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      Robust continuous in vitro culture of the Plasmodium cynomolgi erythrocytic stages

      research-article
      1 , 2 , 3 , 2 , 3 , 1 , 4 , 1 , 2 , 4 , 5 , 6 , 7 , 4 , 4 , 3 , 3 , 3 , 7 , 8 , 9 , 4 , 10 , 10 , 10 , 7 , 11 , 12 , 3 , 3 , 13 , 14 , 9 , 15 , 16 , 16 , 6 , 1 , 4 , 7 , 3 , 13 , 14 , 16 , 2 , , 1 , 3 , 4 , 17 ,
      Nature Communications
      Nature Publishing Group UK
      Malaria, Model protists, High-throughput screening

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          Abstract

          The ability to culture pathogenic organisms substantially enhances the quest for fundamental knowledge and the development of vaccines and drugs. Thus, the elaboration of a protocol for the in vitro cultivation of the erythrocytic stages of Plasmodium falciparum revolutionized research on this important parasite. However, for P. vivax, the most widely distributed and difficult to treat malaria parasite, a strict preference for reticulocytes thwarts efforts to maintain it in vitro. Cultivation of P. cynomolgi, a macaque-infecting species phylogenetically close to P. vivax, was briefly reported in the early 1980s, but not pursued further. Here, we define the conditions under which P. cynomolgi can be adapted to long term in vitro culture to yield parasites that share many of the morphological and phenotypic features of P. vivax. We further validate the potential of this culture system for high-throughput screening to prime and accelerate anti- P. vivax drug discovery efforts.

          Abstract

          Present understanding of Plasmodium vivax biology is hampered by its inability to grow in vitro. Here, the authors developed an in vitro culture of its simian counterpart, P. cynomolgi, which shares morphological and phenotypic similarities with P. vivax, initiating a new phase in vivax research.

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          Most cited references60

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          A Simple Statistical Parameter for Use in Evaluation and Validation of High Throughput Screening Assays

          J-H Zhang (1999)
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            Spiroindolones, a potent compound class for the treatment of malaria.

            Recent reports of increased tolerance to artemisinin derivatives--the most recently adopted class of antimalarials--have prompted a need for new treatments. The spirotetrahydro-beta-carbolines, or spiroindolones, are potent drugs that kill the blood stages of Plasmodium falciparum and Plasmodium vivax clinical isolates at low nanomolar concentration. Spiroindolones rapidly inhibit protein synthesis in P. falciparum, an effect that is ablated in parasites bearing nonsynonymous mutations in the gene encoding the P-type cation-transporter ATPase4 (PfATP4). The optimized spiroindolone NITD609 shows pharmacokinetic properties compatible with once-daily oral dosing and has single-dose efficacy in a rodent malaria model.
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              Microscale culture of human liver cells for drug development.

              Tissue function depends on hierarchical structures extending from single cells ( approximately 10 microm) to functional subunits (100 microm-1 mm) that coordinate organ functions. Conventional cell culture disperses tissues into single cells while neglecting higher-order processes. The application of semiconductor-driven microtechnology in the biomedical arena now allows fabrication of microscale tissue subunits that may be functionally improved and have the advantages of miniaturization. Here we present a miniaturized, multiwell culture system for human liver cells with optimized microscale architecture that maintains phenotypic functions for several weeks. The need for such models is underscored by the high rate of pre-launch and post-market attrition of pharmaceuticals due to liver toxicity. We demonstrate utility through assessment of gene expression profiles, phase I/II metabolism, canalicular transport, secretion of liver-specific products and susceptibility to hepatotoxins. The combination of microtechnology and tissue engineering may enable development of integrated tissue models in the so-called 'human on a chip'.
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                Author and article information

