International Journal for Parasitology: Drugs and Drug Resistance最新文献

{"title":"Yeast-based assay to identify inhibitors of the malaria parasite sodium phosphate uptake transporter as potential novel antimalarial drugs","authors":"Joseph M. Sweeney ,&nbsp;Ian M. Willis ,&nbsp;Myles H. Akabas","doi":"10.1016/j.ijpddr.2024.100567","DOIUrl":"10.1016/j.ijpddr.2024.100567","url":null,"abstract":"<div><div>Malaria affects almost 250 million people annually and continues to be a significant threat to global public health. Infection with protozoan parasites from the genus <em>Plasmodium</em> causes malaria. The primary treatment for malaria is artemisinin-based combination therapies (ACTs). The spread of ACT-resistant parasites has undermined efforts to control and eradicate malaria. Thus, it is crucial to identify new targets for the development of novel antimalarial drugs. Phosphate is an essential nutrient for all cells. The <em>Plasmodium falciparum</em> genome encodes a single sodium-coupled inorganic phosphate transporter named PfPiT that is essential for parasite proliferation in the asexual blood stage. Thus, PfPiT inhibitors may be promising antimalarial drugs. Like <em>Plasmodium</em>, yeast requires phosphate to grow. We developed a <em>Saccharomyces cerevisiae</em> based growth assay to identify inhibitors of PfPiT. Genome editing was used to create a yeast strain where PfPiT was the only phosphate transporter. Using a radioactive [³²P]phosphate uptake assay, the measured phosphate K_m for PfPiT in yeast was 56 ± 7 μM in 1 mM NaCl at pH 7.4. The K_m decreased to 24 ± 3 μM in 25 mM NaCl consistent with it being a Na⁺ coupled cotransporter. Conditions under which yeast growth was dependent on phosphate uptake mediated by PfPiT were identified and a 22-h growth assay was developed to screen for PfPiT inhibitors. In a screen of 21 compounds, two compounds were identified that inhibited the growth of the PfPiT strain but not that of the parental strain expressing Pho84, one of the five endogenous yeast phosphate transporters. Radioactive phosphate uptake experiments confirmed inhibition of phosphate uptake by the two compounds. The growth inhibition assay provides a simple and inexpensive approach to screen a large compound library for PfPiT inhibitors that may serve as starting points for the development of novel antimalarial drugs.</div></div>","PeriodicalId":13775,"journal":{"name":"International Journal for Parasitology: Drugs and Drug Resistance","volume":"26 ","pages":"Article 100567"},"PeriodicalIF":4.1,"publicationDate":"2024-10-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142499836","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparative proteomic analysis of metronidazole-sensitive and resistant Trichomonas vaginalis suggests a novel mode of metronidazole action and resistance","authors":"Anna-Lena Mayr ,&nbsp;Ana Paunkov ,&nbsp;Karin Hummel ,&nbsp;Ebrahim Razzazi-Fazeli ,&nbsp;David Leitsch","doi":"10.1016/j.ijpddr.2024.100566","DOIUrl":"10.1016/j.ijpddr.2024.100566","url":null,"abstract":"<div><div>The microaerophilic parasite <em>Trichomonas vaginalis</em> occurs worldwide and causes inflammation of the urogenital tract, especially in women. With 156 million infections annually, trichomoniasis is the most prevalent non-viral sexually transmitted disease. Trichomoniasis is treated with 5-nitroimidazoles, especially metronidazole, which are prodrugs that have to be reduced at their nitro group to be activated. Resistance rates to metronidazole have remained comparably low, but they can be higher in certain areas leading to an increase of refractory cases. Metronidazole resistance in <em>T</em>. <em>vaginalis</em> can develop <em>in vivo</em> in clinical isolates, or it can be induced in the laboratory. Both types of resistance share certain characteristics but differ with regard to the dependence of ambient oxygen to become manifest. Although several candidate factors for metronidazole resistance have been described in the past, e.g. pyruvate:ferredoxin oxidoreductase and ferredoxin or thioredoxin reductase, open questions regarding their role in resistance have remained.