P. falciparum In Vitro Killing Rates Allow to Discriminate between Different Antimalarial Mode-of-Action
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{"title"=>"P. falciparum in vitro killing rates allow to discriminate between different antimalarial mode-of-action", "type"=>"journal", "authors"=>[{"first_name"=>"Laura M.", "last_name"=>"Sanz", "scopus_author_id"=>"56912989800"}, {"first_name"=>"Benigno", "last_name"=>"Crespo", "scopus_author_id"=>"54419808000"}, {"first_name"=>"Cristina", "last_name"=>"De-Cózar", "scopus_author_id"=>"36080303600"}, {"first_name"=>"Xavier C.", "last_name"=>"Ding", "scopus_author_id"=>"55038591000"}, {"first_name"=>"Jose L.", "last_name"=>"Llergo", "scopus_author_id"=>"55039446600"}, {"first_name"=>"Jeremy N.", "last_name"=>"Burrows", "scopus_author_id"=>"55352481100"}, {"first_name"=>"Jose F.", "last_name"=>"García-Bustos", "scopus_author_id"=>"6701821840"}, {"first_name"=>"Francisco Javier", "last_name"=>"Gamo", "scopus_author_id"=>"6603169959"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "scopus"=>"2-s2.0-84857591058", "pui"=>"364337926", "doi"=>"10.1371/journal.pone.0030949", "isbn"=>"1932-6203 (Electronic) 1932-6203 (Linking)", "sgr"=>"84857591058", "pmid"=>"22383983"}, "id"=>"cb6fd5eb-d30e-33d7-8b6e-8e6f45b48b4b", "abstract"=>"Chemotherapy is still the cornerstone for malaria control. Developing drugs against Plasmodium parasites and monitoring their efficacy requires methods to accurately determine the parasite killing rate in response to treatment. Commonly used techniques essentially measure metabolic activity as a proxy for parasite viability. However, these approaches are susceptible to artefacts, as viability and metabolism are two parameters that are coupled during the parasite life cycle but can be differentially affected in response to drug actions. Moreover, traditional techniques do not allow to measure the speed-of-action of compounds on parasite viability, which is an essential efficacy determinant. We present here a comprehensive methodology to measure in vitro the direct effect of antimalarial compounds over the parasite viability, which is based on limiting serial dilution of treated parasites and re-growth monitoring. This methodology allows to precisely determine the killing rate of antimalarial compounds, which can be quantified by the parasite reduction ratio and parasite clearance time, which are key mode-of-action parameters. Importantly, we demonstrate that this technique readily permits to determine compound killing activities that might be otherwise missed by traditional, metabolism-based techniques. The analysis of a large set of antimalarial drugs reveals that this viability-based assay allows to discriminate compounds based on their antimalarial mode-of-action. This approach has been adapted to perform medium throughput screening, facilitating the identification of fast-acting antimalarial compounds, which are crucially needed for the control and possibly the eradication of malaria.", "link"=>"http://www.mendeley.com/research/p-falciparum-vitro-killing-rates-allow-discriminate-between-different-antimalarial-modeofaction", "reader_count"=>109, "reader_count_by_academic_status"=>{"Unspecified"=>4, "Professor > Associate Professor"=>1, "Researcher"=>33, "Student > Doctoral Student"=>5, "Student > Ph. D. Student"=>26, "Student > Postgraduate"=>4, "Student > Master"=>13, "Other"=>14, "Student > Bachelor"=>4, "Lecturer"=>2, "Professor"=>3}, "reader_count_by_user_role"=>{"Unspecified"=>4, "Professor > Associate Professor"=>1, "Researcher"=>33, "Student > Doctoral Student"=>5, "Student > Ph. D. Student"=>26, "Student > Postgraduate"=>4, "Student > Master"=>13, "Other"=>14, "Student > Bachelor"=>4, "Lecturer"=>2, "Professor"=>3}, "reader_count_by_subject_area"=>{"Unspecified"=>8, "Biochemistry, Genetics and Molecular Biology"=>11, "Mathematics"=>1, "Agricultural and Biological Sciences"=>46, "Medicine and Dentistry"=>10, "Pharmacology, Toxicology and Pharmaceutical Science"=>6, "Business, Management and Accounting"=>1, "Chemistry"=>23, "Social Sciences"=>1, "Immunology and Microbiology"=>2}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>10}, "Chemistry"=>{"Chemistry"=>23}, "Social Sciences"=>{"Social Sciences"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>2}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>46}, "Business, Management and Accounting"=>{"Business, Management and Accounting"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>11}, "Mathematics"=>{"Mathematics"=>1}, "Unspecified"=>{"Unspecified"=>8}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>6}}, "reader_count_by_country"=>{"Japan"=>1, "Brazil"=>1, "South Africa"=>2, "United Kingdom"=>1, "France"=>1, "Portugal"=>1}, "group_count"=>6}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/676620"], "description"=>"a<p>estimation based on the first 24 hours of treatment.