interest, Helvétius holds that those who pass such judgments fulfill nothing other than “the passion that an enlightened pride gives them for virtue and, consequently, nothing other than obeying, like every other society, the law of personal interest” . Even in such cases, “interest is the only distributor of esteem” . More specifically, “he justice of our judgments and our actions is never anything but the lucky coincidence of our interest with the public interest” . Consequently, Helvétius holds that “every author who gives new ideas to the public can hope for the esteem of only two kinds of humans: either young people who, because they have not yet adopted any options, still have the desire and the leisure to instruct themselves; or those whose mind, being a friend of truth and analogous to the mind of the author, already suspects the existence of the ideas that he presents to him” .The morally lucky therefore can develop mutual esteem that is in accordance with the demands of morality. Helvétius describes our readiness to judge the actions and ideas of others according to the norms of morality as the outcome of a multiplicity of accidental factors:For Helvétius, esteeming ideas that one recognizes as being superior to one's own thus is a genuine possibility.4Moral luck, for Helvétius, thus explains why practices of mutual esteem that are both spontaneous and supportive of virtue can occur. At the same time, it explains why such practices occur only in exceptional cases. In this sense, his account of moral luck does not explain much about our ordinary practices of esteem. This is exactly what makes his considerations concerning moral luck so interesting from the perspective of the contemporary debate concerning the economy of esteem: While Brennan and Pettit believe that a morally beneficial economy of esteem occurs both widely and spontaneously, Helvétius's considerations concerning moral luck raise the suspicion that cases of practices of mutual esteem that are both beneficial and spontaneous may not be widespread.political task .What is more, Helvétius's conception of moral luck explains the presence of a claim that has been noted by his interpreters but that remains puzzling if read independently of the issue of moral luck. The claim that I have in mind is Helvétius's thesis that the “administration of the monies of honors” is mainly a These interpretive puzzles dissolve once one takes the issues of sensible interest and moral luck into account. The very concept of luck implies that, due to the nature of chance, most individuals do not fall into the category of the morally lucky. The crucial consequence that Helvétius draws from the rare occurrence of moral luck and the frequent distortion of our judgment through sensible interest is that it is the task of legislators to force subjects to observe the law of nature . Helvétius regards this as a realistic project because morality is “bound to our well‐understood interest” . As a remedy for destruction of societies, he recommends “to interest individuals, through good laws, to concur to the general well‐being of the society in which they live” . Helvétius claims that his conception of moral motivation does not exclude freedom. As he argues, even the insight into the rewards and punishments that the legislator has attached to laws and their transgression forms part of rational deliberation that leads to choosing the course of action that is most strongly in the personal interest . But clearly, even if there may be a sense in which actions guided by rewards and punishments can be regarded as free, they cease to be spontaneous.politics but the result of specific political constitutions. It is here that the connection between the desire for esteem and sensible interest becomes crucial. Helvétius acknowledges that the desire for esteem can motivate the search for truth; but he notes that the love for the true is always subordinate to the love of happiness. In his view, it is impossible to love in the true anything other than the means for increasing happiness. Hence, few will search for truth under a government where truth is despised . He also offers an argument for why the economy of esteem in a despotic regime cannot get off the ground:[I]f honors derive their price from the way in which they are administered, and if in the Orient sultans are the distributors of honor, one senses that they must often discredit honors through the bad choice of those whom they decorate with them. Also, in those countries, honors … cannot vividly flatter pride because they are rarely united with glory, which is not at the disposal of princes but of peoples; for glory is nothing other than the expression of public recognition. Generally, Helvétius holds that under a government that debases talents, ingenious persons are entirely the result of chance or of the memory that one has preserved of the honor that has been bestowed on virtue and talent in the past . By contrast, under a government that honors merit, love for fame is not the result of chance but of the constitution of the state . Therefore, he takes it to be a political task to trigger the love for public esteem . But it is a task that can be accomplished only in particular political constitutions: “Why is fame regarded as a plant of republican soil, which, degenerated in despotic countries, never grows there with a certain strength? It is because in fame one loves properly only power, and in an arbitrary government, all power disappears in the face of the power of the despot” . On the contrary, “[i]n a free nation, public reputation and esteem is a power, and the desire for this esteem there becomes consequently a powerful principle of action” . But even in a republican constitution, the economy of esteem always is a fragile interplay between political agency and public opinion. This is why those who are