A Screen against Leishmania Intracellular Amastigotes: Comparison to a Promastigote Screen and Identification of a Host Cell-Specific Hit
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{"title"=>"A screen against leishmania intracellular amastigotes: Comparison to a promastigote screen and identification of a host cell-specific hit", "type"=>"journal", "authors"=>[{"first_name"=>"Geraldine", "last_name"=>"De Muylder", "scopus_author_id"=>"16432905600"}, {"first_name"=>"Kenny K.H.", "last_name"=>"Ang", "scopus_author_id"=>"26535505700"}, {"first_name"=>"Steven", "last_name"=>"Chen", "scopus_author_id"=>"36672657600"}, {"first_name"=>"Michelle R.", "last_name"=>"Arkin", "scopus_author_id"=>"6603802946"}, {"first_name"=>"Juan C.", "last_name"=>"Engel", "scopus_author_id"=>"7402399288"}, {"first_name"=>"James H.", "last_name"=>"McKerrow", "scopus_author_id"=>"35596191600"}], "year"=>2011, "source"=>"PLoS Neglected Tropical Diseases", "identifiers"=>{"pmid"=>"21811648", "doi"=>"10.1371/journal.pntd.0001253", "sgr"=>"79960945898", "isbn"=>"1935-2735 (Electronic) 1935-2727 (Linking)", "scopus"=>"2-s2.0-79960945898", "issn"=>"19352727", "pui"=>"362246868"}, "id"=>"67373fca-cd27-3eaa-9d95-0bfd0042dc91", "abstract"=>"The ability to screen compounds in a high-throughput manner is essential in the process of small molecule drug discovery. Critical to the success of screening strategies is the proper design of the assay, often implying a compromise between ease/speed and a biologically relevant setting. Leishmaniasis is a major neglected disease with limited therapeutic options. In order to streamline efforts for the design of productive drug screens against Leishmania, we compared the efficiency of two screening methods, one targeting the free living and easily cultured promastigote (insect-infective) stage, the other targeting the clinically relevant but more difficult to culture intra-macrophage amastigote (mammal-infective) stage. Screening of a 909-member library of bioactive compounds against Leishmania donovani revealed 59 hits in the promastigote primary screen and 27 in the intracellular amastigote screen, with 26 hits shared by both screens. This suggested that screening against the promastigote stage, although more suitable for automation, fails to identify all active compounds and leads to numerous false positive hits. Of particular interest was the identification of one compound specific to the infective amastigote stage of the parasite. This compound affects intracellular but not axenic parasites, suggesting a host cell-dependent mechanism of action, opening new avenues for anti-leishmanial chemotherapy.", "link"=>"http://www.mendeley.com/research/screen-against-leishmania-intracellular-amastigotes-comparison-promastigote-screen-identification-ho", "reader_count"=>123, "reader_count_by_academic_status"=>{"Unspecified"=>3, "Professor > Associate Professor"=>4, "Librarian"=>1, "Researcher"=>26, "Student > Doctoral Student"=>7, "Student > Ph. D. Student"=>34, "Student > Postgraduate"=>5, "Other"=>5, "Student > Master"=>21, "Student > Bachelor"=>13, "Lecturer > Senior Lecturer"=>1, "Professor"=>3}, "reader_count_by_user_role"=>{"Unspecified"=>3, "Professor > Associate Professor"=>4, "Librarian"=>1, "Researcher"=>26, "Student > Doctoral Student"=>7, "Student > Ph. D. Student"=>34, "Student > Postgraduate"=>5, "Other"=>5, "Student > Master"=>21, "Student > Bachelor"=>13, "Lecturer > Senior Lecturer"=>1, "Professor"=>3}, "reader_count_by_subject_area"=>{"Unspecified"=>8, "Engineering"=>1, "Biochemistry, Genetics and Molecular Biology"=>8, "Mathematics"=>1, "Agricultural and Biological Sciences"=>59, "Medicine and Dentistry"=>11, "Pharmacology, Toxicology and Pharmaceutical Science"=>8, "Veterinary Science and Veterinary Medicine"=>1, "Chemical Engineering"=>1, "Chemistry"=>18, "Social Sciences"=>1, "Immunology and Microbiology"=>6}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>11}, "Chemistry"=>{"Chemistry"=>18}, "Social Sciences"=>{"Social Sciences"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>6}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>59}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>8}, "Mathematics"=>{"Mathematics"=>1}, "Unspecified"=>{"Unspecified"=>8}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>8}, "Chemical Engineering"=>{"Chemical Engineering"=>1}, "Veterinary Science and Veterinary Medicine"=>{"Veterinary Science and Veterinary Medicine"=>1}}, "reader_count_by_country"=>{"United States"=>5, "Brazil"=>3, "United Kingdom"=>2, "Israel"=>1}, "group_count"=>4}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/755495"], "description"=>"<p>White bar: number of compounds identified in both screens. Light grey and black bars: number of compounds specifically active against promastigotes and intracellular amastigotes respectively. Hatched bar: number of compounds active against the promastigote stage but determined as toxic against THP-1 host cell in the intracellular amastigote screen.</p>", "links"=>[], "tags"=>["hits", "intracellular", "amastigote", "promastigote"], "article_id"=>425864, "categories"=>["Microbiology", "Biochemistry", "Infectious Diseases"], "users"=>["Geraldine De Muylder", "Kenny K. H. Ang", "Steven Chen", "Michelle R. Arkin", "Juan C. Engel", "James H. McKerrow"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0001253.g003", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Number_of_hits_identified_with_the_intracellular_amastigote_and_the_promastigote_primary_screens_/425864", "title"=>"Number of hits identified with the intracellular amastigote and the promastigote primary screens.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 16:38:46"}
