Ultra-Rapid Warming Yields High Survival of Mouse Oocytes Cooled to −196°C in Dilutions of a Standard Vitrification Solution
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{"title"=>"Ultra-rapid warming yields high survival of mouse oocytes cooled to -196°c in dilutions of a standard vitrification solution", "type"=>"journal", "authors"=>[{"first_name"=>"Shinsuke", "last_name"=>"Seki", "scopus_author_id"=>"8833396500"}, {"first_name"=>"Peter", "last_name"=>"Mazur", "scopus_author_id"=>"7103102657"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"sgr"=>"84860449067", "doi"=>"10.1371/journal.pone.0036058", "pui"=>"364720857", "pmid"=>"22558325", "scopus"=>"2-s2.0-84860449067", "issn"=>"19326203", "isbn"=>"1932-6203"}, "id"=>"d4af8277-157c-3942-b130-5afd77d4ea97", "abstract"=>"Intracellular ice is generally lethal. One way to avoid it is to vitrify cells; that is, to convert cell water to a glass rather than to ice. The belief has been that this requires both the cooling rate and the concentration of glass-inducing solutes be very high. But high solute concentrations can themselves be damaging. However, the findings we report here on the vitrification of mouse oocytes are not in accord with the first belief that cooling needs to be extremely rapid. The important requirement is that the warming rate be extremely high. We subjected mouse oocytes in the vitrification solution EAFS 10/10 to vitrification procedures using a broad range of cooling and warming rates. Morphological survivals exceeded 80% when they were warmed at the highest rate (117,000°C/min) even when the prior cooling rate was as low as 880°C/min. Functional survival was >81% and 54% with the highest warming rate after cooling at 69,000 and 880°C/min, respectively. Our findings are also contrary to the second belief. We show that a high percentage of mouse oocytes survive vitrification in media that contain only half the usual concentration of solutes, provided they are warmed extremely rapidly; that is, >100,000°C/min. Again, the cooling rate is of less consequence.", "link"=>"http://www.mendeley.com/research/ultrarapid-warming-yields-high-survival-mouse-oocytes-cooled-196c-dilutions-standard-vitrification-s", "reader_count"=>32, "reader_count_by_academic_status"=>{"Unspecified"=>2, "Professor > Associate Professor"=>3, "Researcher"=>8, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>6, "Student > Postgraduate"=>1, "Student > Master"=>7, "Other"=>1, "Professor"=>3}, "reader_count_by_user_role"=>{"Unspecified"=>2, "Professor > Associate Professor"=>3, "Researcher"=>8, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>6, "Student > Postgraduate"=>1, "Student > Master"=>7, "Other"=>1, "Professor"=>3}, "reader_count_by_subject_area"=>{"Unspecified"=>4, "Engineering"=>3, "Biochemistry, Genetics and Molecular Biology"=>2, "Materials Science"=>1, "Agricultural and Biological Sciences"=>16, "Medicine and Dentistry"=>2, "Veterinary Science and Veterinary Medicine"=>1, "Physics and Astronomy"=>3}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>3}, "Materials Science"=>{"Materials Science"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>2}, "Physics and Astronomy"=>{"Physics and Astronomy"=>3}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>16}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>2}, "Unspecified"=>{"Unspecified"=>4}, "Veterinary Science and Veterinary Medicine"=>{"Veterinary Science and Veterinary Medicine"=>1}}, "reader_count_by_country"=>{"Argentina"=>1, "United States"=>2, "Brazil"=>1}, "group_count"=>1}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/645634"], "description"=>"<p><sup>*</sup>“Morphological survival\" means an oocyte exhibiting normal osmotic responsiveness during the removal of EAFS and exhibiting normal morphology after 2 hr incubation in M16. That is, in the column labeled “+ 2 hr in M16\", after the vitrified oocytes were warmed and the EAFS was removed, the eggs were incubated in M16 medium for 2 hr before being assessed for morphological normalcy. These are the survivals shown graphically in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036058#pone-0036058-g001\" target=\"_blank\">Figures 1</a> and <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036058#pone-0036058-g002\" target=\"_blank\">2</a>. In the column labeled “0 hr in M16\", as soon as the EAFS was removed, the eggs were scored for morphological survival. This was immediately followed by partial dissection of the zonae, and the mixing of eggs and sperm for IVF. In nearly all cases, each treatment was repeated 6 to 9 times with about 6 oocytes per repeat. In column of % of oocytes develop to 2-cell, values with different superscripts were significantly different (P<0.05) by one-way ANOVA. Letters show the differences of survivals with the same warming rate, same diluted vitrification solution and various cooling rates, and there is no significant differences with same warming rate and same cooling rate and various diluted vitrification solutions.</p>†<p>The number of oocytes developing to 2-cell embryos after IVF/the number recovered after vitrification.