active in defining positions of honor have to develop a fine discernment for when a certain kind of honor has become too common since “honors owe their price only to the opinion of humans” . Since this is such a delicate task, Helvétius proposes the creation of institutions that have the task of regulating the distribution of honors in a society (ibid.). Under a republican constitution, the desire for esteem thus in fact plays a central role in social regulation; if the political institutions define positions of honor in such a way that personal and public interests are united, the desire for esteem also may be largely supportive of virtuous action; but the fact that esteem leads to enough power to secure sensible interest may be shaped more profoundly by political institutions than we may be aware of.At the same time, the virtue‐supportive function of the economy of esteem, in Helvétius's view, is not just result of 5How plausible, from an empirical point of view, is Helvétius's view that the economy of esteem is, to a large degree, an outcome of political institutions? This question is pressing in the light of extensive psychological research that documents the extent to which human judgments succumb to a variety of biases. Why should we think that the esteem in which persons hold themselves and others could be more strongly influenced by a structure of institutionalized rewards and sanctions than by prejudices that are favorable for their self‐esteem? Let me focus here on the example of national biases, which play a significant role both in contemporary psychological research and in Helvétius's moral philosophy.lower levels of subjective well‐being , while identification of type (2) is correlated with higher levels of subjective well‐being This folly, common to all nations, … leads them not only to disdain the ways of living and the customs different from their own, but also leads them to regard the superiority that some have over others as a gift of nature—a superiority that they owe only to the political constitution of their state. Accordingly, he holds that “contempt for a nation is always an unjust contempt” . But no matter how unjust contempt between nations may be, he nevertheless describes it as being grounded in human nature . That the objects of our esteem and the motivation for seeking esteem are determined by sensible interest, he notes, can also explain why personal interests deriving from laziness and fear can lead us to esteem cruelties in one's own nation that one despises in other nations. For instance, the coexistence of widespread contempt for the ritual practices of Maya priests with widespread esteem for the cruel methods of the Inquisition is a vivid illustration for how much interest can distort esteem for other nations . In addition to the interests deriving from laziness and fear, Helvétius also identifies the interest of vanity to explain national biases: “Each nation will always exalt the virtues that it owes to the form of its government to a gift of nature. The interest of vanity advises it do so: and who could resist the advice of interest?” .Helvétius, too, distinguishes between two kinds of national identification, one that involves unjustified feelings of national superiority and one that involves an appreciation of political institutions that unite personal and public interests. As to the first kind of national identification, Helvétius is clear about the pervasive occurrence of unjustified national biases. One of his explanations for the occurrence of national biases invokes the sensible interest that we have in esteeming only ideas that are similar to our own ideas. As he argues, this psychological mechanism explains why one esteems or disesteems the ideas of other societies according to their correspondence with or divergence from one's own ideas . If a political constitution unites personal and public interests, among other things through the rewards connected with merit, it is easy to see how sensible interests are served by identifying with this constitution. This is why Helvétius holds that republican constitutions lead to civic spirit (l'esprit de citoyen), while monarchic constitutions lead to an indifference toward public affairs .However, conceptualizing the persistence of national biases in the framework of sensible interests also opens up a plausible conception of how such biases could be overcome. What is needed are the workings of Helvétius's view that national identification connected with an appreciation of one's own political institutions could serve the interest in self‐esteem better than entertaining national biases is supported by the psychology of nationalism. The empirical findings mentioned above indicate that national identification based on feelings of superiority and derogation of other nations actually serves very poorly the interests of those who take this attitude. This is why it is not unrealistic for a political constitution that uses the desire for esteem to connect what is personally useful with what is publicly useful will create a mentality that does not try to derive self‐esteem from national bias but rather from actions that contribute to the well‐being of the community and that is perceived and rewarded as such. And, as we have seen, there is empirical support for the assumption that national identification based on a positive evaluation of national institutions and values in fact increases subjective well‐being. All that Helvétius would add to this empirical finding is the reminder that such a form of patriotism presupposes political constitutions that really unite private and public interests.6spontaneously, this is mainly a matter of moral luck. The contemporary discourse about moral luck is almost completely separated from the discourse about the economy esteem. The only exception to this is a brief but thought‐provoking