  • {"files"=>["https://ndownloader.figshare.com/files/755554"], "description"=>"<p>Lower panels: Dose response curve for naloxonazine (left) and naloxone (right) against intracellular amastigotes (black diamonds), promastigotes (black squares), axenic amastigotes (white diamonds) and THP-1 (white triangles) plotting the percentage of parasite growth inhibition.</p>", "links"=>[], "tags"=>["naloxonazine"], "article_id"=>425928, "categories"=>["Microbiology", "Biochemistry", "Infectious Diseases"], "users"=>["Geraldine De Muylder", "Kenny K. H. Ang", "Steven Chen", "Michelle R. Arkin", "Juan C. Engel", "James H. McKerrow"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0001253.g004", "stats"=>{"downloads"=>1, "page_views"=>22, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Structure_and_activity_of_naloxonazine_and_naloxone_/425928", "title"=>"Structure and activity of naloxonazine and naloxone.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 16:39:07"}
  • {"files"=>["https://ndownloader.figshare.com/files/755419"], "description"=>"<p>A. Number of infected THP-1 counted per well treated or not with 1% DMSO or 2 µM amphotericin B. B. Number of parasites counted per well divided by the number of host nuclei per field. C. Dose response curve for amphotericin B plotting the percentage of parasite growth inhibition. Values are mean from at least 3 independent experiments.</p>", "links"=>[], "tags"=>["infected", "thp-1", "dmso", "amphotericin"], "article_id"=>425790, "categories"=>["Microbiology", "Biochemistry", "Infectious Diseases"], "users"=>["Geraldine De Muylder", "Kenny K. H. Ang", "Steven Chen", "Michelle R. Arkin", "Juan C. Engel", "James H. McKerrow"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0001253.g002", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Treatment_of_infected_THP_1_with_DMSO_and_amphotericin_B_/425790", "title"=>"Treatment of infected THP-1 with DMSO and amphotericin B.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 16:38:21"}
  • {"files"=>["https://ndownloader.figshare.com/files/755623"], "description"=>"<p><sup><i>a</i></sup>Selectivity is the ratio between parasite GI<sub>50</sub> and THP-1 GI<sub>50</sub>.</p><p><sup><i>b</i></sup>Specificity is the ratio between promastigote GI<sub>50</sub> and intracellular amastigote GI<sub>50</sub>. Specificity value >2 was the cut-off chosen to define a compound as more active against the intracellular amastigote stage; while a specificity value <0.4 indicated a compound more active against promastigotes; compounds with specificity values between 0.4 and 2 were considered active against both stages.</p>", "links"=>[], "tags"=>["toxicity", "efficacy", "compounds", "intracellular", "promastigotes", "axenic"], "article_id"=>425988, "categories"=>["Microbiology", "Biochemistry", "Infectious Diseases"], "users"=>["Geraldine De Muylder", "Kenny K. H. Ang", "Steven Chen", "Michelle R. Arkin", "Juan C. Engel", "James H. McKerrow"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0001253.t001", "stats"=>{"downloads"=>0, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Host_cell_toxicity_and_efficacy_of_compounds_against_L_donovani_intracellular_amastigotes_promastigotes_and_axenic_amastigotes_/425988", "title"=>"Host cell toxicity and efficacy of compounds against <i>L. donovani</i> intracellular amastigotes, promastigotes and axenic amastigotes.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-20 16:39:25"}
  • {"files"=>["https://ndownloader.figshare.com/files/755313"], "description"=>"<p>A–D. Detection and segmentation of THP-1 host cell and <i>L. donovani</i> intracellular amastigotes. Images obtained with the INCell Analyzer 1000 (20X) of THP-1 cells infected with <i>L. donovani</i> and treated with 1% DMSO (A, B) or 2 µM amphotericin B (C, D). Insert shows the relative fluorescence of DAPI-stained parasite kinetoplast (k) and nucleic DNA (n) and host cell nucleus (N). Segmentation of host cell nuclei and parasite kinetoplast using INCell developer toolbox software (B, D). Red outline: parasite kinetoplast, blue outlines: host cell nucleus and border representing the boundary of the host cell. E. Evolution of the number of parasites and THP-1 host cells in a 72 h time course. THP-1 and <i>L. donovani</i> were counted at several time points after infection using the INCell 1000. White squares: average number of host nuclei per well (n = 8); black circles: average number of parasites counted per well divided by the total number of host nuclei per well (n = 8).</p>", "links"=>[], "tags"=>["thp-1", "segmentation"], "article_id"=>425679, "categories"=>["Microbiology", "Biochemistry", "Infectious Diseases"], "users"=>["Geraldine De Muylder", "Kenny K. H. Ang", "Steven Chen", "Michelle R. Arkin", "Juan C. Engel", "James H. McKerrow"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0001253.g001", "stats"=>{"downloads"=>0, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Infection_of_THP_1_with_L_donovani_detection_segmentation_and_growth_of_host_cell_and_parasite_/425679", "title"=>"Infection of THP-1 with <i>L. donovani</i>: detection, segmentation and growth of host cell and parasite.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 16:37:47"}

PMC Usage Stats | Further Information

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Relative Metric

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