137 lines
19 KiB
XML
137 lines
19 KiB
XML
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<mods:title id="FF760C2D7673F25CE6DB6C9F32C805BE">Integrative microscopy to explore physical and nanomechanical properties of eggshells of diapausing embryos in Rotifera: a proof-of-concept study</mods:title>
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<mods:namePart id="D9C1FCE2F1C94A57D299D5F21B668690">Meyer, Stephanie</mods:namePart>
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<mods:namePart id="8954B427627D232311D0FEE0FE12E318">Araujo, Thiago Q.</mods:namePart>
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<mods:namePart id="5F114C0A671FDEB73C1E03105CF512BD">Walsh, Elizabeth J.</mods:namePart>
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<mods:namePart id="1629A957AA5DFF7EAE1C592804C6CFC9">Wallace, Robert L.</mods:namePart>
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<mods:namePart id="1427EF43DE6A3D13789997131C4FFDC8">Hochberg, Rick</mods:namePart>
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<mods:title id="1A0A1018115D7DF6D0C623541CED44DF">Journal of Natural History</mods:title>
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<mods:date id="9461FB64EE5D002EF7D71B4CE56C4D61">2023</mods:date>
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<mods:number id="E64DFADBDA95364C816A534534116226">2023-12-05</mods:number>
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<mods:start id="4804E5FAE11AAB96DD14766A8DC6AD04">1984</mods:start>
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<treatment id="03B687E6220BFFE7FF42FF6FFEF075DE" LSID="urn:lsid:plazi:treatment:03B687E6220BFFE7FF42FF6FFEF075DE" httpUri="http://treatment.plazi.org/id/03B687E6220BFFE7FF42FF6FFEF075DE" lastPageNumber="1991" pageId="7" pageNumber="1991">
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<heading id="D0E8819C220BFFE7FF42FF6FFD6D71D8" bold="true" box="[160,663,142,168]" centered="true" fontSize="11" level="2" pageId="7" pageNumber="1991" reason="3">
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<emphasis id="B96BEAE2220BFFE7FF42FF6FFD6D71D8" bold="true" box="[160,663,142,168]" pageId="7" pageNumber="1991">
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<subSubSection id="C305657B220BFFE7FF42FF6FFE9771D8" box="[160,365,142,168]" pageId="7" pageNumber="1991" type="discussion">
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<paragraph id="8BA036F0220BFFE7FF42FF6FFE9771D8" blockId="7.[160,663,142,168]" box="[160,365,142,168]" pageId="7" pageNumber="1991">
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<emphasis id="B96BEAE2220BFFE7FF42FF6FFE9C71D8" bold="true" box="[160,358,142,168]" italics="true" pageId="7" pageNumber="1991">
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Phylum
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<taxonomicName id="4C1F4D73220BFFE7FEE6FF6FFE9C71D8" ID-CoL="5Y" ID-ENA="10190" box="[260,358,142,168]" kingdom="Animalia" pageId="7" pageNumber="1991" phylum="Rotifera" rank="phylum">Rotifera</taxonomicName>
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</emphasis>
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:
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</paragraph>
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</subSubSection>
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<subSubSection id="C305657B220BFFE7FE97FF6FFD6D71D8" box="[373,663,142,168]" pageId="7" pageNumber="1991" type="nomenclature">
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<paragraph id="8BA036F0220BFFE7FE97FF6FFD6D71D8" blockId="7.[160,663,142,168]" box="[373,663,142,168]" pageId="7" pageNumber="1991">
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<taxonomicName id="4C1F4D73220BFFE7FE97FF6FFD6D71D8" ID-CoL="5X2DW" authority="Pallas, 1776" authorityName="Pallas" authorityYear="1776" box="[373,663,142,168]" class="Eurotatoria" family="Brachionidae" genus="Brachionus" kingdom="Animalia" order="Ploima" pageId="7" pageNumber="1991" phylum="Rotifera" rank="species" species="calyciflorus">Brachionus calyciflorus</taxonomicName>
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</paragraph>
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</subSubSection>
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</emphasis>
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</heading>
