05e20cf6c1eaed21d047ac38748e5c20 antioxidants-12-01945.pdf 745d3d91bf55f3eadd0bb4ac00d1d09a313878ab antioxidants-12-01945.pdf 5e38b19d3069e2f33a4296d60c54bfc9d470c96887064f4c171d2133a08102ea antioxidants-12-01945.pdf Title: Targeting Pro-Oxidant Iron with Exogenously Administered Apotransferrin Provides Benefits Associated with Changes in Crucial Cellular Iron Gate Protein TfR in a Model of Intracerebral Hemorrhagic Stroke in Mice Subject: We have previously demonstrated that the post-stroke administration of iron-free transferrin (apotransferrin, ATf) is beneficial in different models of ischemic stroke (IS) through the inhibition of the neuronal uptake of pro-oxidant iron. In the present study, we asked whether ATf is safe and also beneficial when given after the induction of intracerebral hemorrhage (ICH) in mice, and investigated the underlying mechanisms. We first compared the main iron actors in the brain of IS- or collagenase-induced ICH mice and then obtained insight into these iron-related proteins in ICH 72 h after the administration of ATf. The infarct size of the IS mice was double that of hemorrhage in ICH mice, but both groups showed similar body weight loss, edema, and increased ferritin and transferrin levels in the ipsilateral brain hemisphere. Although the administration of human ATf (hATf) to ICH mice did not alter the hemorrhage volume or levels of the classical ferroptosis GPX4/system xc- pathways, hATf induced better neurobehavioral performance, decreased 4-hydroxynonenal levels and those of the second-generation ferroptosis marker transferrin receptor (TfR), and restored the mRNA levels of the recently recognized cytosolic iron chaperone poly(RC) binding protein 2. In addition, hATf treatment lowered the ICH-induced increase in both endogenous mouse transferrin mRNA levels and the activation of caspase-2. In conclusion, hATf treatment provides neurobehavioral benefits post-ICH associated with the modulation of iron/oxidative players. Keywords: ischemic stroke; intracerebral hemorrhage; iron; oxidation; apotransferrin; mouse Author: Alexia García-Serran, Jesús Ordoño, Núria DeGregorio-Rocasolano, Marc Melià-Sorolla, Karla Odendaal, Octavi Martí-Sistac and Teresa Gasull Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Tue Oct 31 11:08:29 2023 CET ModDate: Tue Oct 31 11:51:20 2023 CET Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 18 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 4155083 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- MWHIBA+VnURWPalladioL Type 1 Custom yes yes no 10 0 CTJUIH+URWPalladioL-Bold Type 1 Custom yes yes no 15 0 RCVJPV+URWPalladioL-Roma Type 1 Custom yes yes no 20 0 WMCUPI+URWPalladioL-Ital Type 1 Custom yes yes no 24 0 EMHZMG+VnURWPalladioL-Bold Type 1 Custom yes yes no 28 0 KUDXZB+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 46 0 XFIZIB+CMSY10 Type 1 Builtin yes yes no 68 0 CAMXTK+EURM10 Type 1 Builtin yes yes no 75 0 MPUDFL+PazoMath Type 1 Builtin yes yes no 82 0 ZPGVCH+CMEX10 Type 1 Builtin yes yes no 89 0 GIGFZE+CMR10 Type 1 Builtin yes yes no 93 0 JGPGBB+PalatinoLinotype,Italic TrueType MacRoman yes yes no 103 0 JGPCMN+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 106 0 JGPCMO+PalatinoLinotype CID TrueType Identity-H yes yes yes 109 0 JGPCMP+PalatinoLinotype TrueType WinAnsi yes yes no 115 0 JGPCNB+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 118 0 JGPEDK+PalatinoLinotype TrueType MacRoman yes yes no 121 0 JGPCMN+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 134 0 JGPCMO+PalatinoLinotype CID TrueType Identity-H yes yes yes 137 0 JGPCMP+PalatinoLinotype TrueType WinAnsi yes yes no 143 0 JGPCNB+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 146 0 JGPEDK+PalatinoLinotype TrueType MacRoman yes yes no 149 0 JGPCMN+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 162 0 JGPCMO+PalatinoLinotype CID TrueType Identity-H yes yes yes 165 0 JGPCMP+PalatinoLinotype TrueType WinAnsi yes yes no 171 0 JGPCNB+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 174 0 JGPEDK+PalatinoLinotype TrueType MacRoman yes yes no 177 0 JGPCMN+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 188 0 JGPCMO+PalatinoLinotype CID TrueType Identity-H yes yes yes 191 0 JGPCMP+PalatinoLinotype TrueType WinAnsi yes yes no 197 0 JGPCNA+PalatinoLinotype,Italic CID