article by Jeffrey Fry. As Fry points out, there are two kinds of situations where significant others can have a lucky influence on self‐esteem: personal interactions in early childhood that give growth to self‐esteem and personal encounters that help to recover self‐esteem independently of being successful in competitions. Fry argues that such situations could be described as involving “grace” rather than “chance”: while chance implies the absence of intentional action, interactions of the type described are intentional and the proper object of thankfulness . But he presses the analysis of such actions one step further by asking about the conditions on which they depend, and some of these conditions—physiology and the long history of personal experiences—inevitably involve factors that are not intended outcomes of actions. This is why Helvétius sees a close connection between the virtue‐supportive function of the economy of esteem and moral luck. In his view, persons with lucky dispositions can develop esteem for virtuous actions, as well as the desire to be esteemed for virtuous actions, just because this is what turns out to be pleasurable for them.What is interesting about Helvétius's considerations concerning the desire for esteem is that they envisage a possibility that, as far as I can see, has not been discussed in the contemporary debate about the economy of esteem: the possibility that, if the desire for esteem supports virtuous action ess Fry, . As we hDue to the role of chance, such situations will occur rarely. In the absence of political steering, Helvétius therefore argues that personal interests will lead most individuals to systematically flawed practices of esteem—ranging from inflated self‐esteem, to derogation of others, to contempt for other nations. This is why he holds that political arrangements are crucial for the proper working of the economy of esteem: the distribution of esteem must be regulated in such a way that esteem remains a scarce good; it must be regulated by criteria of justice that are sensitive to the usefulness and difficulty of certain activities; and the rewards for esteem must be regulated in such a way that esteem gives enough power to fulfill sensible interests. Helvétius also offers an analysis of what happens to the economy of esteem in situations of oppression. What he has in mind is not the observation that the members of the oppressing group will not be concerned about the esteem of the oppressed. Rather, he notes that despotic regimes annihilate the economy of esteem even within the oppressing group. As he argues, this is so because under despotic rule, esteem is no guarantee for power and, hence, no guarantee for sensible pleasure. People know this and, except for the morally lucky, will not be motivated by a desire for esteem.civic behavior—hence, behavior that takes place in the context of specific political constitutions (ibid.).An observation that Brennan and Pettit adduce to support their spontaneity thesis in fact offers further empirical support for Helvétius's contrary thesis. As Brennan and Pettit observe, the failures of free markets in post‐communist countries show that fair market practices cannot occur without a tradition of civic values . The divergence between Helvétius and Brennan and Pettit thus does not concern the feasibility of institutional influences on the economy of esteem. Rather, it concerns the question of priority: While for Brennan and Pettit, the existence of widespread, reliable, and virtue‐supporting practices of mutual esteem and disesteem is the basis upon which subsequent institutional design and redesign depends, Helvétius holds that the existence of widespread, reliable, and virtue‐supporting practices of mutual esteem and disesteem depends upon institutional design.Helvétius's challenge could thus be wrapped up as follows: (a) As long as our personal interests are not in line with the duties of morality, our practices of esteem will be systematically flawed. Such practices may be spontaneous and widespread, but they do not contribute to virtuous action. (b) An economy of esteem that is spontaneous and supportive of virtue is a genuine possibility, but due to its dependence on moral and intellectual luck, it will be a rare occurrence. (c) An economy of esteem that is widespread and supportive of virtue is a genuine possibility, but moral luck will have to be supplemented by political planning and education that guarantees that virtuous action is rewarded with an amount of power that is sufficient for fulfilling sensible interest. In such cases, the economy of esteem will be spontaneous only to a small extent. If there is something to these three considerations, then the connection between the claim that the economy of esteem is usually supportive of virtue and the claim that it is mainly a spontaneous phenomenon may be more problematic than is obvious on first sight. This is a challenge that still deserves a response.


Tyrosinase-related protein 1 (TYRP1) is one of the three human melanogenic enzymes involved in the biosynthesis of melanin, a pigment responsible for the color of the skin, hair, and eyes. It shares high sequence identity with tyrosinase, but has two zinc ions in its active site rather than two copper ions as in tyrosinase. Typical tyrosinase inhibitors do not directly coordinate to the zinc ions of TYRP1. Here, we show, from an X-ray crystal structure determination, that phenylthiourea, a highly potent tyrosinase inhibitor, does neither coordinate the active site zinc ions, but binds differently from other structurally characterized TYRP1-inhibitor complexes. Its aromatic ring is directed outwards from the active site, apparently as a result from the absence of polar oxygen substituents that can take the position of water molecules bound in the active site. The compound binds via hydrophobic