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<subSubSection id="C305657B220BFFE7FF42FF23FEF075DE" pageId="7" pageNumber="1991" type="description">
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<paragraph id="8BA036F0220BFFE7FF42FF23FF2871AC" blockId="7.[160,1156,194,393]" box="[160,210,194,220]" pageId="7" pageNumber="1991">
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<emphasis id="B96BEAE2220BFFE7FF42FF23FF2871AC" bold="true" box="[160,210,194,220]" italics="true" pageId="7" pageNumber="1991">SEM</emphasis>
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</paragraph>
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<paragraph id="8BA036F0220BFFE7FF42FF07FD9270F9" blockId="7.[160,1156,194,393]" pageId="7" pageNumber="1991">
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The ovoid, diapausing embryos measured ca 139 µm in diameter
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<emphasis id="B96BEAE2220BFFE7FC71FF07FC52718C" box="[915,936,227,255]" italics="true" pageId="7" pageNumber="1991">(̄x</emphasis>
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= 138.4 ± 5.8 μm). Shells appeared largely devoid of ornamentation at lower magnifications (<1000×), but showed a more rugose surface at higher magnifications (
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<figureCitation id="13242A75220BFFE7FCBAFECAFC247034" box="[856,990,299,324]" captionStart="Figure 1" captionStartId="5.[160,225,1351,1374]" captionTargetBox="[163,1153,142,1314]" captionTargetId="figure-3@5.[163,1153,142,1319]" captionTargetPageId="5" captionText="Figure 1. Characterisation of diapausing embryos of Brachionus calyciflorus. a. Scanning Electron Microscope image of a single egg. b. Close-up of the surface of the egg. c. Ultrastructure of the shell viewed with Transmission Electron Microscopy. d. Close-up of shell ultrastructure. e. Topographic image of the external surface of an egg generated with Atomic Force Microscopy. f. Force–distance (FD) curve generated from a single location on an egg. Parameters for the position of the location on the eggshell are indicated by the cursor positions (software screenshot). Maximum force (MF, green circle) and 20% maximum force (20% MF, orange circle) are indicated on the FD curve. Abbreviations: al, alveolar layer; ar, amorphous layer around embryo; em, embryo; s1, sublayer. Scale bars:A = 25 µm; B = 5 µm; C = 3.5 µm; D = 1.7 µm." figureDoi="http://doi.org/10.5281/zenodo.10479031" httpUri="https://zenodo.org/record/10479031/files/figure.png" pageId="7" pageNumber="1991">Figure 1a,b</figureCitation>
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). Shells often appeared pitted with small pores and craters (0.07–2.96 µm diameter) and occasionally some surface bumps of various textures.
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</paragraph>
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<paragraph id="8BA036F0220BFFE7FF42FE58FF2870A3" blockId="7.[160,1156,441,848]" box="[160,210,441,467]" pageId="7" pageNumber="1991">
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<emphasis id="B96BEAE2220BFFE7FF42FE58FF2870A3" bold="true" box="[160,210,441,467]" italics="true" pageId="7" pageNumber="1991">TEM</emphasis>
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</paragraph>
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<paragraph id="8BA036F0220BFFE7FF42FE3DFF057220" blockId="7.[160,1156,441,848]" pageId="7" pageNumber="1991">
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The shell consisted of two layers – a thick apical layer and a thin basal layer – that together averaged 7.20 ± 1.45 µm (SD) thick (
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<figureCitation id="13242A75220BFFE7FD58FE1EFCDC7368" box="[698,806,511,536]" captionStart="Figure 1" captionStartId="5.[160,225,1351,1374]" captionTargetBox="[163,1153,142,1314]" captionTargetId="figure-3@5.[163,1153,142,1319]" captionTargetPageId="5" captionText="Figure 1. Characterisation of diapausing embryos of Brachionus calyciflorus. a. Scanning Electron Microscope image of a single egg. b. Close-up of the surface of the egg. c. Ultrastructure of the shell viewed with Transmission Electron Microscopy. d. Close-up of shell ultrastructure. e. Topographic image of the external surface of an egg generated with Atomic Force Microscopy. f. Force–distance (FD) curve generated from a single location on an egg. Parameters for the position of the location on the eggshell are indicated by the cursor positions (software screenshot). Maximum force (MF, green circle) and 20% maximum force (20% MF, orange circle) are indicated on the FD curve. Abbreviations: al, alveolar layer; ar, amorphous layer around embryo; em, embryo; s1, sublayer. Scale bars:A = 25 µm; B = 5 µm; C = 3.5 µm; D = 1.7 µm." figureDoi="http://doi.org/10.5281/zenodo.10479031" httpUri="https://zenodo.org/record/10479031/files/figure.png" pageId="7" pageNumber="1991">Figure 1c</figureCitation>