</p>‡<p>Column 5/Column 4.</p>", "links"=>[], "tags"=>["oocytes", "suspended", "dilution", "cooled", "indicated", "rates", "warmed"], "article_id"=>316128, "categories"=>["Physiology", "Biotechnology"], "users"=>["Shinsuke Seki", "Peter Mazur"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036058.t006", "stats"=>{"downloads"=>1, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Morphological_and_functional_survival_of_mouse_oocytes_suspended_in_various_dilution_of_EAFS10_10_cooled_at_indicated_rates_on_Cryotops_and_warmed_at_117_500_176_C_min_/316128", "title"=>"Morphological and functional survival of mouse oocytes suspended in various dilution of EAFS10/10, cooled at indicated rates on Cryotops, and warmed at 117,500°C/min.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-20 05:48:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/645800"], "description"=>"<p>Columns D–G. The molalities are calculated as Columns N-Q of <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036058#pone-0036058-t002\" target=\"_blank\">Table 2</a>/MW of the respective solute.</p><p>Columns H–K: The molarities are calculated as 100× Column B/Wt% in Column G-K of <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036058#pone-0036058-t002\" target=\"_blank\">Table 2</a>/MW of the solute.</p><p>Column L: The nonpermeating solutes are sucrose and PBS (we ignore the very small contribution of the Ficoll and BSA.) Their combined osmolality is Column F+0.276. We assume the osmolality of PBS to be equal to that of the same molality of NaCl. The osmolality of NaCl = 2φm where 2 is the number of species into which the molecule dissociates and φ is the osmotic coefficient.</p><p>Column M: The volume of water in the oocytes after equilibration with the external medium relative to the volume of water in an isotonic cell. It is 0.276/Column L.</p>", "links"=>[], "tags"=>["solute", "concentrations", "dilutions", "vitrification"], "article_id"=>316293, "categories"=>["Physiology", "Biotechnology"], "users"=>["Shinsuke Seki", "Peter Mazur"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036058.t001", "stats"=>{"downloads"=>1, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Derivative_solute_concentrations_in_various_dilutions_of_the_EAFS10_10_vitrification_solution_/316293", "title"=>"Derivative solute concentrations in various dilutions of the EAFS10/10 vitrification solution.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-20 05:49:37"}
  • {"files"=>["https://ndownloader.figshare.com/files/645569"], "description"=>"<p>The relative concentrations ranged from 0.33× to 1×. The absolute concentrations of the individual solutes are given in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036058#pone-0036058-t001\" target=\"_blank\">Table 1</a> and <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036058#pone-0036058-t002\" target=\"_blank\">2</a>. The suspensions were cooled to −196°C at rates ranging from 522 to 69,250°C/min and subsequently warmed at five different rates. The oocytes in the curves delineated by the open circles and triangles were warmed on Cryotops at the highest attainable rate of 117,500°C/min. The cells in the curves delineated by the closed symbols were warmed at lower rates between 610 and 2,950°C/min. CR and WR stand for cooling rate and warming rate. Symbol CR (°C/min) WR (°C/min) Δ 69,250 117,500 ○ 880 117,500 ▴ 69,250 610 ▪ 522 2,950 ♦ 522 2,170 Survivals are based on morphological appearance and osmotic behavior after 1–2 hr incubation in M16 medium.</p>", "links"=>[], "tags"=>["oocytes", "eafs", "vitrification"], "article_id"=>316071, "categories"=>["Physiology", "Biotechnology"], "users"=>["Shinsuke Seki", "Peter Mazur"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036058.g002", "stats"=>{"downloads"=>2, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Survival_of_mouse_oocytes_vs_the_relative_concentration_of_the_EAFS_vitrification_solution_in_which_they_were_suspended_/316071", "title"=>"Survival of mouse oocytes vs. the relative concentration of the EAFS vitrification solution in which they were suspended.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 05:48:33"}
  • {"files"=>["https://ndownloader.figshare.com/files/645736"], "description"=>"*<p>From <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036058#pone-0036058-t001\" target=\"_blank\">Table 1</a> of <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036058#pone.0036058-Seki2\" target=\"_blank\">[16]</a>.</p><p>Column C: Col. B X density EAFS from <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036058#pone.0036058-Seki2\" target=\"_blank\">[16]</a>.</p><p>Column D: Density of PB1 = 1.003; therefore volume (ml) ≒ mass (g).</p><p>Column F = Column C/Column E.</p><p>Column G = 9.62× Column F, where 9.62 is from <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036058#pone-0036058-t001\" target=\"_blank\">Table 1</a> in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036058#pone.0036058-Seki2\" target=\"_blank\">[16]</a>.