Biophysics of Malarial Parasite Exit from Infected Erythrocytes
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{"title"=>"Biophysics of malarial parasite exit from infected erythrocytes", "type"=>"journal", "authors"=>[{"first_name"=>"Rajesh", "last_name"=>"Chandramohanadas", "scopus_author_id"=>"24832656300"}, {"first_name"=>"YongKeun", "last_name"=>"Park", "scopus_author_id"=>"55494376500"}, {"first_name"=>"Lena", "last_name"=>"Lui", "scopus_author_id"=>"43561402600"}, {"first_name"=>"Ang", "last_name"=>"Li", "scopus_author_id"=>"55648271200"}, {"first_name"=>"David", "last_name"=>"Quinn", "scopus_author_id"=>"16426733700"}, {"first_name"=>"Kingsley", "last_name"=>"Liew", "scopus_author_id"=>"35762295300"}, {"first_name"=>"Monica", "last_name"=>"Diez-Silva", "scopus_author_id"=>"17345537300"}, {"first_name"=>"Yongjin", "last_name"=>"Sung", "scopus_author_id"=>"14326319800"}, {"first_name"=>"Ming", "last_name"=>"Dao", "scopus_author_id"=>"7005492162"}, {"first_name"=>"Chwee Teck", "last_name"=>"Lim", "scopus_author_id"=>"7403654651"}, {"first_name"=>"Peter Rainer", "last_name"=>"Preiser", "scopus_author_id"=>"56346548400"}, {"first_name"=>"Subra", "last_name"=>"Suresh", "scopus_author_id"=>"55433931000"}], "year"=>2011, "source"=>"PLoS ONE", "identifiers"=>{"pui"=>"361966485", "pmid"=>"21698115", "issn"=>"19326203", "doi"=>"10.1371/journal.pone.0020869", "isbn"=>"1932-6203 (Electronic)\\n1932-6203 (Linking)", "scopus"=>"2-s2.0-79959192812", "sgr"=>"79959192812"}, "id"=>"1e47bf2b-9f55-324d-9c73-0eaeb392559d", "abstract"=>"Upon infection and development within human erythrocytes, P. falciparum induces alterations to the infected RBC morphology and bio-mechanical properties to eventually rupture the host cells through parasitic and host derived proteases of cysteine and serine families. We used previously reported broad-spectrum inhibitors (E64d, EGTA-AM and chymostatin) to inhibit these proteases and impede rupture to analyze mechanical signatures associated with parasite escape. Treatment of late-stage iRBCs with E64d and EGTA-AM prevented rupture, resulted in no major RBC cytoskeletal reconfiguration but altered schizont morphology followed by dramatic re-distribution of three-dimensional refractive index (3D-RI) within the iRBC. These phenotypes demonstrated several-fold increased iRBC membrane flickering. In contrast, chymostatin treatment showed no 3D-RI changes and caused elevated fluctuations solely within the parasitophorous vacuole. We show that E64d and EGTA-AM supported PV breakdown and the resulting elevated fluctuations followed non-Gaussian pattern that resulted from direct merozoite impingement against the iRBC membrane. Optical trapping experiments highlighted reduced deformability of the iRBC membranes upon rupture-arrest, more specifically in the treatments that facilitated PV breakdown. Taken together, our experiments provide novel mechanistic interpretations on the role of parasitophorous vacuole in maintaining the spherical schizont morphology, the impact of PV breakdown on iRBC membrane fluctuations leading to eventual parasite escape and the evolution of membrane stiffness properties of host cells in which merozoites were irreversibly trapped, recourse to protease inhibitors. These findings provide a comprehensive, previously unavailable, body of information on the combined effects of biochemical and biophysical factors on parasite egress from iRBCs.", "link"=>"http://www.mendeley.com/research/biophysics-malarial-parasite-exit-infected-erythrocytes", "reader_count"=>65, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>8, "Student > Doctoral Student"=>3, "Researcher"=>14, "Student > Ph. D. Student"=>21, "Student > Postgraduate"=>2, "Other"=>3, "Student > Master"=>5, "Student > Bachelor"=>2, "Lecturer"=>3, "Lecturer > Senior Lecturer"=>1, "Professor"=>3}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>8, "Student > Doctoral Student"=>3, "Researcher"=>14, "Student > Ph. D. 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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/384160", "https://ndownloader.figshare.com/files/384299", "https://ndownloader.figshare.com/files/384444"], "description"=>"<div><p>Upon infection and development within human erythrocytes, <em>P. falciparum</em> induces alterations to the infected RBC morphology and bio-mechanical properties to eventually rupture the host cells through parasitic and host derived proteases of cysteine and serine families. We used previously reported broad-spectrum inhibitors (E64d, EGTA-AM and chymostatin) to inhibit these proteases and impede rupture to analyze mechanical signatures associated with parasite escape. Treatment of late-stage iRBCs with E64d and EGTA-AM prevented rupture, resulted in no major RBC cytoskeletal reconfiguration but altered schizont morphology followed by dramatic re-distribution of three-dimensional refractive index (3D-RI) within the