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). The surface of the shell had a wavy appearance and consisted of high ridges and numerous small bumps and valleys. Immediately beneath this was a thick zone of empty pockets that were interconnected through a framework of electron-dense bridges; this layer is described by
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<bibRefCitation id="EF8E4B01220BFFE7FF24FD6BFE4A73D3" author="Wurdak E & Gilbert JJ & Jagles R" box="[198,432,650,675]" pageId="7" pageNumber="1991" pagination="49 - 72" refId="ref12780" refString="Wurdak E, Gilbert JJ, Jagles R. 1978. Fine structure of the resting eggs of the rotifers Brachionus calyciflorus and Asplanchna sieboIdi. Trans Am Microscop Soc. 97 (1): 49 - 72. doi: 10.2307 / 3225684." type="journal article" year="1978">
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Wurdak
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<emphasis id="B96BEAE2220BFFE7FEC5FD6BFEA073D3" box="[295,346,650,675]" italics="true" pageId="7" pageNumber="1991">et al</emphasis>
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. (1978)
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</bibRefCitation>
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as the alveolar layer (al:
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<figureCitation id="13242A75220BFFE7FD3BFD6BFCBD73D3" box="[729,839,650,675]" captionStart="Figure 1" captionStartId="5.[160,225,1351,1374]" captionTargetBox="[163,1153,142,1314]" captionTargetId="figure-3@5.[163,1153,142,1319]" captionTargetPageId="5" captionText="Figure 1. Characterisation of diapausing embryos of Brachionus calyciflorus. a. Scanning Electron Microscope image of a single egg. b. Close-up of the surface of the egg. c. Ultrastructure of the shell viewed with Transmission Electron Microscopy. d. Close-up of shell ultrastructure. e. Topographic image of the external surface of an egg generated with Atomic Force Microscopy. f. Force–distance (FD) curve generated from a single location on an egg. Parameters for the position of the location on the eggshell are indicated by the cursor positions (software screenshot). Maximum force (MF, green circle) and 20% maximum force (20% MF, orange circle) are indicated on the FD curve. Abbreviations: al, alveolar layer; ar, amorphous layer around embryo; em, embryo; s1, sublayer. Scale bars:A = 25 µm; B = 5 µm; C = 3.5 µm; D = 1.7 µm." figureDoi="http://doi.org/10.5281/zenodo.10479031" httpUri="https://zenodo.org/record/10479031/files/figure.png" pageId="7" pageNumber="1991">Figure 1d</figureCitation>
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). Immediately beneath the alveolar layer was a zone [s1:
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<bibRefCitation id="EF8E4B01220BFFE7FE16FD4DFD2273B5" author="Wurdak E & Gilbert JJ & Jagles R" box="[500,728,684,709]" pageId="7" pageNumber="1991" pagination="49 - 72" refId="ref12780" refString="Wurdak E, Gilbert JJ, Jagles R. 1978. Fine structure of the resting eggs of the rotifers Brachionus calyciflorus and Asplanchna sieboIdi. Trans Am Microscop Soc. 97 (1): 49 - 72. doi: 10.2307 / 3225684." type="journal article" year="1978">
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Wurdak
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<emphasis id="B96BEAE2220BFFE7FDB1FD4DFD7E73B5" box="[595,644,684,709]" italics="true" pageId="7" pageNumber="1991">et al</emphasis>
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. (1978)
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</bibRefCitation>
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] of less opaque electron density that consisted of two subzones, each containing many fine laminae, and separated by an electron-dense line. An extraembryonic space of variable size separated the physical shell from an amorphous region (ar) that was proximal to the plasma membrane of the embryo.