</p><p>Columns H-L: Wt% of solutes in 1× EAFS from <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036058#pone-0036058-t001\" target=\"_blank\">Table 1</a> of <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036058#pone.0036058-Seki2\" target=\"_blank\">[16]</a> X Column F.</p><p>Column N = (G/(M/100)*10); Column O = (H/(M/100)*10); etc.</p>", "links"=>[], "tags"=>["compositions", "dilutions", "vitrification"], "article_id"=>316232, "categories"=>["Physiology", "Biotechnology"], "users"=>["Shinsuke Seki", "Peter Mazur"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036058.t002", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Mass_compositions_of_various_dilutions_of_EAFS10_10_vitrification_solution_/316232", "title"=>"Mass compositions of various dilutions of EAFS10/10 vitrification solution.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-20 05:49:21"}
  • {"files"=>["https://ndownloader.figshare.com/files/645767"], "description"=>"*<p>1/4 ml sample straw was covered with a 1/2 ml straw, a 7 mm OD ×90 mm glass tube, and a 10 mm OD ×90 mm glass tube.</p>†<p>The Cryotop was covered with the Cryotop-cap-straw and a 7 mm OD ×90 mm glass tube.</p>‡<p>The straw or Cryotop were placed horizontally on a Styrofoam disk (14 cm diameter, 1.5 cm thick) floated on the surface of LN<sub>2</sub> in a 41 Dewar flask for >5 min before being immersed in LN<sub>2</sub>.</p>§<p>The cooling rates were determined from 20 to −120°C. N = 5.</p>¶<p>The cooling rates were determined in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036058#pone.0036058-Seki1\" target=\"_blank\">[14]</a>.</p>", "links"=>[], "tags"=>["cooling"], "article_id"=>316271, "categories"=>["Physiology", "Biotechnology"], "users"=>["Shinsuke Seki", "Peter Mazur"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036058.t003", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Protocol_of_each_cooling_procedure_/316271", "title"=>"Protocol of each cooling procedure.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-20 05:49:29"}
  • {"files"=>["https://ndownloader.figshare.com/files/645507"], "description"=>"<p>The dilutions were 1.0×, 0.875×, 0.75×, or 0.5×. The cooling rates ranged from 37°C/min to 69,250°C/min; the warming rates (°C/min) were 117,500 (Δ). 2,950 (•). and 2,170 (▪). The protocols are given in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036058#pone-0036058-t003\" target=\"_blank\">Tables 3</a> and <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036058#pone-0036058-t004\" target=\"_blank\">4</a>. Survivals are based on morphological appearance and osmotic behavior after warming and after 1–2 hr incubation in M16 medium.</p>", "links"=>[], "tags"=>["oocytes", "dilution", "eafs", "cooling", "warming"], "article_id"=>315998, "categories"=>["Physiology", "Biotechnology"], "users"=>["Shinsuke Seki", "Peter Mazur"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036058.g001", "stats"=>{"downloads"=>1, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Survival_of_mouse_oocytes_as_a_function_of_the_dilution_of_EAFS_and_the_cooling_and_warming_rate_to_and_from_8722_196_176_C_/315998", "title"=>"Survival of mouse oocytes as a function of the dilution of EAFS and the cooling and warming rate to and from −196°C.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 05:48:13"}
  • {"files"=>["https://ndownloader.figshare.com/files/645706"], "description"=>"<p>The data are % survival ± SEM. The first sets of parentheses are the ratios of the number of surviving oocytes to the number frozen or vitrified. N is the number of replicate. Values with different superscripts were significantly different (P<0.05) by one-way ANOVA. Capital letters shows the differences of survivals with the same warming rate, same diluted vitrification solution and various cooling rate, and small letters shows those with same warming rate and same cooling rate and various diluted vitrification solutions.</p>*<p>shows those with same cooling rate, same dilution of vitrification solution, and different warming rates of 2,170 or 2,950°C/min and <sup>†</sup>shows that it was not possible to do stastical analysis because the other survival was 0%.</p>", "links"=>[], "tags"=>["morphological", "normality", "osmotic", "responsiveness", "oocytes", "suspended", "dilution", "cooled", "warmed", "indicated", "rates", "straws"], "article_id"=>316188, "categories"=>["Physiology", "Biotechnology"], "users"=>["Shinsuke Seki", "Peter Mazur"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036058.t005", "stats"=>{"downloads"=>2, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Survival_based_on_morphological_normality_and_osmotic_responsiveness_of_mouse_oocytes_suspended_in_various_dilution_of_EAFS10_10_and_cooled_and_warmed_at_indicated_rates_in_straws_or_on_Cryotops_/316188", "title"=>"Survival based on morphological normality and osmotic responsiveness of mouse oocytes suspended in various dilution of EAFS10/10, and cooled and warmed at indicated rates in straws or on Cryotops.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-20 05:49:08"}