iRBC. These phenotypes demonstrated several-fold increased iRBC membrane flickering. In contrast, chymostatin treatment showed no 3D-RI changes and caused elevated fluctuations solely within the parasitophorous vacuole. We show that E64d and EGTA-AM supported PV breakdown and the resulting elevated fluctuations followed non-Gaussian pattern that resulted from direct merozoite impingement against the iRBC membrane. Optical trapping experiments highlighted reduced deformability of the iRBC membranes upon rupture-arrest, more specifically in the treatments that facilitated PV breakdown. Taken together, our experiments provide novel mechanistic interpretations on the role of parasitophorous vacuole in maintaining the spherical schizont morphology, the impact of PV breakdown on iRBC membrane fluctuations leading to eventual parasite escape and the evolution of membrane stiffness properties of host cells in which merozoites were irreversibly trapped, recourse to protease inhibitors. These findings provide a comprehensive, previously unavailable, body of information on the combined effects of biochemical and biophysical factors on parasite egress from iRBCs.</p> </div>", "links"=>[], "tags"=>["biophysics", "malarial", "parasite", "infected", "erythrocytes"], "article_id"=>135853, "categories"=>["Hematology", "Biochemistry", "Cell Biology", "Microbiology", "Biophysics"], "users"=>["Rajesh Chandramohanadas", "YongKeun Park", "Lena Lui", "Ang Li", "David Quinn", "Kingsley Liew", "Monica Diez-Silva", "Yongjin Sung", "Ming Dao", "Chwee Teck Lim", "Peter Rainer Preiser", "Subra Suresh"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0020869.s001", "https://dx.doi.org/10.1371/journal.pone.0020869.s002", "https://dx.doi.org/10.1371/journal.pone.0020869.s003"], "stats"=>{"downloads"=>11, "page_views"=>17, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Biophysics_of_Malarial_Parasite_Exit_from_Infected_Erythrocytes/135853", "title"=>"Biophysics of Malarial Parasite Exit from Infected Erythrocytes", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2011-06-17 01:37:33"}
  • {"files"=>["https://ndownloader.figshare.com/files/764080"], "description"=>"<p>(<b>A</b>) Cell thickness maps (Upper Panel) and magnitude of membrane fluctuations (Lower Panel) of inhibitor treated iRBCs and corresponding DMSO-treated controls harvested at 46 hpi were generated using diffraction phase microscopy (DPM). (<b>B</b>) Morphology (Upper Panel) and magnitude of membrane fluctuations (Lower panel) from rupture-arrested iRBCs harvested around 50 hpi. Unlike the earlier time point, rupture-phenotypes from E64d and EGTA-treatments showed schizonts that had lost spherical shape and with elevated membrane fluctuations, especially around the edges. Chymostatin-treated, rupture-arrested iRBCs showed no increase in iRBC membrane fluctuations (<b>C</b>) Averaged membrane fluctuations were calculated over the entire area of iRBCs harvested from 46 hpi and 50 hpi. DMSO-treated controls showed increase in fluctuations (accounts for transition from schizont to ring) while E64d and EGTA-AM caused elevated membrane fluctuations in rupture phenotypes. Chymostatin treated samples showed no transition.</p>", "links"=>[], "tags"=>["thickness", "maps", "membrane", "fluctuations", "inhibitor-treated"], "article_id"=>434434, "categories"=>["Hematology", "Biochemistry", "Cell Biology", "Microbiology", "Biophysics"], "users"=>["Rajesh Chandramohanadas", "YongKeun Park", "Lena Lui", "Ang Li", "David Quinn", "Kingsley Liew", "Monica Diez-Silva", "Yongjin Sung", "Ming Dao", "Chwee Teck Lim", "Peter Rainer Preiser", "Subra Suresh"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0020869.g003", "stats"=>{"downloads"=>2, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Cell_thickness_maps_and_membrane_fluctuations_from_inhibitor_treated_iRBCs_/434434", "title"=>"Cell thickness maps and membrane fluctuations from inhibitor-treated iRBCs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-06-17 01:13:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/763965"], "description"=>"<p>Schizont-stage iRBCs (∼44 hpi) treated with DMSO, E64d, EGTA-AM or Chymostatin and harvested at 46 hpi (Upper Panel) and 50–52 hpi (Lower Panel) were analyzed by tomographic phase microscopy (TPM). The images are lateral cross-sections of 3D- refractive index distributions at the focused plane. iRBCs at 46 hpi showed interior structures with low RI, corresponding to parasitophorous vacuole (Upper Panel-white arrow heads). However, inhibitor-treated iRBCs harvested around 52 hpi showed homogeneous distribution of 3D-RI with an apparent lack of visible PV. Scale bar, 5 µm. Color maps show the refractive index (Left) and the Hb concentration (Right). Figure shows representative images from 20 measured tomograms for each group.</p>", "links"=>[], "tags"=>["inhibitors", "treated"], "article_id"=>434318, "categories"=>["Hematology", "Biochemistry", "Cell Biology", "Microbiology", "Biophysics"], "users"=>["Rajesh Chandramohanadas", "YongKeun Park", "Lena Lui", "Ang Li", "David Quinn", "Kingsley Liew", "Monica Diez-Silva", "Yongjin Sung", "Ming Dao", "Chwee Teck Lim", "Peter Rainer Preiser", "Subra Suresh"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0020869.g002", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_3D_RI_distribution_in_inhibitors_treated_iRBCs_/434318", "title"=>"3D-RI distribution in inhibitors treated iRBCs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-06-17 01:11:58"}
