Mechanisms of Fatal Cardiotoxicity following High-Dose Cyclophosphamide Therapy and a Method for Its Prevention
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{"title"=>"Mechanisms of fatal cardiotoxicity following high-dose cyclophosphamide therapy and a method for its prevention", "type"=>"journal", "authors"=>[{"first_name"=>"Takuro", "last_name"=>"Nishikawa", "scopus_author_id"=>"14123740800"}, {"first_name"=>"Emiko", "last_name"=>"Miyahara", "scopus_author_id"=>"44761539400"}, {"first_name"=>"Koichiro", "last_name"=>"Kurauchi", "scopus_author_id"=>"55608812700"}, {"first_name"=>"Erika", "last_name"=>"Watanabe", "scopus_author_id"=>"55227191400"}, {"first_name"=>"Kazuro", "last_name"=>"Ikawa", "scopus_author_id"=>"11540070100"}, {"first_name"=>"Kousuke", "last_name"=>"Asaba", "scopus_author_id"=>"56747640600"}, {"first_name"=>"Takayuki", "last_name"=>"Tanabe", "scopus_author_id"=>"35312484900"}, {"first_name"=>"Yasuhiro", "last_name"=>"Okamoto", "scopus_author_id"=>"35311895200"}, {"first_name"=>"Yoshifumi", "last_name"=>"Kawano", "scopus_author_id"=>"7201460654"}], "year"=>2015, "source"=>"PLoS ONE", "identifiers"=>{"sgr"=>"84938377831", "doi"=>"10.1371/journal.pone.0131394", "issn"=>"19326203", "pui"=>"605407902", "pmid"=>"26114497", "scopus"=>"2-s2.0-84938377831"}, "id"=>"44a19f51-f48b-33e5-a7a0-f38b201a49b6", "abstract"=>"Observed only after administration of high doses, cardiotoxicity is the dose-limiting effect of cyclophosphamide (CY). We investigated the poorly understood cardiotoxic mechanisms of high-dose CY. A rat cardiac myocardial cell line, H9c2, was exposed to CY metabolized by S9 fraction of rat liver homogenate mixed with co-factors (CYS9). Cytotoxicity was then evaluated by 3-(4,5-dimethyl-2-thiazolyl)¬2,5-diphenyl¬2H-tetrazolium bromide (MTT) assay, lactate dehydrogenase release, production of reactive oxygen species (ROS), and incidence of apoptosis. We also investigated how the myocardial cellular effects of CYS9 were modified by acrolein scavenger N-acetylcysteine (NAC), antioxidant isorhamnetin (ISO), and CYP inhibitor β-ionone (BIO). Quantifying CY and CY metabolites by means of liquid chromatography coupled with electrospray tandem mass spectrometry, we assayed culture supernatants of CYS9 with and without candidate cardioprotectant agents. Assay results for MTT showed that treatment with CY (125-500 μM) did not induce cytotoxicity. CYS9, however, exhibited myocardial cytotoxicity when CY concentration was 250 μM or more. After 250 μM of CY was metabolized in S9 mix for 2 h, the concentration of CY was 73.6 ± 8.0 μM, 4-hydroxy-cyclophosphamide (HCY) 17.6 ± 4.3, o-carboxyethyl-phosphoramide (CEPM) 26.6 ± 5.3 μM, and acrolein 26.7 ± 2.5 μM. Inhibition of CYS9-induced cytotoxicity occurred with NAC, ISO, and BIO. When treated with ISO or BIO, metabolism of CY was significantly inhibited. Pre-treatment with NAC, however, did not inhibit the metabolism of CY: compared to control samples, we observed no difference in HCY, a significant increase of CEPM, and a significant decrease of acrolein. Furthermore, NAC pre-treatment did not affect intracellular amounts of ROS produced by CYS9. Since acrolein seems to be heavily implicated in the onset of cardiotoxicity, any competitive metabolic processing of CY that reduces its transformation to acrolein is likely to be an important mechanism for preventing cardiotoxicity.", "link"=>"http://www.mendeley.com/research/mechanisms-fatal-cardiotoxicity-following-highdose-cyclophosphamide-therapy-method-prevention", "reader_count"=>19, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Researcher"=>4, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>5, "Student > Postgraduate"=>3, "Student > Master"=>1, "Student > Bachelor"=>4}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Researcher"=>4, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>5, "Student > Postgraduate"=>3, "Student > Master"=>1, "Student > Bachelor"=>4}, "reader_count_by_subject_area"=>{"Unspecified"=>2, "Biochemistry, Genetics and Molecular Biology"=>2, "Medicine and Dentistry"=>9, "Agricultural and Biological Sciences"=>1, "Pharmacology, Toxicology and Pharmaceutical Science"=>2, "Psychology"=>1, "Computer Science"=>1, "Immunology and Microbiology"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>9}, "Psychology"=>{"Psychology"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>1}, "Computer Science"=>{"Computer Science"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>2}, "Unspecified"=>{"Unspecified"=>2}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>2}}, "reader_count_by_country"=>{"Italy"=>1, "Mexico"=>1}, "group_count"=>2}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/2152654"], "description"=>"<p>Effects of NAC on reduced glutathione (GSH) levels in H9c2 cells exposed to CYS9 for 2 hours. (mean + SD from 3 independent experiments). *<i>p</i> < 0.01 compared with control group; §<i>p</i> < 0.01 compared with CYS9 group.</p>", "links"=>[], "tags"=>["nac", "rat liver homogenate", "lactate dehydrogenase release", "candidate cardioprotectant agents", "iso", "ros", "hcy", "electrospray tandem mass spectrometry", "CYS 9", "Reactive oxygen species", "acrolein", "bio", "cyp", "S 9 fraction", "S 9 mix", "mtt", "250 μ m", "CEPM"], "article_id"=>1465984, "categories"=>["Uncategorised"], "users"=>["Takuro Nishikawa", "Emiko Miyahara", "Koichiro Kurauchi", "Erika Watanabe", "Kazuro Ikawa", "Kousuke Asaba", "Takayuki Tanabe", "Yasuhiro Okamoto", "Yoshifumi Kawano"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0131394.g009", "stats"=>{"downloads"=>2, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Reduced_glutathione_levels_in_H9c2_cells_after_treatment_with_CYS9_in_presence_or_absence_of_NAC_/1465984", "title"=>"Reduced glutathione levels in H9c2 cells after treatment with CYS9 in presence or absence of NAC.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-06-26 04:07:26"}
  • {"files"=>["https://ndownloader.figshare.com/files/2152652"], "description"=>"<p>(A) H9c2 cells were exposed for 1 and 2 hours to CYS9 with and without NAC. The changes of acrolein in culture media was measured using HPLC. (mean + SD from 3 independent experiments). Effect of NAC on ROS generated by CYS9, as shown by fluorescence intensity of (B) DCFH, (C) APF, and (D) HPF in cells exposed for 1 hour to CYS9 or CYS9 plus NAC. Fluorescence intensity is shown in arbitrary units. (mean + SD from 3 independent experiments). *<i>p</i> < 0.05 compared with CYS9 group.</p>", "links"=>[], "tags"=>["nac", "rat liver homogenate", "lactate dehydrogenase release", "candidate cardioprotectant agents", "iso", "ros", "hcy", "electrospray tandem mass spectrometry", "CYS 9", "Reactive oxygen species", "acrolein", "bio", "cyp", "S 9 fraction", "S 9 mix", "mtt", "250 μ m", "CEPM"], "article_id"=>1465982, "categories"=>["Uncategorised"], "users"=>["Takuro Nishikawa", "Emiko Miyahara", "Koichiro Kurauchi", "Erika Watanabe", "Kazuro Ikawa", "Kousuke Asaba", "Takayuki Tanabe", "Yasuhiro Okamoto", "Yoshifumi Kawano"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0131394.g007", "stats"=>{"downloads"=>6, "page_views"=>64, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_concentration_of_acrolein_in_cell_culture_media_and_ROS_generation_in_H9c2_cells_after_exposure_to_CYS9_with_and_without_NAC_/1465982", "title"=>"The concentration of acrolein in cell culture media and ROS generation in H9c2 cells after exposure to CYS9 with and without NAC.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-06-26 04:07:26"}