interactions, thereby blocking substrate access to the active site. The human melanogenic enzymes tyrosinase (TYR), tyrosinase-related protein 1 (TYRP1), and tyrosinase-related protein 2 (TYRP2) take part in the biosynthesis of melanin, a pigment that is responsible for the color of the skin, hair, and eyes . The preAlthough the role of TYR in the melanin biosynthetic pathway is generally accepted, the function of TYRP1 is less clear . BecauseB. megaterium tyrosinase [Binding of L-tyrosine and the L-DOPA analogue L-mimosine to TYRP1 showed that these compounds do not directly interact with the zinc ions of TYRP1, in contrast to, for example, the tyrosine analogue tyrosol, which directly coordinates Cu-A in rosinase . To furtrosinase . Our resN-glycosylation patterns. Three N-glycosylation sequons ) are conserved in the human melanogenic enzymes. No disease-related mutations are known for TYR Asn86 and Asn290, suggesting that glycosylation of these sites is not essential for functionality. In contrast, Asn371 mutations have been found in patients with classic tyrosinase-dependent oculocutaneous albinism. Asn371 is located at the rim of the entrance to the active site pocket, about 15–20 Å from the TYR copper ions, suggesting that the absence of glycosylation at this site severely compromises the enzyme’s activity, or stability, or both.TYR, TYRP1, and TYRP2 not only differ in their enzymatic activity, but also in their Various compounds with well-known reductive tyrosinase activity were tested in an on-plate colorimetric assay for their potency against the melanosomal domain of TYR. Interestingly, these experiments demonstrated that PTU is the most potent inhibitor of human tyrosinase , similarThe crystal structure of TYRP1-3M with bound PTU was elucidated at 2.2 Å resolution ; Table 1Superimposition of the PTU-inhibited TYRP1-3M structure and TYRP1-3M structures with bound kojic acid, mimosine, and tropolone  revealeThe precise function and activity of TYRP1 has been puzzling . InitialIpomoea batatas catechol oxidase  was the most potent inhibitor of human tyrosinase . It bindry 1BUG, ), anothe2+ and Zn2+ ions are very similar  affects the binding mode of PTU. The ionic radii of Cuectively ) and alsSuperimposing the TYRP1-3M PTU structure on the human TYR model  PEG 3000, with a protein to reservoir ratio of 3:1. Rod-like crystals appeared in 2 days and grew to full size in 7 days. For soaking PTU in the crystals, native crystals were fished in a cryo drop and soaked with reservoir solution supplemented with 20% (v/v) glycerol and saturated PTU for 5 to 10 min before flash freezing in liquid nitrogen for diffraction experiments.TYRP1-3M native crystals were generated in a condition containing 0.1 M Tris (pH 7.0), 0.2 M NaCl, and 30% (X-ray diffraction data collection was carried out at the MASSIF-1 automatic beamline of the European Synchrotron (ESRF), Grenoble, France . The datThe full amino acid sequence of human tyrosinase  was submitted to the Swiss-model server . Four mo


Stephania cepharantha Hayata (Menispermaceae), has a range of therapeutic potential in clinical conditions. Whether it affects the activity of human liver cytochrome P450 (CYP) enzymes remains unclear. Cepharanthine (CEP) extracted from the roots of in vitro using human liver microsomes (HLMs) with specific probe actions and probe substrates. In addition, the enzyme kinetic parameters were calculated. The effects of CEP (100 μM) on eight human liver CYP isoforms  were investigated 50 values of 16.29, 25.62 and 24.57 μM, respectively, but other CYP isoforms were not affected. Enzyme kinetic studies showed that CEP was not only a non-competitive inhibitor of CYP3A4 but also a competitive inhibitor of CYP2E1 and CYP2C9, with Ki values of 8.12, 11.78 and 13.06 μM, respectively. Additionally, CEP is a time-dependent inhibitor for CYP3A4 with IK/inactK value of 10.84/0.058 min/μM. The results showed that the activity of CYP3A4, CYP2E1 and CYP2C9 was inhibited by CEP, with ICin vitro studies of CEP with CYP isoforms indicate that CEP has the potential to cause pharmacokinetic drug interactions with other co-administered drugs metabolized by CYP3A4, CYP2E1 and CYP2C9. Further clinical studies are needed to evaluate the significance of this interaction. The  Stephania cepharantha Hayata (Menispermaceae), known to have anti-inflammatory and immunomodulatory activities  is a biscoclaurine alkaloid, extracted from the roots of Cytochrome P450 (CYP) enzymes are membrane-bound hemoproteins, that play a vital role in the biotransformation of xenobiotics, including drugs, environmental pollutants, carcinogens and endogenous substrates , testosterone (CYP3A4), coumarin (CYP2A6), chlorzoxazone (CYP2E1), dextromethorphan (CYP2D6), diclofenac (CYP2C9), S-mephenytoin (CYP2C19), and paclitaxel (CYP2C8) were used as probe substrates to determine the effects of CEP on eight CYP enzymes. In addition, enzyme kinetic studies were conducted to determine the inhibition mode of CEP on CYP enzymes.CEP has been widely used in clinic; however, the effect of CEP on the activity of CYP enzymes was unclear. In this study, the effect of CEP on eight major CYP isoforms in human liver microsomes (HLMs) was investigated. The effects of CEP on the activity of CYP enzymes were investigated in HLMs. Through comparing the probe actions in the presence or absence of CEP, the activity of eight CYPs enzymes was obtained. The enzyme kinetic parameters were also calculated to quantify the effect of CEP. With the help of HPLC, the probe substrates were analysed.D-Glucose-6-phosphate, glucose-6-phosphate dehydrogenase, corticosterone (≥98%), NADP+, phenacetin (≥98%), acetaminophen (≥98%), 4-hydroxymephenytoin (≥98%), 7-hydroxycoumarin (≥98%), 4′-hydroxydiclofenac (≥98%), sulfaphenazole (≥98%), quinidine (≥98%), tranylcypromine (≥98%), chlorzoxazone (≥98%), 6-hydroxychlorzoxazone (≥98%), paclitaxel (≥98%), 6β-hydroxytestosterone (≥98%), clomethiazole (≥98%) and furafylline (≥98%) were obtained from Sigma Chemical Co. Montelukast (≥98%) was obtained from Beijing Aleznova Pharmaceutical . Coumarin (≥98%), diclofenac (≥98%), dextromethorphan (≥98%) and ketoconazole (≥98%) were purchased from ICN Biomedicals . Pooled HLMs were purchased from BD Biosciences Discovery Labware . All other reagents and solvents were of analytical reagent grade.CEP (≥98%) and testosterone (≥98%) were obtained from the National Institute for the Control of Pharmaceutical and Biological Products . The chemical structure of CEP is shown in O-deethylation for CYP1A2, testosterone 6β-hydroxylation for CYP3A4, coumarin 7-hydroxylation for CYP2A6, chlorzoxazone 6-hydroxylation for CYP2E1, dextromethorphan O-demethylation for CYP2D6, diclofenac 4′-hydroxylation for CYP2C9, S-mephenytoin 4-hydroxylation for CYP2C19 and paclitaxel 6α-hydroxylation for CYP2C8. Marker reactions of eight CYP isoforms were examined, and apparent Km values were estimated to characterize the pooled HLMs. All incubations were performed in triplicate, and the mean values were utilized. The typical incubation systems contained 100 mM potassium phosphate buffer (pH 7.4), NADPH generating system , the appropriate concentration of HLMs, a corresponding probe substrate and hispidulin (or positive inhibitor for different probe reactions) in a final volume of 200 μL.As shown in The concentration of CEP was 100 μM, and the positive inhibitor concentrations were as follows: 10 μM furafylline for CYP1A2, 1 μM ketoconazole for CYP3A4, 10 μM tranylcypromine for CYP2A6, 50 μM clomethiazole for CYP2E1, 10 μM quinidine for CYP2D6, 10 μM sulfaphenazole for CYP2C9, 50 μM tranylcypromine for CYP2C19 and 5 μM montelukast for CYP2C8. Probe substrates, positive inhibitors (except for dextromethorphan and quinidine which were dissolved in water) and CEP were dissolved in methanol, with a final concentration of 1% (v/v), and 1% neat methanol was added to the incubations without inhibitor. The final microsomal protein concentration and incubation times for the different probe reactions are shown in 50IC). Ki values were obtained by incubating various concentrations of different probe substrates  in the presence of 0–50 μM CEP.CEP (100 μM) was used to initially screen for its direct inhibitory effects towards different human CYP isoforms. For the CYP isoforms whose activities were strongly inhibited, secondary studies were performed to obtain the half inhibition concentration  was pre-incubated with HLMs (1 mg/mL) in the presence of an NADPH-generating system for 30 min at 37 °C. After incubation, an aliquot (20 μL) was transferred to another incubation tube  containing an NADPH-generating system and probe substrates whose final concentrations were approximate to IK and inactk values for the inactivation of CYP3A4, the incubations were conducted using higher probe substrate concentrations  and various concentrations of CEP (0–50 μM) after different preincubation times (0–30 min), with a two-step incubation scheme, as described above.To determine the versus the inverse velocity (Lineweaver-Burk plots), and the mean values were used to calculate maxV and mK. Inhibition data from the experiments that were conducted using multiple compound concentrations were represented by Dixon plots, and inhibition constant (iK) values were calculated using non-linear regression according to the following equation:iK is the inhibition constant, S is the concentration of the substrate, and mK is the substrate concentration at half the maximum velocity (Vmax) of the reaction. The mechanism of the inhibition was inspected using the Lineweaver–Burk plots and the enzyme inhibition models. The data comparison was performed using Student’s t-test and performed using IBM SPSS statistics version 20 .The enzyme kinetic parameters for the probe reaction were estimated from the best fit line, using least-squares linear regression of the inverse substrate concentration 50IC values of 16.29, 25.62 and 24.57 μM, respectively.The effects of CEP on the activity of CYP enzymes were investigated, through the probe reaction assays. Specific inhibitors of CYP1A2, CYP3A4, CYP2A6, CYP2E1, CYP2D6, CYP2C9, CYP2C19, and CYP2C8 were used as positive controls. As shown in Ki values of CEP on CYP3A4   over the in vivo concentration of the inhibitor  were approximately 0.05 μg/mL (Jiang et al. Ki and 50IC values of CEP. Consequently, the drug–drug interaction might not occur when CEP was co-administered with the substrates of the CYP3A4, CYP2E1 and CYP2C9. Considering the pharmacokinetic differences of humans and rats, further in vivo system studies are needed to identify the interactions of CEP with CYP isoform in humans.According to previous studies about the pharmacokinetic profiles of CEP in rats, the plasma in vitro and in vivo.However, there are also some limitations to this study. We found the inhibitory effect of CEP on the activity of CYP3A4, 2E1 and 2C9 and the kinetic characteristics were also obtained. The potential mechanism requires additional investigation. Therefore, the inhibitory mechanism underlying effects of CEP on the activity of CYPs needs more attention. On the other hand, during the biotransformation of drugs, UDP-glucuronosyltransferases is also involved. In fact, the effect of CEP on the activity of UDP-glucuronosyltransferases had not been studied. In future research, the effects of CEP on the activity of UDP-glucuronosyltransferases should also be investigated The effect of CEP on the activity of CYP enzymes was investigated in this article. CEP inhibited the activity of CYP3A4, and competitively inhibited CYP2E1 and CYP2C9, while other CYP enzymes were not be influenced. Therefore, CEP should be used carefully together with the drugs metabolized by CYP3A4, CYP2E1 and CYP2C9.