  • {"files"=>["https://ndownloader.figshare.com/files/645659"], "description"=>"*<p>+, yes; −, no.</p>†<p>The warming rates were calculated between −70 and −35°C in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036058#pone.0036058-Seki2\" target=\"_blank\">[16]</a>.</p>‡<p>The warming rate was determined in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036058#pone.0036058-Kleinhans1\" target=\"_blank\">[18]</a>.</p>", "links"=>[], "tags"=>["warming"], "article_id"=>316158, "categories"=>["Physiology", "Biotechnology"], "users"=>["Shinsuke Seki", "Peter Mazur"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036058.t004", "stats"=>{"downloads"=>1, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Protocol_of_each_warming_procedure_/316158", "title"=>"Protocol of each warming procedure.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-20 05:49:00"}

PMC Usage Stats | Further Information

  • {"scanned-page-browse"=>"0", "month"=>"5", "cited-by"=>"0", "abstract"=>"0", "full-text"=>"25", "unique-ip"=>"27", "pdf"=>"18", "year"=>"2012", "figure"=>"1", "scanned-summary"=>"0", "supp-data"=>"0"}
  • {"month"=>"6", "scanned-page-browse"=>"0", "cited-by"=>"0", "abstract"=>"0", "full-text"=>"26", "year"=>"2012", "pdf"=>"9", "unique-ip"=>"21", "figure"=>"4", "scanned-summary"=>"0", "supp-data"=>"0"}
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  • {"unique-ip"=>"10", "full-text"=>"10", "pdf"=>"6", "abstract"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2012", "month"=>"8"}
  • {"unique-ip"=>"21", "full-text"=>"26", "pdf"=>"8", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"4", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2012", "month"=>"9"}
  • {"unique-ip"=>"27", "full-text"=>"27", "pdf"=>"13", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2012", "month"=>"10"}
  • {"unique-ip"=>"10", "full-text"=>"9", "pdf"=>"7", "abstract"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2012", "month"=>"12"}
  • {"unique-ip"=>"16", "full-text"=>"21", "pdf"=>"6", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2013", "month"=>"1"}
  • {"unique-ip"=>"11", "full-text"=>"9", "pdf"=>"7", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2013", "month"=>"3"}
  • {"unique-ip"=>"10", "full-text"=>"9", "pdf"=>"7", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2013", "month"=>"2"}
  • {"unique-ip"=>"6", "full-text"=>"6", "pdf"=>"5", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2013", "month"=>"4"}
  • {"unique-ip"=>"27", "full-text"=>"30", "pdf"=>"17", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2012", "month"=>"11"}
  • {"unique-ip"=>"5", "full-text"=>"4", "pdf"=>"3", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2013", "month"=>"5"}
  • {"unique-ip"=>"4", "full-text"=>"1", "pdf"=>"3", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2013", "month"=>"6"}
  • {"unique-ip"=>"19", "full-text"=>"17", "pdf"=>"12", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"7", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2013", "month"=>"7"}
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  • {"unique-ip"=>"7", "full-text"=>"5", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2013", "month"=>"10"}
  • {"unique-ip"=>"4", "full-text"=>"5", "pdf"=>"2", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2013", "month"=>"11"}
  • {"unique-ip"=>"6", "full-text"=>"6", "pdf"=>"6", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2013", "month"=>"12"}
  • {"unique-ip"=>"16", "full-text"=>"19", "pdf"=>"6", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2014", "month"=>"1"}
  • {"unique-ip"=>"3", "full-text"=>"4", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2014", "month"=>"2"}
  • {"unique-ip"=>"5", "full-text"=>"5", "pdf"=>"4", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2014", "month"=>"3"}
  • {"unique-ip"=>"11", "full-text"=>"9", "pdf"=>"2", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"4", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2014", "month"=>"5"}
  • {"unique-ip"=>"1", "full-text"=>"1", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2014", "month"=>"6"}
  • {"unique-ip"=>"8", "full-text"=>"7", "pdf"=>"5", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2014", "month"=>"4"}
  • {"unique-ip"=>"6", "full-text"=>"6", "pdf"=>"2", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"4"}
  • {"unique-ip"=>"5", "full-text"=>"7", "pdf"=>"3", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"5"}
  • {"unique-ip"=>"3", "full-text"=>"19", "pdf"=>"0", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"6"}
  • {"unique-ip"=>"8", "full-text"=>"8", "pdf"=>"4", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"7"}
  • {"unique-ip"=>"5", "full-text"=>"5", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"3"}
  • {"unique-ip"=>"11", "full-text"=>"10", "pdf"=>"5", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"2"}
  • {"unique-ip"=>"3", "full-text"=>"2", "pdf"=>"2", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"8"}
  • {"unique-ip"=>"15", "full-text"=>"16", "pdf"=>"7", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"9"}
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Relative Metric

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