</div><div>In order to address these questions, we performed a proteomic study with metronidazole-sensitive and –resistant laboratory strains, as well as with clinical strains, in order to identify factors causative for resistance. The list of proteins consistently associated with resistance was surprisingly short. Resistant laboratory and clinical strains only shared the downregulation of flavin reductase 1 (FR1), an enzyme previously identified to be involved in resistance. Originally, FR1 was believed to be an oxygen scavenging enzyme, but here we identified it as a ferric iron reductase which produces ferrous iron. Based on this finding and on further experimental evidence as presented herein, we propose a novel mechanism of metronidazole activation which is based on ferrous iron binding to proteins, thereby rendering them susceptible to complex formation with metronidazole. Upon resolution of iron-protein-metronidazole complexes, metronidazole radicals are formed which quickly react with thiols or proteins in the direct vicinity, leading to breaks in the peptide backbone.</div></div>","PeriodicalId":13775,"journal":{"name":"International Journal for Parasitology: Drugs and Drug Resistance","volume":"26 ","pages":"Article 100566"},"PeriodicalIF":4.1,"publicationDate":"2024-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142377890","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"In artemisinin-resistant falciparum malaria parasites, mitochondrial metabolic pathways are essential for survival but not those of apicoplast","authors":"Manel Ouji ,&nbsp;Thibaud Reyser ,&nbsp;Yoshiki Yamaryo-Botté ,&nbsp;Michel Nguyen ,&nbsp;David Rengel ,&nbsp;Axelle Dutreuil ,&nbsp;Marlène Marcellin ,&nbsp;Odile Burlet-Schiltz ,&nbsp;Jean-Michel Augereau ,&nbsp;Michael K. Riscoe ,&nbsp;Lucie Paloque ,&nbsp;Cyrille Botté ,&nbsp;Françoise Benoit-Vical","doi":"10.1016/j.ijpddr.2024.100565","DOIUrl":"10.1016/j.ijpddr.2024.100565","url":null,"abstract":"<div><div>Emergence and spread of parasite resistance to artemisinins, the first-line antimalarial therapy, threaten the malaria eradication policy. To identify therapeutic targets to eliminate artemisinin-resistant parasites, the functioning of the apicoplast and the mitochondrion was studied, focusing on the fatty acid synthesis type II (FASII) pathway in the apicoplast and the electron transfer chain in the mitochondrion. A significant enrichment of the FASII pathway among the up-regulated genes in artemisinin-resistant parasites under dihydroartemisinin treatment was found, in agreement with published transcriptomic data. However, using GC-MS analyzes of fatty acids, we demonstrated for the first time that the FASII pathway is non-functional, ruling out the use of FASII inhibitors to target artemisinin-resistant parasites. Conversely, by assessing the modulation of the oxygen consumption rate, we evidenced that mitochondrial respiration remains functional and flexible in artemisinin-resistant parasites and even at the quiescent stage. Two novel compounds targeting electron transport chain (ELQ300, ELQ400) efficiently killed quiescent artemisinin-resistant parasites. Therefore, mitochondrial respiration represents a key target for the elimination of artemisinin-resistant persistent <em>Plasmodium falciparum</em> parasites.