</p>", "links"=>[], "tags"=>["parasite", "clearance", "antimalarial"], "article_id"=>347105, "categories"=>["Microbiology", "Biochemistry", "Infectious Diseases", "Pharmacology"], "users"=>["Laura M. Sanz", "Benigno Crespo", "Cristina De-Cózar", "Xavier C. Ding", "Jose L. Llergo", "Jeremy N. Burrows", "Jose F. García-Bustos", "Francisco-Javier Gamo"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0030949.t001", "stats"=>{"downloads"=>8, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_In_vitro_parasite_reduction_ratio_and_clearance_time_in_response_to_classical_antimalarial_drugs_/347105", "title"=>"<i>In vitro</i> parasite reduction ratio and clearance time in response to classical antimalarial drugs.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2012-02-23 01:58:25"}
  • {"files"=>["https://ndownloader.figshare.com/files/676105"], "description"=>"<p><b>A</b>. Intraerythrocytic <i>P. falciparum</i> cultured at 0.5% parasitemia and 2% hematocrit is treated with drugs. The medium is exchanged and the drug replenished every 24 hours. Aliquots corresponding to 10<sup>5</sup> parasites are taken out at defined time points, washed, and free-drug parasites cultured with fresh erythrocytes under limiting serial dilution conditions (see Material and Methods). Parasite growth is subsequently monitored after 21 days and confirmed after 28 days, allowing to calculate the initial number of viable parasite in the aliquot. <b>B</b>. Parasite viability measurement allows in turn to determine the drug lag phase (i.e. time needed to reach the maximal rate of killing), PRR over one life cycle, and 99.9% PCT (i.e. the time needed to decrease the number of viable parasites by 3 –log units). The data presented in this panel are for illustration purpose only. Axe Y shows log (viable parasites +1) to allow representation of logarithms when counting of number of viable parasites is equal to zero.</p>", "links"=>[], "tags"=>["prr"], "article_id"=>346596, "categories"=>["Microbiology", "Biochemistry", "Infectious Diseases", "Pharmacology"], "users"=>["Laura M. Sanz", "Benigno Crespo", "Cristina De-Cózar", "Xavier C. Ding", "Jose L. Llergo", "Jeremy N. Burrows", "Jose F. García-Bustos", "Francisco-Javier Gamo"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0030949.g001", "stats"=>{"downloads"=>0, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Schematic_representation_of_the_in_vitro_PRR_assay_/346596", "title"=>"Schematic representation of the <i>in vitro</i> PRR assay.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-23 01:49:56"}
  • {"files"=>["https://ndownloader.figshare.com/files/676239"], "description"=>"<p><b>A. </b><i>P. falciparum</i> viability time-course profiles for chloroquine (chq), mefloquine (mef), piperaquine (pip), artemisinin (art), lumefantrine (lum), pyronaridine (pyro), pyrimethamine (pyri), and atovaquone (ato). Error bars represent the SEM of at least 4 independent experiments. <b>B</b>. and <b>C</b>. Scatter plots of the compounds tested reporting the IC<sub>50</sub> versus the log(PRR) and 99.9% PCT, respectively. The dotted line in panel B is a log linear regression, the slope thereof is not significantly different from zero (<i>p</i> = 0.48). Data of panel C do not converge enough to establish a regression line.</p>", "links"=>[], "tags"=>["viability", "antimalarial"], "article_id"=>346728, "categories"=>["Microbiology", "Biochemistry", "Infectious Diseases", "Pharmacology"], "users"=>["Laura M. Sanz", "Benigno Crespo", "Cristina De-Cózar", "Xavier C. Ding", "Jose L. Llergo", "Jeremy N. Burrows", "Jose F. García-Bustos", "Francisco-Javier Gamo"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0030949.g003", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Parasite_viability_in_response_to_various_classical_antimalarial_drugs_/346728", "title"=>"Parasite viability in response to various classical antimalarial drugs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-23 01:52:08"}