Arising from P. A. Tuckett et al., Nature Communications 10.1038/s41467-019-12039-2 (2019).1 report on short-term events of ice flow acceleration on five outlet glaciers of the northern Antarctic Peninsula and their relation to numerical model output of surface melt. The authors argue that the delivery of surface meltwater to the glacier bed transiently increases the basal water pressure and enhances basal motion, causing near-instantaneous flow acceleration followed by subsequent drainage causing deceleration. An outdated version of the grounding line (GL) vector, separating grounded and floating glacier ice, is used so that the majority of the analyzed velocity points are located on floating sections of glacier termini where meltwater drainage does not have any effect on subglacial water pressure. Our independent analysis of glacier surface velocities exhibits for the speed-up events only marginal changes in velocity on grounded ice and a significant increase of velocity on floating glacier sections progressing on prefrontal sea ice and ice mélange, clear evidence for the dominant influence of ocean dynamic forcing as previously reported by refs. 4.Tuckett et al.1 are time series of 6-day mean velocities between October 2016 and April 2018 for glaciers discharging into the Larsen A embayment , the Larsen B embayment , and the Gerlache Strait (Cayley Glacier). Velocities are derived from 6-day repeat-pass radar satellite images of the Sentinel-1 mission, aggregated into 1 km squared boxes at distances 1 km up to 10 km upstream of the glacier fronts. The interpretation of the velocity data in terms of ice flow dynamics is critically based on the assumption that the boxes are located on grounded ice. However, high-resolution digital elevation models (DEMs) of the TanDEM-X satellite mission, acquired in mid-2011, −2013, −2016 and the analysis of surface elevation change (SEC) show that major sections were afloat already in these years4. The authors used an outdated GL version, so that in total 24 out of the 36 velocity boxes shown are located on floating sections of glacier termini: all 10 velocity boxes on Hektoria Glacier, all six velocity boxes on Crane Glacier, three boxes on both Drygalski and Jorum glaciers and two boxes on Cayley Glacier.The main data products used in ref. 2. The persistent sea ice cover since mid-2011 in the proglacial bay impeded calving, leading to terminus advance whilst glacier thinning on grounded ice continued. Whereas the termini of Hektoria and Green glaciers extend into a wide bay, the tongues of the other studied glaciers are narrower and laterally confined by mountain ridges. The floating parts of these glaciers extend several kilometers inland of the front along the center of the terminus, whereas the lateral margins rest on slopes, as shown in Fig. In Fig. 5. Here, we argue that the velocity time series provide clear evidence for the dominant impact of oceanic forcing during the acceleration events highlighted by ref. 1. The examples in Fig. 6. Over the same time span, the young sea ice in front of the multiyear pack ice in the Larsen B embayment drifted eastward by 15 km. Oceanic processes also have a large impact on multiannual variations of flow velocity and mass balance. Persistent multiannual sea ice in the Larsen A embayment (from mid-2013 to 2016) and in the Larsen B embayment (since winter 2011) caused major decrease of flow velocities and the mass losses of grounded ice dropped from 9.73 Gt a−1 during 2011–2013 to 4.70 Gt a−1 during 2013–20164.We generated ice velocity maps of Antarctic outlet glaciers since 2015, in 12- and 6-day time intervals, depending on data availability according to the Sentinel-1 operation plan1 as a basis for inferring conclusions about the sources for speed-up. First, it is unclear which criteria are used for selecting the melt-induced acceleration events out of the full sample of velocity spikes coinciding with modeled surface melt. Second, the authors claim that a larger relative increase in velocity closer to the glacier front would be needed if marine processes were the trigger for the speed-up. Beside this claim being based on relative velocity change, whereas any flux and mass considerations must be based on actual velocities, no quantitative analysis is provided to support this statement. In addition, the assumption that surface meltwater of the transient events finds its way to the glacier bed and causes an increase of subglacial water pressure is speculative and not relevant for floating ice. Apart from melt intensity, the freezing state of the snow/firn/ice body needs to be taken into account. The Larsen outlet glaciers have cold snow/ice bodies in which a substantial portion of the meltwater released at the surface would freeze, in particular after cold periods7. Estimates on the intensity and spatial extent of melt events can be deduced from C-band backscatter signatures8. For example, the event in March 2018 shows a modest decrease of the backscatter intensity on only one date (27 March 2018), indicating a short period of modest surface melt not able to release sufficient water for drainage to the glacier bed.A further point of concern is the use of relative changes in velocity in ref. Another critical issue is the neglect of biases in the retrieved velocities caused by shifts in the radar line-of-sight (LOS) distance by several meters owing to changes in radar signal penetration associated with change from dry to wet snow and vice versa. Depending on the flow direction relative to LOS, this shift causes an underestimation or overestimation of the velocity (Supplementary Note The issues addressed above question the interpretation of the presented material in terms of melt-induced acceleration. Our analysis of velocity time series on grounded and floating glacier sections and the motion of proglacial sea ice and ice mélange confirm the findings of previous publications that changes in velocity and ice export of northern Antarctic Peninsula outlet glaciers during recent years have been primarily governed by frontal stress perturbations propagating up-glacier and by variations in oceanic boundary conditions.Supplementary InformationDescription of Additional Supplementary FilesSupplementary Data 1Supplementary Data 2