</div></div>","PeriodicalId":13775,"journal":{"name":"International Journal for Parasitology: Drugs and Drug Resistance","volume":"26 ","pages":"Article 100565"},"PeriodicalIF":4.1,"publicationDate":"2024-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142322497","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Evaluating the amoeba thioredoxin reductase selenoprotein as potential drug target for treatment of Acanthamoeba infections","authors":"Alvie Loufouma-Mbouaka ,&nbsp;Attila Andor ,&nbsp;David Leitsch ,&nbsp;Jorge Pérez-Serrano ,&nbsp;Elias S.J. Arnér ,&nbsp;Julia Walochnik ,&nbsp;Tania Martín-Pérez","doi":"10.1016/j.ijpddr.2024.100564","DOIUrl":"10.1016/j.ijpddr.2024.100564","url":null,"abstract":"<div><div>The genus <em>Acanthamoeba</em> comprises facultative pathogens, causing <em>Acanthamoeba</em> keratitis (AK) and granulomatous amoebic encephalitis (GAE). In both diseases, treatment options are limited, and drug development is challenging. This study aimed to investigate the role of the large thioredoxin reductase selenoprotein of <em>Acanthamoeba</em> (AcTrxR-L) as a potential drug target assessing the effects of the thioredoxin reductase inhibitors auranofin, TRi-1, and TRi-2 on AcTrxR-L activity and on the viability of <em>Acanthamoeba</em> trophozoites. Recombinant expression and purification of AcTrxR-L as a selenoprotein allowed assessments of its enzymatic activity, with reduction of various substrates, including different thioredoxin isoforms. Auranofin demonstrated potent inhibition towards AcTrxR-L, followed by TRi-1, and TRi-2 exhibiting lower effectiveness. The inhibitors showed variable activity against trophozoites in culture, with TRi-1 and TRi-2 resulting in strongly impaired trophozoite viability. Cytotoxicity tests with human corneal epithelial cells revealed lower susceptibility to all compounds compared to <em>Acanthamoeba</em> trophozoites, underscoring their potential as future amoebicidal agents. Altogether, this study highlights the druggability of AcTrxR-L and suggests it to be a promising drug target for the treatment of <em>Acanthamoeba</em> infections. Further research is warranted to elucidate the role of AcTrxR-L in <em>Acanthamoeba</em> pathogenesis and to develop effective therapeutic strategies targeting this redox enzyme.</div></div>","PeriodicalId":13775,"journal":{"name":"International Journal for Parasitology: Drugs and Drug Resistance","volume":"26 ","pages":"Article 100564"},"PeriodicalIF":4.1,"publicationDate":"2024-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142319637","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The phosphatase inhibitor BVT-948 can be used to efficiently screen functional sexual development proteins in the malaria parasite Plasmodium berghei","authors":"Xitong Jia ,&nbsp;Yong Wang ,&nbsp;Meilian Wang ,&nbsp;Hui Min ,&nbsp;Zehou Fang ,&nbsp;Haifeng Lu ,&nbsp;Jiao Li ,&nbsp;Yaming Cao ,&nbsp;Lunhao Bai ,&nbsp;Jinghan Lu","doi":"10.1016/j.ijpddr.2024.100563","DOIUrl":"10.1016/j.ijpddr.2024.100563","url":null,"abstract":"<div><h3>Background</h3><p>Studying and discovering the molecular mechanism of <em>Plasmodium</em> sexual development is crucial for the development of transmission blocking drugs and malaria eradication. The aim of this study was to investigate the feasibility of using phosphatase inhibitors as a tool for screening proteins essential for <em>Plasmodium</em> sexual development and to discover proteins affecting the sexual development of malaria parasites.</p></div><div><h3>Methods</h3><p>Differences in protein phosphorylation among <em>Plasmodium</em> gametocytes incubated with BVT-948 under <em>in vitro</em> ookinete culture conditions were evaluated using phosphoproteomic methods. Gene Ontology (GO) analysis was performed to predict the mechanism by which BVT-948 affected gametocyte–ookinete conversion. The functions of 8 putative proteins involved in <em>Plasmodium berghei</em> sexual development were evaluated. Bioinformatic analysis was used to evaluate the possible mechanism of PBANKA_0100800 in gametogenesis and subsequent sexual development.</p></div><div><h3>Results</h3><p>The phosphorylation levels of 265 proteins decreased while those of 67 increased after treatment with BVT-948. Seven of the 8 genes selected for phenotype screening play roles in <em>P. berghei</em> sexual development, and 4 of these were associated with gametocytogenesis. PBANKA_0100800 plays essential roles in gametocyte–ookinete conversion and transmission to mosquitoes.