  • {"files"=>["https://ndownloader.figshare.com/files/676310"], "description"=>"<p><b>A</b>. <i>P. falciparum</i> viability time-course profiles for atovaquone, pyrimethamine, and artemisinin at concentrations corresponding to 1×, 3×, 10×, and 100× their respective IC<sub>50</sub>. Error bars are SEM of at least 4 independent experiments. <b>B</b>. Values represented in panel A. No PRR or 99.9% PCT could be calculated for the 1× IC<sub>50</sub> conditions.</p>", "links"=>[], "tags"=>["viability", "time-course", "concentrations"], "article_id"=>346794, "categories"=>["Microbiology", "Biochemistry", "Infectious Diseases", "Pharmacology"], "users"=>["Laura M. Sanz", "Benigno Crespo", "Cristina De-Cózar", "Xavier C. Ding", "Jose L. Llergo", "Jeremy N. Burrows", "Jose F. García-Bustos", "Francisco-Javier Gamo"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0030949.g004", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Parasite_viability_time_course_in_response_to_various_concentrations_of_drugs_/346794", "title"=>"Parasite viability time-course in response to various concentrations of drugs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-23 01:53:14"}
  • {"files"=>["https://ndownloader.figshare.com/files/676172"], "description"=>"<p><b>A</b>. <i>P. falciparum</i> radio-labeled hypoxanthine incorporations after 48 hours of drug treatment at concentrations corresponding to 10× IC<sub>50</sub> with artemisinin, atovaquone, and azithromycin, reported as percentages of untreated controls. Data are averages of 12 repetitions from two independent experiments <b>B</b>. <i>P. falciparum</i> viability after 48 hours of drug treatment at concentrations corresponding to 10× IC<sub>50</sub> with artemisinin, atovaquone, and azithromycin, reported as number of viable parasite, as determined by limiting serial dilutions. Data are averages of 4 independent experiments. In both panels, error bars represent the standard error of the mean (SEM).</p>", "links"=>[], "tags"=>["metabolic", "viability"], "article_id"=>346660, "categories"=>["Microbiology", "Biochemistry", "Infectious Diseases", "Pharmacology"], "users"=>["Laura M. Sanz", "Benigno Crespo", "Cristina De-Cózar", "Xavier C. Ding", "Jose L. Llergo", "Jeremy N. Burrows", "Jose F. García-Bustos", "Francisco-Javier Gamo"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0030949.g002", "stats"=>{"downloads"=>0, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Comparison_of_metabolic_and_viability_assays_/346660", "title"=>"Comparison of metabolic and viability assays.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-23 01:51:00"}
  • {"files"=>["https://ndownloader.figshare.com/files/676380"], "description"=>"<p><i>P. falciparum</i> viability time-course profiles of four classical antimalarial drugs, illustrating the wide range of speed-of-action measured. Viable parasite are cleared in response to fast-acting (art and chq), but not slow-acting (pyri and ato) drugs after 72 hours of treatment.</p>", "links"=>[], "tags"=>["antimalarial"], "article_id"=>346869, "categories"=>["Microbiology", "Biochemistry", "Infectious Diseases", "Pharmacology"], "users"=>["Laura M. Sanz", "Benigno Crespo", "Cristina De-Cózar", "Xavier C. Ding", "Jose L. Llergo", "Jeremy N. Burrows", "Jose F. García-Bustos", "Francisco-Javier Gamo"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0030949.g005", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Classical_antimalarial_killing_rate_profiles_/346869", "title"=>"Classical antimalarial killing rate profiles.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-23 01:54:29"}
  • {"files"=>["https://ndownloader.figshare.com/files/345631", "https://ndownloader.figshare.com/files/345669", "https://ndownloader.figshare.com/files/345709", "https://ndownloader.figshare.com/files/345756", "https://ndownloader.figshare.com/files/345803", "https://ndownloader.figshare.com/files/345850", "https://ndownloader.figshare.com/files/345918", "https://ndownloader.figshare.com/files/345976", "https://ndownloader.figshare.com/files/346043"], "description"=>"<div><p>Chemotherapy is still the cornerstone for malaria control. Developing drugs against <em>Plasmodium</em> parasites and monitoring their efficacy requires methods to accurately determine the parasite killing rate in response to treatment. Commonly used techniques essentially measure metabolic activity as a proxy for parasite viability. However, these approaches are susceptible to artefacts, as viability and metabolism are two parameters that are coupled during the parasite life cycle but can be differentially affected in response to drug actions. Moreover, traditional techniques do not allow to measure the speed-of-action of compounds on parasite viability, which