  • {"files"=>["https://ndownloader.figshare.com/files/2152642"], "description"=>"<p>We measured CY and CY metabolites in blood plasma samples from patients receiving high-dose CY and here present the concentration–time profiles obtained from all three patients for (A) CY, (B) HCY, and (C) CEPM. Underlying diseases were acute mixed lineage leukemia (male, 8 y.o.; open square) and granulocytic sarcoma (male, 17 y.o.; open circle), and acute myeloid leukemia (male, 1 y.o.; open triangle).</p>", "links"=>[], "tags"=>["nac", "rat liver homogenate", "lactate dehydrogenase release", "candidate cardioprotectant agents", "iso", "ros", "hcy", "electrospray tandem mass spectrometry", "CYS 9", "Reactive oxygen species", "acrolein", "bio", "cyp", "S 9 fraction", "S 9 mix", "mtt", "250 μ m", "CEPM"], "article_id"=>1465973, "categories"=>["Uncategorised"], "users"=>["Takuro Nishikawa", "Emiko Miyahara", "Koichiro Kurauchi", "Erika Watanabe", "Kazuro Ikawa", "Kousuke Asaba", "Takayuki Tanabe", "Yasuhiro Okamoto", "Yoshifumi Kawano"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0131394.g003", "stats"=>{"downloads"=>2, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Pharmacokinetics_of_high_dose_cyclophosphamide_in_patients_/1465973", "title"=>"Pharmacokinetics of high-dose cyclophosphamide in patients.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-06-26 04:07:26"}
  • {"files"=>["https://ndownloader.figshare.com/files/2152641"], "description"=>"<p>H9c2 cell viability after 24-hour exposure to N-acetylcysteine (NAC) was assessed by MTT assay (mean + standard deviation (SD) from 4 independent experiments). *<i>p</i> < 0.05 compared with control.</p>", "links"=>[], "tags"=>["nac", "rat liver homogenate", "lactate dehydrogenase release", "candidate cardioprotectant agents", "iso", "ros", "hcy", "electrospray tandem mass spectrometry", "CYS 9", "Reactive oxygen species", "acrolein", "bio", "cyp", "S 9 fraction", "S 9 mix", "mtt", "250 μ m", "CEPM"], "article_id"=>1465972, "categories"=>["Uncategorised"], "users"=>["Takuro Nishikawa", "Emiko Miyahara", "Koichiro Kurauchi", "Erika Watanabe", "Kazuro Ikawa", "Kousuke Asaba", "Takayuki Tanabe", "Yasuhiro Okamoto", "Yoshifumi Kawano"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0131394.g002", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Cytotoxicity_of_N_acetylcysteine_on_H9c2_cells_/1465972", "title"=>"Cytotoxicity of N-acetylcysteine on H9c2 cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-06-26 04:07:26"}
  • {"files"=>["https://ndownloader.figshare.com/files/2152656"], "description"=>"<p>H9c2 cell viability after (A) 24-hour and (B) 48-hour exposure to acrolein (Acr) with or without NAC was assessed by MTT assay (mean + SD from 2 independent experiments conducted in duplicate). *<i>p</i> < 0.05 compared with control group. †<i>p</i> < 0.05 compared with acrolein 100 μM group.