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</paragraph>
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<paragraph id="8BA036F0220BFFE7FF42FC9EFF2F72E9" blockId="7.[160,1156,895,1198]" box="[160,213,895,921]" pageId="7" pageNumber="1991">
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<emphasis id="B96BEAE2220BFFE7FF42FC9EFF2F72E9" bold="true" box="[160,213,895,921]" italics="true" pageId="7" pageNumber="1991">AFM</emphasis>
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</paragraph>
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<paragraph id="8BA036F0220BFFE7FF42FC42FEF075DE" blockId="7.[160,1156,895,1198]" pageId="7" pageNumber="1991">
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Topography of the outer shell layer was similar to that viewed with SEM, but pores were less visible
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<emphasis id="B96BEAE2220BFFE7FE8EFC24FE7B72AC" box="[364,385,963,990]" italics="true" pageId="7" pageNumber="1991">(̄x</emphasis>
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= 0.23 µm diameter) and surface bumps were less recognisable (
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<figureCitation id="13242A75220BFFE7FF4AFC09FEEE7571" box="[168,276,1000,1025]" captionStart="Figure 1" captionStartId="5.[160,225,1351,1374]" captionTargetBox="[163,1153,142,1314]" captionTargetId="figure-3@5.[163,1153,142,1319]" captionTargetPageId="5" captionText="Figure 1. Characterisation of diapausing embryos of Brachionus calyciflorus. a. Scanning Electron Microscope image of a single egg. b. Close-up of the surface of the egg. c. Ultrastructure of the shell viewed with Transmission Electron Microscopy. d. Close-up of shell ultrastructure. e. Topographic image of the external surface of an egg generated with Atomic Force Microscopy. f. Force–distance (FD) curve generated from a single location on an egg. Parameters for the position of the location on the eggshell are indicated by the cursor positions (software screenshot). Maximum force (MF, green circle) and 20% maximum force (20% MF, orange circle) are indicated on the FD curve. Abbreviations: al, alveolar layer; ar, amorphous layer around embryo; em, embryo; s1, sublayer. Scale bars:A = 25 µm; B = 5 µm; C = 3.5 µm; D = 1.7 µm." figureDoi="http://doi.org/10.5281/zenodo.10479031" httpUri="https://zenodo.org/record/10479031/files/figure.png" pageId="7" pageNumber="1991">Figure 1e</figureCitation>
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). An example force–distance curve is provided (
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<figureCitation id="13242A75220BFFE7FCA3FC09FC527571" box="[833,936,1000,1025]" captionStart="Figure 1" captionStartId="5.[160,225,1351,1374]" captionTargetBox="[163,1153,142,1314]" captionTargetId="figure-3@5.[163,1153,142,1319]" captionTargetPageId="5" captionText="Figure 1. Characterisation of diapausing embryos of Brachionus calyciflorus. a. Scanning Electron Microscope image of a single egg. b. Close-up of the surface of the egg. c. Ultrastructure of the shell viewed with Transmission Electron Microscopy. d. Close-up of shell ultrastructure. e. Topographic image of the external surface of an egg generated with Atomic Force Microscopy. f. Force–distance (FD) curve generated from a single location on an egg. Parameters for the position of the location on the eggshell are indicated by the cursor positions (software screenshot). Maximum force (MF, green circle) and 20% maximum force (20% MF, orange circle) are indicated on the FD curve. Abbreviations: al, alveolar layer; ar, amorphous layer around embryo; em, embryo; s1, sublayer. Scale bars:A = 25 µm; B = 5 µm; C = 3.5 µm; D = 1.7 µm." figureDoi="http://doi.org/10.5281/zenodo.10479031" httpUri="https://zenodo.org/record/10479031/files/figure.png" pageId="7" pageNumber="1991">Figure 1f</figureCitation>
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) that displays how Young̾s modulus values were collected for a single spot on
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<specimenCount id="9D19FD79220BFFE7FC9CFBEAFC197554" box="[894,995,1035,1060]" count="1" pageId="7" pageNumber="1991" type="egg">one egg</specimenCount>
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. The Young̾s modulus for two diapausing embryos ranged from 9.73 to 17.21 MPa with an average of 13.62 ± 2.43 MPa (SD) (
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<tableCitation id="C69D034B220BFFE7FE0FFBB1FDBD7519" box="[493,583,1104,1129]" captionStart="Table 1" captionStartId="6.[317,373,142,165]" captionTargetPageId="6" captionText="Table 1. Young̍s modulus (MPa) of elasticity values for diapausing" httpUri="http://table.plazi.org/id/DF606678220AFFE6FEDFFF6FFDD5706A" pageId="7" pageNumber="1991" tableUuid="DF606678220AFFE6FEDFFF6FFDD5706A">Table 1</tableCitation>
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). Hardness values ranged from 1.62 × 10−2 to 2.18 × 10−2 GPa with an average hardness of 1.80 × 10−2 GPa ± 1.80 × 10−3 GPa (
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<tableCitation id="C69D034B220BFFE7FF4AFB74FF0775DE" box="[168,253,1173,1198]" captionStart="Table 2" captionStartId="6.[320,376,802,825]" captionTargetBox="[330,985,955,1362]" captionTargetPageId="6" captionText="Table 2. Hardness values (GPa × 10−2) for diapausing embryos of" httpUri="http://table.plazi.org/id/DF606678220AFFE6FEA2FCC3FE2472DE" pageId="7" pageNumber="1991" tableUuid="DF606678220AFFE6FEA2FCC3FE2472DE">Table 2</tableCitation>
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).
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</paragraph>
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</subSubSection>
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</treatment>
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</document> |