  • {"files"=>["https://ndownloader.figshare.com/files/764204"], "description"=>"<p>(<b>A</b>) Shows procedures to analyze the membrane fluctuations of the RBC edge. The coordinates of the edge are retrieved from the measured cell thickness maps. The centre of area is calculated, and then the coordinates of the edge are converted as a function of theta by transforming into polar coordinate system. The instantaneous displacement of edge position, , is calculated by subtracting the time-averaged edge shape, , from the instant edge shape, . (<b>B</b>) Non-Gaussian parameter as a function of mode numbers for the iRBC treated with Chymostation (black line), E64d (red line), and EGTA (blue line), respectively. The error-bars indicate standard errors.</p>", "links"=>[], "tags"=>["membrane", "fluctuations", "rupture-arrested", "irbc", "membranes", "derived", "repeated", "merozoite"], "article_id"=>434556, "categories"=>["Hematology", "Biochemistry", "Cell Biology", "Microbiology", "Biophysics"], "users"=>["Rajesh Chandramohanadas", "YongKeun Park", "Lena Lui", "Ang Li", "David Quinn", "Kingsley Liew", "Monica Diez-Silva", "Yongjin Sung", "Ming Dao", "Chwee Teck Lim", "Peter Rainer Preiser", "Subra Suresh"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0020869.g004", "stats"=>{"downloads"=>3, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Increased_membrane_fluctuations_in_rupture_arrested_iRBC_membranes_are_likely_derived_from_repeated_merozoite_impingement_/434556", "title"=>"Increased membrane fluctuations in rupture-arrested iRBC membranes are likely derived from repeated merozoite impingement.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-06-17 01:15:56"}
  • {"files"=>["https://ndownloader.figshare.com/files/763837"], "description"=>"<p>(<b>A</b>) Schizont-stage iRBCs (∼44 hpi) were treated with DMSO, E64d, EGTA-AM or Chymostatin and were followed by giemsa staining and microscopy. Morphology of iRBCs was analyzed at 46 hpi (Upper Panel) and 55 hpi (Lower Panel). While all schizont-stage iRBCs from DMSO-treated samples ruptured and established ring stage infections by 55<sup>th</sup> hpi, merozoites from inhibitor-treated iRBCs failed to egress. (<b>B</b>) Viability of inhibitor-treated iRBCs was analyzed by JC-1 staining and flow cytometry. The values are expressed as percentage of healthy schizonts gated based on JC-1 and DAPI staining before and after inhibitor treatments; error bars represent standard deviation.</p>", "links"=>[], "tags"=>["inhibitors", "merozoite"], "article_id"=>434200, "categories"=>["Hematology", "Biochemistry", "Cell Biology", "Microbiology", "Biophysics"], "users"=>["Rajesh Chandramohanadas", "YongKeun Park", "Lena Lui", "Ang Li", "David Quinn", "Kingsley Liew", "Monica Diez-Silva", "Yongjin Sung", "Ming Dao", "Chwee Teck Lim", "Peter Rainer Preiser", "Subra Suresh"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0020869.g001", "stats"=>{"downloads"=>4, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Protease_inhibitors_arrest_merozoite_egress_/434200", "title"=>"Protease inhibitors arrest merozoite egress.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-06-17 01:10:00"}
  • {"files"=>["https://ndownloader.figshare.com/files/764307"], "description"=>"<p>Results of Optical tweezers (OT) experiments with iRBCs harvested at 46 hpi and 50 hpi after DMSO, E64d, EGTA-AM and Chymostatin treatments showing shear modulus values <i>(μ)</i> were calculated for each treatment.</p>", "links"=>[], "tags"=>["characterization", "irbc", "membrane"], "article_id"=>434660, "categories"=>["Hematology", "Biochemistry", "Cell Biology", "Microbiology", "Biophysics"], "users"=>["Rajesh Chandramohanadas", "YongKeun Park", "Lena Lui", "Ang Li", "David Quinn", "Kingsley Liew", "Monica Diez-Silva", "Yongjin Sung", "Ming Dao", "Chwee Teck Lim", "Peter Rainer Preiser", "Subra Suresh"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0020869.g005", "stats"=>{"downloads"=>6, "page_views"=>15, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Optical_trapping_characterization_of_iRBC_membrane_rigidity_/434660", "title"=>"Optical trapping characterization of iRBC membrane rigidity.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-06-17 01:17:40"}

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Relative Metric

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