</p></div><div><h3>Conclusions</h3><p>Seven proteins identified by screening affect <em>P. berghei</em> sexual development, suggesting that phosphatase inhibitors can be used for functional protein screening.</p></div>","PeriodicalId":13775,"journal":{"name":"International Journal for Parasitology: Drugs and Drug Resistance","volume":"26 ","pages":"Article 100563"},"PeriodicalIF":4.1,"publicationDate":"2024-08-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2211320724000447/pdfft?md5=9f49542e884503e417896ae68e574d58&pid=1-s2.0-S2211320724000447-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141995707","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Inhibition of Giardia duodenalis by isocryptolepine -triazole adducts and derivatives","authors":"Supaluk Popruk ,&nbsp;Jumreang Tummatorn ,&nbsp;Suthasinee Sreesai ,&nbsp;Sumate Ampawong ,&nbsp;Tipparat Thiangtrongjit ,&nbsp;Phornpimon Tipthara ,&nbsp;Joel Tarning ,&nbsp;Charnsak Thongsornkleeb ,&nbsp;Somsak Ruchirawat ,&nbsp;Onrapak Reamtong","doi":"10.1016/j.ijpddr.2024.100561","DOIUrl":"10.1016/j.ijpddr.2024.100561","url":null,"abstract":"<div><p><em>Giardia duodenalis</em>, a widespread parasitic flagellate protozoan causing giardiasis, affects millions annually, particularly impacting children and travellers. With no effective vaccine available, treatment primarily relies on the oral administration of drugs targeting trophozoites in the small intestine. However, existing medications pose challenges due to side effects and drug resistance, necessitating the exploration of novel therapeutic options. Isocryptolepine, derived from <em>Cryptolepis sanguinolenta</em>, has demonstrated promising antimicrobial and anticancer properties. This study evaluated eighteen isocryptolepine-triazole adducts for their antigiardial activities and cytotoxicity, with ISO2 demonstrating potent antigiardial activity and minimal cytotoxicity on human intestinal cells. Metabolomics analysis revealed significant alterations in <em>G. duodenalis</em> metabolism upon ISO2 treatment, particularly affecting phospholipid metabolism. Notably, the upregulation of phytosphingosine and triglycerides, and downregulation of certain fatty acids, suggest a profound impact on membrane composition and integrity, potentially contributing to the parasite's demise. Pathway analysis highlighted glycerophospholipid metabolism, cytochrome <em>b</em>5 family heme/steroid binding domain, and P-type ATPase mechanisms as critical pathways affected by ISO2 treatment, underscoring its importance as a potential target for antigiardial therapy. These findings shed light on the mode of action of ISO2 against <em>G. duodenalis</em> and provide valuable insights for further drug development. Moreover, the study also offers a promising avenue for the exploration of isocryptolepine derivatives as novel therapeutic agents for giardiasis, addressing the urgent need for more effective and safer treatment options.</p></div>","PeriodicalId":13775,"journal":{"name":"International Journal for Parasitology: Drugs and Drug Resistance","volume":"26 ","pages":"Article 100561"},"PeriodicalIF":4.1,"publicationDate":"2024-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2211320724000423/pdfft?md5=cf1b4f8e64b6cac6a25597bf8c18ea7c&pid=1-s2.0-S2211320724000423-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141991191","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Benzaldehyde stimulates autophagy via the sonic hedgehog signaling pathway in mouse brain astrocytes after treatment with Angiostrongylus cantonensis excretory-secretory products","authors":"Kuang-Yao Chen ,&nbsp;Chien-Ju Cheng ,&nbsp;Yuan-Ting Chang ,&nbsp;Yi-Hsuan Lin ,&nbsp;Yi-Hao Huang ,&nbsp;Sheng-Yu Lin ,&nbsp;Lian-Chen Wang ,&nbsp;Kai-Yuan Jhan ,&nbsp;Cheng-Hsun Chiu","doi":"10.1016/j.ijpddr.2024.100560","DOIUrl":"10.1016/j.ijpddr.2024.100560","url":null,"abstract":"<div><p>Autophagy is a vital cellular process responsible for digesting various cytoplasmic organelles. This process plays a crucial role in maintaining cell