TrueType Identity-H yes yes yes 200 0 JGPCNB+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 206 0 JGPEDK+PalatinoLinotype TrueType MacRoman yes yes no 209 0 JGPCLM+PalatinoLinotype,Bold CID TrueType Identity-H yes yes yes 221 0 JGPCMN+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 227 0 JGPCMO+PalatinoLinotype CID TrueType Identity-H yes yes yes 230 0 JGPCMP+PalatinoLinotype TrueType WinAnsi yes yes no 236 0 JGPCNA+PalatinoLinotype,Italic CID TrueType Identity-H yes yes yes 239 0 JGPCNB+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 245 0 JGPEDK+PalatinoLinotype TrueType MacRoman yes yes no 248 0 JGPCLM+PalatinoLinotype,Bold CID TrueType Identity-H yes yes yes 259 0 JGPCMN+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 265 0 JGPCMO+PalatinoLinotype CID TrueType Identity-H yes yes yes 268 0 JGPCMP+PalatinoLinotype TrueType WinAnsi yes yes no 274 0 JGPCNA+PalatinoLinotype,Italic CID TrueType Identity-H yes yes yes 277 0 JGPCNB+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 283 0 JGPEDK+PalatinoLinotype TrueType MacRoman yes yes no 286 0 JGPCMN+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 298 0 JGPCMO+PalatinoLinotype CID TrueType Identity-H yes yes yes 301 0 JGPCMP+PalatinoLinotype TrueType WinAnsi yes yes no 307 0 JGPCNB+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 310 0 JGPCMN+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 325 0 JGPCMO+PalatinoLinotype CID TrueType Identity-H yes yes yes 328 0 JGPCMP+PalatinoLinotype TrueType WinAnsi yes yes no 334 0 JGPCNB+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 337 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2023-11-18 02:43:08 CET RepresentationInformation: antioxidants-12-01945.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2023-11-08 14:44:07 CET Size: 4155083 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 574 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4d3167f4 Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@ed9d034 Children: Item: Title: Animals Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@6121c9d6 Item: Title: Hypoxic/Ischemic Stroke Model (IS) Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@87f383f Item: Title: Intracerebral Hemorrhage Stroke Model (ICH) Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4eb7f003 Item: Title: Tape Removal Test Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@eafc191 Item: Title: Tail Bleeding Test Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@612fc6eb Item: Title: Blood and Brain Sampling and Processing Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1060b431 Item: Title: Assessment of the Ischemic Infarct Volume in the IS Model, Hemorrhage Volume, and Brain Hemoglobin and Heme Concentration in the ICH Model Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@612679d6 Item: Title: RT-qPCR Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@11758f2a Item: Title: Western Blot (WB) Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@e720b71 Item: Title: Determination of 4-Hydroxynonenal (4-HNE) Using Immunohistochemistry (IHC) Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1b26f7b2 Item: Title: TSAT Assessment Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@491cc5c9 Item: Title: Antibodies Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@74ad1f1f Item: Title: Statistics Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@6a1aab78 Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@462d5aee Children: Item: Title: Gross Differences and Commonalities between Ischemic and Hemorrhagic Stroke Models Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@69b0fd6f Item: Title: Ischemic and Hemorrhagic Stroke Share a Common Profile of Iron-Binding, Ferroptosis-Related Proteins in the Ipsilateral Brain Hemisphere 72 h after Stroke Induction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@757942a1 Item: Title: Early Peripheral Administration of hATf into the Bloodstream after ICH Induction Does Not Reduce Parenchymal Hemorrhage Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4a87761d Item: Title: hATf Reduces ICH-Induced Sensorimotor Impairment in the Adhesive Tape Detection and Removal Test Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@66d1af89 Item: Title: Effect of hATf on Iron Storage/Transport-Related mRNA and