The present study aimed at examining the inhibitory effect of two atypical neuroleptics iloperidone and lurasidone on the main human cytochrome P450 (CYP) enzymes in pooled human liver microsomes and cDNA-expressed CYP enzymes (supersomes).N-demethylation/CYP1A2, diclofenac 4′-hydroxylation/CYP2C9, perazine N-demethylation/CYP2C19, bufuralol 1′-hydroxylation/CYP2D6 and testosterone 6β-hydroxylation/CYP3A4, respectively, using HPLC.The activity of these enzymes was determined by the following CYP-specific reactions: caffeine 3-Ki = 0.38 and 0.3 µM in liver microsomes and supersomes, respectively) and CYP2D6 via a competitive mechanism (Ki = 2.9 and 10 µM in microsomes and supersomes). Moreover, iloperidone attenuated the activity of CYP1A2 (Ki = 45 and 31 µM in microsomes and supersomes) and CYP2C19 via a mixed mechanism (Ki = 6.5 and 32 µM in microsomes and supersomes) but did not affect CYP2C9. Lurasidone moderately inhibited CYP1A2 (Ki = 12.6 and 15.5 µM in microsomes and supersomes), CYP2C9 (Ki = 18 and 3.5 µM in microsomes and supersomes) and CYP2C19 via a mixed mechanism (Ki = 18 and 18.4 µM in microsomes and supersomes), and CYP3A4 via a competitive mechanism (Ki = 29.4 and 9.1 µM in microsomes and supersomes). Moreover, lurasidone competitively, though weakly diminished the CYP2D6 activity (Ki = 37.5 and 85 µM in microsomes and supersomes).Iloperidone inhibited the activity of CYP3A4 via a noncompetitive mechanism (The examined neuroleptics showed inhibitory effects on different CYP enzymes. The obtained results indicate that metabolic/pharmacokinetic interactions with iloperidone (involving mainly CYP3A4 and CYP2D6) and possibly with lurasidone  may occur during combined therapy. The cytochromes P450 (CYPs) constitute the major family of enzymes capable of catalyzing the oxidative biotransformation of most drugs and other lipophilic xenobiotics, therefore, they are of particular relevance to clinical pharmacology . CYPs ar2A, and dopamine D2 and D3 receptors, moderate affinity binding to dopamine D4, serotonin 5-HT6 and 5-HT7, and norepinephrine α1 receptors, and low-affinity binding to serotonin 5-HT1A, dopamine D1, and histamine H1 receptors. Iloperidone has no appreciable affinity for muscarinic receptors [O-demethylation to generate P89 and undergo carbonyl reduction to metabolite P88 [Iloperidone, a piperidinyl benzisoxazole derivative, is an atypical neuroleptic drug approved for the treatment of acute schizophrenia in adult patients . It prod2, 5-HT2A and 5-HT7 receptors. It also has a high affinity as a partial agonist at the 5-HT1A receptors, and a moderate affinity for adrenergic receptors α2A and α2C [N-dealkylation, hydroxylation of norbornane ring or cyclohexane ring, S-oxidation, reductive cleavage of the isothiazole ring, followed by S-methylation or combinations of two or more of these pathways [Lurasidone, an azapirone derivative with a benzisothiazol-piperazine side chain , is also and α2C , 12. The and α2C . Its majpathways . Lurasidpathways .Among atypical antipsychotic agents, each has a distinctive pharmacokinetic profile including a unique metabolism and excretion pathway. The metabolic pathways engaging particular CYP-enzymes dictate the likelihood that an agent will participate in cytochrome P450-mediated pharmacokinetic drug-drug interactions. Metabolic interactions may proceed via enzyme inhibition or induction, or competition for a specific hepatic CYP enzyme . The potAccording to scientific literature data, possible drug interactions with the atypical neuroleptics iloperidone and lurasidone at a level of cytochrome P450 are not well known. Therefore, the purpose of the present study was to investigate possible inhibitory effects of the above-mentioned drugs on the main CYP enzymes of human liver: CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A4.N-desmethyl caffeine (paraxanthine), diclofenac, 4′-hydroxydiclofenac, bufuralol, 1′-hydroxybufuralol, NADP, NADPH, glucose-6-phosphate, glucose-6-phosphate-dehydrogenase, MgCl2, KCl, ZnSO4, Trizma base and ethylenediaminetetraacetic acid (EDTA) were purchased from Sigma . Testosterone and 6β-testosterone were from Steraloids . All the organic solvents with HPLC purity were supplied by Merck . Pooled human liver microsomes and microsomes from baculovirus-infected insect cells expressing human CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A4 (supersomes) were provided by Corning .Lurasidone and iloperidone were obtained from TargetMol . Caffeine, 3-N-demethylation for CYP1A2 , diclofenac 4′-hydroxylation for CYP2C9 , perazine N-demethylation for CYP2C19 , bufuralol 1′-hydroxylation for CYP2D6 , and testosterone 6β-hydroxylation for CYP3A4 . Incubation systems for CYP2C9, 2C19 and 3A4 contained: 50 mM TRIS/KCL buffer (pH = 7.4), NADPH generating system . Incubation mixture for CYP1A2 included: 0.15 M phosphate buffer (pH = 7.4) and 1 mM NADPH, and for CYP2D6: 0.1 M TRIS/KCL buffer (pH = 7.4), NADPH generating system . The appropriate concentrations of human liver microsomes (0.5 mg/ml for each reaction) or supersomes (50 pmol CYP/ml), various concentrations of a probe