survival and homeostasis, especially under conditions that cause nutrient deficiency, cellular damage, and oxidative stress. Neuroangiostrongyliasis is an infection caused by the parasitic nematode <em>Angiostrongylus cantonensis</em> and is considered as an emerging disease in many parts of the world. However, effective therapeutic strategies for neuroangiostrongyliasis still need to be further developed. In this study, we investigated the effects of benzaldehyde treatment on autophagy and sonic hedgehog (Shh) signaling in <em>A. cantonensis-</em>infected mice and its mechanisms. First, we found autophagosome generation in the central nervous system after <em>A. cantonensis</em> infection. Next, benzaldehyde combined with albendazole treatment reduced eosinophilic meningitis and upregulated the expression of Shh signaling- and autophagy-related molecules in <em>A. cantonensis</em>-infected mouse brains. In vitro experiments demonstrated that benzaldehyde could induce autophagy via the Shh signaling pathway in <em>A. cantonensis</em> excretory-secretory products (ESPs)-treated mouse astrocytes. Finally, benzaldehyde treatment also decreased lipid droplet accumulation and increased cholesterol production by activating the Shh pathway after ESPs treatment. In conclusion, these findings suggested that benzaldehyde treatment could alleviate brain damage by stimulating autophagy generation through the Shh signaling pathway.</p></div>","PeriodicalId":13775,"journal":{"name":"International Journal for Parasitology: Drugs and Drug Resistance","volume":"26 ","pages":"Article 100560"},"PeriodicalIF":4.1,"publicationDate":"2024-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2211320724000411/pdfft?md5=cf24b1d2bd7ceca652003c382746d431&pid=1-s2.0-S2211320724000411-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141985854","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Macrocyclic lactones and ectoparasites control in livestock: Efficacy, drug resistance and therapeutic challenges","authors":"A. Lifschitz ,&nbsp;S. Nava ,&nbsp;V. Miró ,&nbsp;C. Canton ,&nbsp;L. Alvarez ,&nbsp;C. Lanusse","doi":"10.1016/j.ijpddr.2024.100559","DOIUrl":"10.1016/j.ijpddr.2024.100559","url":null,"abstract":"<div><p>Macrocyclic lactones (MLs) are the cornerstone of parasite control in livestock due to their broad-spectrum activity against endo (nematodes) and ecto (lice, ticks, mites) parasites. These molecules, introduced into the veterinary pharmaceutical market 40 years ago, have substantially improved animal welfare and productivity by offering extended high efficacy, reducing treatment frequency, and displaying a favorable safety profile. However, their widespread and intensive use has led to a significant challenge nowadays: <em>the development of parasite resistance</em>. This review focuses on the critical link between drug pharmacokinetics (variation in concentration profiles and exposure over time) and pharmacodynamics (drug efficacy) and the ability of both avermectin and milbemycin MLs families to control livestock ectoparasites. This review discusses the integrated assessment of drug behavior in the host, its diffusion into target parasites, and the impact of different pharmaceutical formulations on enhancing drug delivery to infection sites. These are considered critical research/development areas to optimize the use of MLs, preventing treatment failures and finally extending the lifespan of these essential pharmaceutical ingredients. Finally, the importance of the rational use of MLs, guided by parasite epidemiology and pharmacological knowledge, is emphasized as a key strategy to preserve the antiparasitic efficacy of these still very useful molecules.