</p>", "links"=>[], "tags"=>["nac", "rat liver homogenate", "lactate dehydrogenase release", "candidate cardioprotectant agents", "iso", "ros", "hcy", "electrospray tandem mass spectrometry", "CYS 9", "Reactive oxygen species", "acrolein", "bio", "cyp", "S 9 fraction", "S 9 mix", "mtt", "250 μ m", "CEPM"], "article_id"=>1465986, "categories"=>["Uncategorised"], "users"=>["Takuro Nishikawa", "Emiko Miyahara", "Koichiro Kurauchi", "Erika Watanabe", "Kazuro Ikawa", "Kousuke Asaba", "Takayuki Tanabe", "Yasuhiro Okamoto", "Yoshifumi Kawano"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0131394.g010", "stats"=>{"downloads"=>2, "page_views"=>17, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Myocardial_cytotoxicity_induced_by_acrolein_/1465986", "title"=>"Myocardial cytotoxicity induced by acrolein.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-06-26 04:07:26"}
  • {"files"=>["https://ndownloader.figshare.com/files/2152649"], "description"=>"<p>H9c2 cells were exposed to CYS9 for 1 or 2 hours with and without candidate cardioprotectant agents (NAC, ISO, and BIO). Changes in concentration in H9c2 cell culture media of (A) CY and its metabolites (B) HCY and (C) CEPM was evaluated using LC/MS/MS. (mean + SD from 3 independent experiments). *<i>p</i> < 0.05 compared with CYS9 group.</p>", "links"=>[], "tags"=>["nac", "rat liver homogenate", "lactate dehydrogenase release", "candidate cardioprotectant agents", "iso", "ros", "hcy", "electrospray tandem mass spectrometry", "CYS 9", "Reactive oxygen species", "acrolein", "bio", "cyp", "S 9 fraction", "S 9 mix", "mtt", "250 μ m", "CEPM"], "article_id"=>1465980, "categories"=>["Uncategorised"], "users"=>["Takuro Nishikawa", "Emiko Miyahara", "Koichiro Kurauchi", "Erika Watanabe", "Kazuro Ikawa", "Kousuke Asaba", "Takayuki Tanabe", "Yasuhiro Okamoto", "Yoshifumi Kawano"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0131394.g006", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_CY_and_CY_metabolites_results_from_LC_MS_MS_assays_of_culture_supernatants_of_CYS9_with_and_without_candidate_cardioprotectant_agents_/1465980", "title"=>"CY and CY metabolites results from LC/MS/MS assays of culture supernatants of CYS9 with and without candidate cardioprotectant agents.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-06-26 04:07:26"}
  • {"files"=>["https://ndownloader.figshare.com/files/2152646"], "description"=>"<p>(A) The effect of candidate cardioprotectant agents (NAC, isorhamnetin (ISO), and β-ionone (BIO)) on cytotoxicity of CYS9 in H9c2 cells after 24-hour exposure. (mean + SD from 2 independent experiments conducted in duplicate). *<i>p</i> < 0.05 compared with CYS9 group. (B) The effects of candidate cardioprotectant agents against LDH release from H9c2 cells exposed to CYS9 for 2 hours. (mean + SD from 3 independent experiments). *<i>p</i> < 0.05 compared with CYS9 group.</p>", "links"=>[], "tags"=>["nac", "rat liver homogenate", "lactate dehydrogenase release", "candidate cardioprotectant agents", "iso", "ros", "hcy", "electrospray tandem mass spectrometry", "CYS 9", "Reactive oxygen species", "acrolein", "bio", "cyp", "S 9 fraction", "S 9 mix", "mtt", "250 μ m", "CEPM"], "article_id"=>1465977, "categories"=>["Uncategorised"], "users"=>["Takuro Nishikawa", "Emiko Miyahara", "Koichiro Kurauchi", "Erika Watanabe", "Kazuro Ikawa", "Kousuke Asaba", "Takayuki Tanabe", "Yasuhiro Okamoto", "Yoshifumi Kawano"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0131394.g005", "stats"=>{"downloads"=>2, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Inhibition_of_CYS9_induced_cell_cytotoxicity_by_candidate_cardioprotectant_agents_/1465977", "title"=>"Inhibition of CYS9-induced cell cytotoxicity by candidate cardioprotectant agents.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-06-26 04:07:26"}