                Contributors
                b.russell@otago.ac.nz
                micpb@nus.edu.sg
                Journal
                Nat Commun
                Nat Commun
                Nature Communications
                Nature Publishing Group UK (London )
                2041-1723
                12 August 2019
                12 August 2019
                2019
                : 10
                : 3635
                Affiliations
                [1 ]ISNI 0000 0004 0387 2429, GRID grid.430276.4, Singapore Immunology Network, A*STAR, ; Singapore, 138648 Singapore
                [2 ]ISNI 0000 0004 1936 7830, GRID grid.29980.3a, Department of Microbiology and Immunology, , University of Otago, ; Dunedin, 9054 New Zealand
                [3 ]GRID grid.410761.5, Novartis Institute for Tropical Diseases, ; Singapore, 138670 Singapore
                [4 ]ISNI 0000 0001 2180 6431, GRID grid.4280.e, Department of Microbiology and Immunology, Yong Loo Lin School of Medicine, , National University of Singapore, ; Singapore, 119077 Singapore
                [5 ]ISNI 0000 0004 1937 0490, GRID grid.10223.32, Department of Parasitology and Entomology, Faculty of Public Health, , Mahidol University, ; Bangkok, 10400 Thailand
                [6 ]ISNI 0000 0004 0625 2495, GRID grid.11184.3d, Department of Parasitology, , Biomedical Primate Research Centre, ; Rijswijk, 2288 The Netherlands
                [7 ]ISNI 0000 0004 1936 738X, GRID grid.213876.9, Center for Tropical and Emerging Global Diseases, , University of Georgia, ; Athens, 30602 USA
                [8 ]ISNI 0000 0001 0941 6502, GRID grid.189967.8, Division of Pulmonary, Allergy, Critical Care & Sleep Medicine, , Emory University, ; Atlanta, 30322 USA
                [9 ]ISNI 0000 0001 0941 6502, GRID grid.189967.8, Emory Vaccine Center, , Emory University, ; Atlanta, 30317 USA
                [10 ]ISNI 0000 0004 0439 2056, GRID grid.418424.f, Laboratory Animal Services, Scientific Operations, , Novartis Institutes for Biomedical Research, ; East Hanover, 07936-1080 USA
                [11 ]ISNI 0000 0004 1937 0490, GRID grid.10223.32, Shoklo Malaria Research Unit, Mahidol-Oxford Tropical Medicine Research Unit, Faculty of Tropical Medicine, , Mahidol University, ; Mae Sot, 63110 Thailand
                [12 ]ISNI 0000 0004 1936 8948, GRID grid.4991.5, Centre for Tropical Medicine and Global Health, Nuffield Department of Medicine Research Building, , University of Oxford Old Road Campus, ; Oxford, OX3 7FZ UK
                [13 ]GRID grid.463810.8, Sorbonne Universités, UPMC Univ Paris 06, CR7, Centre d’Immunologie et des Maladies Infectieuses (CIMI-Paris), ; Paris, F-75013 France
                [14 ]ISNI 0000 0001 2112 9282, GRID grid.4444.0, CIMI-Paris, INSERM, U1135, , CNRS, ; Paris, F-75013 France
                [15 ]ISNI 0000 0001 0941 6502, GRID grid.189967.8, Division of Infectious Diseases, Department of Medicine, , Emory University, ; Atlanta, 30322 USA
                [16 ]ISNI 0000 0001 2171 2558, GRID grid.5842.b, CEA-Université Paris Sud 11-INSERM U1184, Immunology of Viral Infections and Autoimmune Diseases (IMVA), IDMIT Department, ; IBJF, DRF, Fontenay-aux-Roses, 92265 France
                [17 ]ISNI 0000 0004 0425 469X, GRID grid.8991.9, Faculty of Infectious and Tropical Diseases, , London School of Hygiene & Tropical Medicine, ; London, WC1E 7HT UK
                Author information
                http://orcid.org/0000-0001-9658-7528
                http://orcid.org/0000-0001-9022-729X
                http://orcid.org/0000-0002-9318-9310
                http://orcid.org/0000-0002-9560-5656
                http://orcid.org/0000-0002-7951-0745
                http://orcid.org/0000-0003-0349-1557
                http://orcid.org/0000-0002-0238-965X
                http://orcid.org/0000-0003-2333-4348
                http://orcid.org/0000-0001-9651-6439
                Article
                11332
                10.1038/s41467-019-11332-4
                6690977
                30602773
                de5679aa-38bb-496a-8df2-70afd0da2087
                © The Author(s) 2019

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 22 October 2018
                : 28 June 2019
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                © The Author(s) 2019

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                malaria,model protists,high-throughput screening
                Uncategorized
                malaria, model protists, high-throughput screening

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