</p></div>","PeriodicalId":13775,"journal":{"name":"International Journal for Parasitology: Drugs and Drug Resistance","volume":"26 ","pages":"Article 100559"},"PeriodicalIF":4.1,"publicationDate":"2024-08-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S221132072400040X/pdfft?md5=424f13767e5a50ca691d68edb050274c&pid=1-s2.0-S221132072400040X-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141916619","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A multiplexed high throughput screening assay using flow cytometry identifies glycolytic molecular probes in bloodstream form Trypanosoma brucei","authors":"Daniel H. Call ,&nbsp;John Asafo Adjei ,&nbsp;Ryan Pilgrim ,&nbsp;James W. Jeong ,&nbsp;E. Vance Willis ,&nbsp;Ronald A. Zegarra ,&nbsp;Nicholas L. Tapia ,&nbsp;Madalyn Osterhaus ,&nbsp;Jacob A. Vance ,&nbsp;Charles M. Voyton ,&nbsp;James A. Call ,&nbsp;Sabrina S. Pizarro ,&nbsp;James C. Morris ,&nbsp;Kenneth A. Christensen","doi":"10.1016/j.ijpddr.2024.100557","DOIUrl":"10.1016/j.ijpddr.2024.100557","url":null,"abstract":"<div><p>Kinetoplastid organisms, including <em>Trypanosoma brucei</em>, are a significant health burden in many tropical and semitropical countries. Much of their metabolism is poorly understood. To better study kinetoplastid metabolism, chemical probes that inhibit kinetoplastid enzymes are needed. To discover chemical probes, we have developed a high-throughput flow cytometry screening assay that simultaneously measures multiple glycolysis-relevant metabolites in live <em>T. brucei</em> bloodstream form parasites. We transfected parasites with biosensors that measure glucose, ATP, or glycosomal pH. The glucose and ATP sensors were FRET biosensors, while the pH sensor was a GFP-based biosensor. The pH sensor exhibited a different fluorescent profile from the FRET sensors, allowing us to simultaneously measure pH and either glucose or ATP. Cell viability was measured in tandem with the biosensors using thiazole red. We pooled sensor cell lines, loaded them onto plates containing a compound library, and then analyzed them by flow cytometry. The library was analyzed twice, once with the pooled pH and glucose sensor cell lines and once with the pH and ATP sensor cell lines. Multiplexing sensors provided some internal validation of active compounds and gave potential clues for each compound's target(s). We demonstrated this using the glycolytic inhibitor 2-deoxyglucose and the alternative oxidase inhibitor salicylhydroxamic acid. Individual biosensor-based assays exhibited a Z′-factor value acceptable for high-throughput screening, including when multiplexed. We tested assay performance in a pilot screen of 14,976 compounds from the Life Chemicals Compound Library. We obtained hit rates from 0.2 to 0.4% depending on the biosensor, with many compounds impacting multiple sensors. We rescreened 44 hits, and 28 (64%) showed repeatable activity for one or more sensors. One compound exhibited EC₅₀ values in the low micromolar range against two sensors. We expect this method will enable the discovery of glycolytic chemical probes to improve metabolic studies in kinetoplastid parasites.</p></div>","PeriodicalId":13775,"journal":{"name":"International Journal for Parasitology: Drugs and Drug Resistance","volume":"26 ","pages":"Article 100557"},"PeriodicalIF":4.1,"publicationDate":"2024-08-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2211320724000381/pdfft?md5=ee9ca1f5f6eea35ce2b8b29b0d76d4d1&pid=1-s2.0-S2211320724000381-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142008764","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Erratum to “Uncovering the antimalarial potential of toad venoms through a bioassay-guided fractionation process” [Int. J. Parasitol.: Drugs Drug Resist. 20 (2022) 97–107]","authors":"","doi":"10.1016/j.ijpddr.2023.10.002","DOIUrl":"10.1016/j.ijpddr.2023.10.002","url":null,"abstract":"","PeriodicalId":13775,"journal":{"name":"International Journal for Parasitology: Drugs and Drug Resistance","volume":"25 ","pages":"Article 100510"},"PeriodicalIF":4.1,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2211320723000325/pdfft?md5=0d8e8e74e09173394e44da7777759996&pid=1-s2.0-S2211320723000325-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41235103","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}