  • {"files"=>["https://ndownloader.figshare.com/files/2152644"], "description"=>"<p>H9c2 cell viability after (A) 24-hour and (B) 48-hour exposure to CY alone and CY metabolized by S9 fraction of rat liver homogenate mixed with co-factors (CYS9) was assessed by MTT assay (mean + SD from 3 independent experiments). (C) The changes of CY and its metabolites HCY and CEPM concentration in H9c2 cell culture media exposed to CYS9 (mean + SD from 3 independent experiments). (D) Fluorescence intensities, corresponding to levels of H<sub>2</sub>O<sub>2</sub>, in control samples or cells exposed to 250 μM CY, S9, CYS9 for 1 hour (mean + SD from 3 independent experiments). Fluorescence intensity is shown in arbitrary units. *<i>p</i> < 0.05 compared with control.</p>", "links"=>[], "tags"=>["nac", "rat liver homogenate", "lactate dehydrogenase release", "candidate cardioprotectant agents", "iso", "ros", "hcy", "electrospray tandem mass spectrometry", "CYS 9", "Reactive oxygen species", "acrolein", "bio", "cyp", "S 9 fraction", "S 9 mix", "mtt", "250 μ m", "CEPM"], "article_id"=>1465975, "categories"=>["Uncategorised"], "users"=>["Takuro Nishikawa", "Emiko Miyahara", "Koichiro Kurauchi", "Erika Watanabe", "Kazuro Ikawa", "Kousuke Asaba", "Takayuki Tanabe", "Yasuhiro Okamoto", "Yoshifumi Kawano"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0131394.g004", "stats"=>{"downloads"=>2, "page_views"=>32, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Myocardial_cytotoxicity_induced_by_CY_metabolized_by_S9_mix_/1465975", "title"=>"Myocardial cytotoxicity induced by CY metabolized by S9 mix.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-06-26 04:07:26"}
  • {"files"=>["https://ndownloader.figshare.com/files/2152658"], "description"=>"<p>Cell viability was assessed by MTT assay after HL-60 cells were exposed to CYS9 with and without NAC or ISO or BIO: results show cell viability (mean + SD from 3 or 4 independent experiments) after exposure to CYS9 for 24 hours (A, C, E) or 48 hours (B, D, F). *<i>p</i> < 0.05 compared with CYS9 group.</p>", "links"=>[], "tags"=>["nac", "rat liver homogenate", "lactate dehydrogenase release", "candidate cardioprotectant agents", "iso", "ros", "hcy", "electrospray tandem mass spectrometry", "CYS 9", "Reactive oxygen species", "acrolein", "bio", "cyp", "S 9 fraction", "S 9 mix", "mtt", "250 μ m", "CEPM"], "article_id"=>1465988, "categories"=>["Uncategorised"], "users"=>["Takuro Nishikawa", "Emiko Miyahara", "Koichiro Kurauchi", "Erika Watanabe", "Kazuro Ikawa", "Kousuke Asaba", "Takayuki Tanabe", "Yasuhiro Okamoto", "Yoshifumi Kawano"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0131394.g011", "stats"=>{"downloads"=>2, "page_views"=>18, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_CYS9_induced_cytotoxicity_in_HL_60_cells_with_candidate_cardioprotectant_agents_/1465988", "title"=>"CYS9-induced cytotoxicity in HL-60 cells with candidate cardioprotectant agents.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-06-26 04:07:26"}
  • {"files"=>["https://ndownloader.figshare.com/files/2152640"], "description"=>"<p>Cyclophosphamide (CY) is metabolized to 4-hydroxy-cyclophosphamide (HCY) in the hepatic cytochrome P-450 enzyme (CYP) system (CYP2B6 and/or CYP2C19). HCY enters cells as tautomer aldocyclophosphamide (AldoCY). Through β-elimination, AldoCY can be converted to phosphoramide mustard (PM) and acrolein. Alternatively, AldoCY can also be oxidized to the inactive metabolite <i>o</i>-carboxyethylphosphoramide mustard (CEPM) by aldehyde dehydrogenase 1 (ALDH1). Other metabolites include chloroacetaldehyde (CAA), deschloroethyl-cyclophosphamide (DCCY), 4-keto-cyclophosphamide (KetoCY), hydroxypropyl-phosphoramide mustard (HPPM), imino-cyclophosphamide (IminoCY), and glutathionyl-cyclophosphamide (GSCY).