is an essential efficacy determinant. We present here a comprehensive methodology to measure <em>in vitro</em> the direct effect of antimalarial compounds over the parasite viability, which is based on limiting serial dilution of treated parasites and re-growth monitoring. This methodology allows to precisely determine the killing rate of antimalarial compounds, which can be quantified by the parasite reduction ratio and parasite clearance time, which are key mode-of-action parameters. Importantly, we demonstrate that this technique readily permits to determine compound killing activities that might be otherwise missed by traditional, metabolism-based techniques. The analysis of a large set of antimalarial drugs reveals that this viability-based assay allows to discriminate compounds based on their antimalarial mode-of-action. This approach has been adapted to perform medium throughput screening, facilitating the identification of fast-acting antimalarial compounds, which are crucially needed for the control and possibly the eradication of malaria.</p> </div>", "links"=>[], "tags"=>["falciparum", "rates", "discriminate", "antimalarial", "mode-of-action"], "article_id"=>128324, "categories"=>["Cancer", "Biochemistry", "Microbiology", "Pharmacology"], "users"=>["Laura M. Sanz", "Benigno Crespo", "Cristina De-Cózar", "Xavier C. Ding", "Jose L. Llergo", "Jeremy N. Burrows", "Jose F. García-Bustos", "Francisco-Javier Gamo"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0030949.s001", "https://dx.doi.org/10.1371/journal.pone.0030949.s002", "https://dx.doi.org/10.1371/journal.pone.0030949.s003", "https://dx.doi.org/10.1371/journal.pone.0030949.s004", "https://dx.doi.org/10.1371/journal.pone.0030949.s005", "https://dx.doi.org/10.1371/journal.pone.0030949.s006", "https://dx.doi.org/10.1371/journal.pone.0030949.s007", "https://dx.doi.org/10.1371/journal.pone.0030949.s008", "https://dx.doi.org/10.1371/journal.pone.0030949.s009"], "stats"=>{"downloads"=>16, "page_views"=>43, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_P_falciparum_In_Vitro_Killing_Rates_Allow_to_Discriminate_between_Different_Antimalarial_Mode_of_Action/128324", "title"=>"<em>P. falciparum In Vitro</em> Killing Rates Allow to Discriminate between Different Antimalarial Mode-of-Action", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2012-02-23 02:18:44"}
  • {"files"=>["https://ndownloader.figshare.com/files/676552"], "description"=>"<p><i>P. falciparum</i> viability after 72 hours of treatment with the indicated drugs, reported as the log of viable parasite+1, as compared to the untreated controls. The red line represents the threshold of 99.9% parasite reduction. The data are representative of at least 3 independent experiments.</p>", "links"=>[], "tags"=>["time-point", "viability"], "article_id"=>347038, "categories"=>["Microbiology", "Biochemistry", "Infectious Diseases", "Pharmacology"], "users"=>["Laura M. Sanz", "Benigno Crespo", "Cristina De-Cózar", "Xavier C. Ding", "Jose L. Llergo", "Jeremy N. Burrows", "Jose F. García-Bustos", "Francisco-Javier Gamo"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0030949.g007", "stats"=>{"downloads"=>1, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Single_time_point_viability_measurement_/347038", "title"=>"Single time-point viability measurement.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-23 01:57:18"}
  • {"files"=>["https://ndownloader.figshare.com/files/676476"], "description"=>"<p><i>P. falciparum</i> viability time-course profiles for artemisinin, atovaquone, GW648495, and GW844520 measured at 0, 24, 48, 72, 96, and 120 hours. Artemether and artesunate have been investigated at 24 hours only. Error bars are SEM of at least 4 independent experiments.</p>", "links"=>[], "tags"=>["viability", "correlates", "mode-of-action"], "article_id"=>346965, "categories"=>["Microbiology", "Biochemistry", "Infectious Diseases", "Pharmacology"], "users"=>["Laura M. Sanz", "Benigno Crespo", "Cristina De-Cózar", "Xavier C. Ding", "Jose L. Llergo", "Jeremy N. Burrows", "Jose F. García-Bustos", "Francisco-Javier Gamo"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0030949.g006", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Parasite_viability_correlates_with_the_mode_of_action_of_antimalarials_/346965", "title"=>"Parasite viability correlates with the mode-of-action of antimalarials.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-23 01:56:05"}

PMC Usage Stats | Further Information

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