substrate in the absence or presence of neuroleptic  were added, The final volume of the reaction mixture was 0.5 ml. The incubation time for supersomes was 30 min (for each reaction) and for liver microsomes: 30 min , 20 min (perazine N-demethylation and testosterone 6β-hydroxylation) or 50 min (caffeine 3-N-demethylation). After the reactions had been stopped, the concentrations of specific substrates and their metabolites formed in liver microsomes or supersomes were assessed by the HPLC method with UV detection (or fluorimetric detection for CYP2D6), as described previously [To study the inhibitory effects of iloperidone and lurasidone on the activity of various CYP isoforms, pooled human liver microsomes and microsomes from baculovirus-infected insect cells expressing human CYPs (supersomes) were used. The following probe reactions were applied, according to the methods previously described –21: caffeviously –21.Km, Vmax, Ki) describing the course of CYP-specific reactions in liver microsomes or supersomes were obtained using the Michaelis–Menten approach and a non-linear regression analysis (Program Sigma Plot 12.3 Enzyme Kinetics). The inhibitory effects of iloperidone and lurasidone on CYP enzymes are presented graphically as Dixon’s plots (1/V against I) indicating Ki values, and Lineweaver–Burk’s plots (1/V against 1/S) showing the mechanism of inhibition .Kinetic parameters  and CYP2D6 . Moreover, iloperidone attenuated the activity of CYP2C19  and CYP1A2 . Iloperidone did not affect the activity of CYP2C9. In comparison, lurasidone moderately inhibited CYP1A2 , CYP2C9 , CYP2C19  and CYP3A4 . Lurasidone weakly diminished the activity of CYP2D6 .To investigate whether iloperidone and lurasidone affect the activity of CYP enzymes, the probe reaction assays were conducted with varied concentration of the neuroleptics. The Dixon`s plots of the metabolism of CYP-specific substrates, carried out in human liver microsomes and supersomes CYP1A2, CYP2D6, CYP2C9, CYP2C19 and CYP3A4, in the absence or presence of the tested neuroleptics, showed that the examined neuroleptics exerted inhibitory effects on different CYP enzymes. However, their potency to inhibit specific CYP enzymes was diverse. Iloperidone exerted a strong inhibitory effect on the activity of CYP3A4 , CYP2C19, CYP2D6 and CYP3A4 activities in pooled human liver microsomes and microsomes from baculovirus-infected insect cells expressing human CYPs (supersomes). However, their potency to inhibit specific CYP enzymes was diverse. Iloperidone potently inhibited CYP3A4 and CYP2D6 via a noncompetitive or competitive mechanism (respectively). Moreover, it moderately diminished the activity of CYP2C19 and weakly affected CYP1A2 via a mixed mechanism. In the case of lurasidone, moderate inhibition of CYP1A2, CYP2C9 and CYP2C19 via a mixed mechanism and of CYP3A4 via a competitive mechanism was observed. The CYP2D6 activity was weakly inhibited via a competitive mechanism.Ki values which are situated in the range of the Ki values observed for such potent CYP3A4 inhibitors as ritonavir, verapamil or erythromycin (Ki = 0.1 − 5 µM) [Ki values calculated for human liver microsomes  are close to the presumed concentration range of iloperidone in the liver of psychiatric patients. Since iloperidone is both a substrate and an inhibitor of CYP3A4 and CYP2D6, it seems possible that the neuroleptic can also inhibit its own metabolism. Iloperidone is also expected to moderately alter the exposure to drugs primarily cleared by CYP2C19 such as diazepam, omeprazole, mephenytoin or tricyclic antidepressants [The potent inhibition of CYP3A4 and CYP2D6 by iloperidone displays  − 5 µM) , as well − 5 µM) , 28. Henressants –32. The ressants .Although lurasidone can be categorized as a moderate inhibitor of CYP2C9, it may have clinically important consequences for some substrates with a narrow therapeutic range . Similarly, the moderate inhibition of CYP1A2 or CYP2C19 by lurasidone demonstrated in vitro in the present study, may be of pharmacological significance in vivo for drugs mainly metabolized or bioactivated by the mentioned enzymes (e.g. bioactivation of antiandrogen flutamide by CYP1A2) .P = 4.43 for the iloperidone and 5.6 for the lurasidone) and related tissue distribution pattern [Albeit the therapeutic plasma concentrations of iloperidone and lurasidone reach the level up to 0.2 and 1.2 μM, respectively , 28, 35, pattern –38. Drug pattern –41, inclDespite the potent inhibition of CYP2C19 and CYP2D6 in human liver microsomes , iloperidone exerts a weaker inhibitory effect on cDNA-expressed CYP2C19 and CYP2D6 in supersomes . A similar situation was observed for lurasidone in the case of the CYP2D6 enzyme, i.e. stronger inhibition occurred in liver microsomes (37.5 µM) than in cDNA-expressed CYP2D6 supersomes (85 µM). This kind of differences in the inhibition potency between human liver microsomes and cDNA expressed CYPs (supersomes) were also observed by Kahma et al. . Zang etThe knowledge of the ability of iloperidone to inhibit CYP3A4, CYP2D6 and possibility of lurasidone to inhibit CYP2C9 is of pharmacological and clinical importance, since these neuroleptics are administered to patients for months or years, and very often in combination with other clinically important drugs that are substrates of the above-mentioned CYP enzymes. Therefore, the inhibition of those CYPs by iloperidone or lurasidone may produce drug-drug interactions. The obtained results suggest that pharmacokinetic interactions involving iloperidone and substrates of CYP2D6  and CYP3A4  or lurasidone and CYP2C9 substrates  are likely to occur in patients during co-administration of the above-mentioned drugs . Given t