</p>", "links"=>[], "tags"=>["nac", "rat liver homogenate", "lactate dehydrogenase release", "candidate cardioprotectant agents", "iso", "ros", "hcy", "electrospray tandem mass spectrometry", "CYS 9", "Reactive oxygen species", "acrolein", "bio", "cyp", "S 9 fraction", "S 9 mix", "mtt", "250 μ m", "CEPM"], "article_id"=>1465970, "categories"=>["Uncategorised"], "users"=>["Takuro Nishikawa", "Emiko Miyahara", "Koichiro Kurauchi", "Erika Watanabe", "Kazuro Ikawa", "Kousuke Asaba", "Takayuki Tanabe", "Yasuhiro Okamoto", "Yoshifumi Kawano"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0131394.g001", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Cyclophosphamide_metabolic_pathways_/1465970", "title"=>"Cyclophosphamide metabolic pathways.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-06-26 04:07:26"}
  • {"files"=>["https://ndownloader.figshare.com/files/2152653"], "description"=>"<p>A, B, C, D: Optical images at 24-hour exposure. (A) Control (unexposed H9c2 cells), (B) H9c2 cells exposed to 250 μM CY, (C) H9c2 cells exposed to CYS9, and (D) H9c2 cells exposed to CYS9 presence of 1 mM NAC. Magnification, 100×. Bar = 200 μm. E, F, G, H: Induction of apoptosis in H9c2 cells by CYS9 with or without NAC. Living cell nucleii stained by Hoechst 33342 are blue. Apoptotic cells stained by FITC-conjugated probes are green. (E) Control (unexposed H9c2 cells), (F) H9c2 cells exposed for 2 hours to 250 μM CY, (G) H9c2 cells exposed to CYS9—green dots indicate apoptotic cells. (H) H9c2 cells exposed to CYS9 with 1 mM of NAC. Magnification, 100×. Bar = 200 μm.</p>", "links"=>[], "tags"=>["nac", "rat liver homogenate", "lactate dehydrogenase release", "candidate cardioprotectant agents", "iso", "ros", "hcy", "electrospray tandem mass spectrometry", "CYS 9", "Reactive oxygen species", "acrolein", "bio", "cyp", "S 9 fraction", "S 9 mix", "mtt", "250 μ m", "CEPM"], "article_id"=>1465983, "categories"=>["Uncategorised"], "users"=>["Takuro Nishikawa", "Emiko Miyahara", "Koichiro Kurauchi", "Erika Watanabe", "Kazuro Ikawa", "Kousuke Asaba", "Takayuki Tanabe", "Yasuhiro Okamoto", "Yoshifumi Kawano"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0131394.g008", "stats"=>{"downloads"=>2, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Optical_and_fluorescence_images_of_H9c2_cells_exposed_to_CY_CYS9_and_CYS9_plus_NAC_/1465983", "title"=>"Optical and fluorescence images of H9c2 cells exposed to CY, CYS9, and CYS9 plus NAC.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-06-26 04:07:26"}

PMC Usage Stats | Further Information

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  • {"unique-ip"=>"10", "full-text"=>"12", "pdf"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"3", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"5"}
  • {"unique-ip"=>"11", "full-text"=>"10", "pdf"=>"4", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"3", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"8"}
  • {"unique-ip"=>"10", "full-text"=>"9", "pdf"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"9"}
  • {"unique-ip"=>"10", "full-text"=>"10", "pdf"=>"5", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"10"}
  • {"unique-ip"=>"13", "full-text"=>"9", "pdf"=>"10", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"12"}

Relative Metric

{"start_date"=>"2015-01-01T00:00:00Z", "end_date"=>"2015-12-31T00:00:00Z", "subject_areas"=>[]}
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