Protein Levels in ICH Mice Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@8646db9 Item: Title: Effects of hATf on Classical Key Ferroptosis mRNA and Protein Levels in ICH Mice Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@37374a5e Item: Title: Effect of hATf on the mRNA Expression of Newly Reported Key Ferroptosis Players AIFM2/FSP1 and PCBP2 in ICH Mice Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4671e53b Item: Title: hATf Reduces ICH-Induced Caspase 2 Activation Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@2db7a79b Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@6950e31 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@b7dd107 Info: Title: Targeting Pro-Oxidant Iron with Exogenously Administered Apotransferrin Provides Benefits Associated with Changes in Crucial Cellular Iron Gate Protein TfR in a Model of Intracerebral Hemorrhagic Stroke in Mice Author: Alexia García-Serran, Jesús Ordoño, Núria DeGregorio-Rocasolano, Marc Melià-Sorolla, Karla Odendaal, Octavi Martí-Sistac and Teresa Gasull Subject: We have previously demonstrated that the post-stroke administration of iron-free transferrin (apotransferrin, ATf) is beneficial in different models of ischemic stroke (IS) through the inhibition of the neuronal uptake of pro-oxidant iron. In the present study, we asked whether ATf is safe and also beneficial when given after the induction of intracerebral hemorrhage (ICH) in mice, and investigated the underlying mechanisms. We first compared the main iron actors in the brain of IS- or collagenase-induced ICH mice and then obtained insight into these iron-related proteins in ICH 72 h after the administration of ATf. The infarct size of the IS mice was double that of hemorrhage in ICH mice, but both groups showed similar body weight loss, edema, and increased ferritin and transferrin levels in the ipsilateral brain hemisphere. Although the administration of human ATf (hATf) to ICH mice did not alter the hemorrhage volume or levels of the classical ferroptosis GPX4/system xc- pathways, hATf induced better neurobehavioral performance, decreased 4-hydroxynonenal levels and those of the second-generation ferroptosis marker transferrin receptor (TfR), and restored the mRNA levels of the recently recognized cytosolic iron chaperone poly(RC) binding protein 2. In addition, hATf treatment lowered the ICH-induced increase in both endogenous mouse transferrin mRNA levels and the activation of caspase-2. In conclusion, hATf treatment provides neurobehavioral benefits post-ICH associated with the modulation of iron/oxidative players. Keywords: ischemic stroke; intracerebral hemorrhage; iron; oxidation; apotransferrin; mouse Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Tue Oct 31 11:08:29 CET 2023 ModDate: Tue Oct 31 11:51:20 CET 2023 ID: 0x6946edb742a447b438778f6b5647fb27, 0x6946edb742a447b438778f6b5647fb27 Filters: FilterPipeline: FlateDecode Fonts: Type1: Font: BaseFont: MPUDFL+PazoMath FontSubset: true FirstChar: 68 LastChar: 68 FontDescriptor: FontName: MPUDFL+PazoMath Flags: Symbolic FontBBox: -40, -283, 878, 946 FontFile: true Font: BaseFont: RCVJPV+URWPalladioL-Roma FontSubset: true FirstChar: 2 LastChar: 252 FontDescriptor: FontName: RCVJPV+URWPalladioL-Roma Flags: Symbolic FontBBox: -166, -283, 1021, 943 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: XFIZIB+CMSY10 FontSubset: true FirstChar: 0 LastChar: 14 FontDescriptor: FontName: XFIZIB+CMSY10 Flags: Symbolic FontBBox: -29, -960, 1116, 775 FontFile: true Font: BaseFont: WMCUPI+URWPalladioL-Ital FontSubset: true FirstChar: 2 LastChar: 121 FontDescriptor: FontName: WMCUPI+URWPalladioL-Ital Flags: Symbolic FontBBox: -170, -305, 1010, 941 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: ZPGVCH+CMEX10 FontSubset: true FirstChar: 18 LastChar: 19 FontDescriptor: FontName: ZPGVCH+CMEX10 Flags: Symbolic FontBBox: -24, -2960, 1454, 772 FontFile: true Font: BaseFont: MWHIBA+VnURWPalladioL FontSubset: true FirstChar: 161 LastChar: 226 FontDescriptor: FontName: MWHIBA+VnURWPalladioL Flags: Symbolic FontBBox: -166, -283, 1021, 943 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: CAMXTK+EURM10 FontSubset: true FirstChar: 12 LastChar: 22 FontDescriptor: FontName: CAMXTK+EURM10 Flags: Symbolic FontBBox: -32, -243, 1060, 720 FontFile: true Font: BaseFont: EMHZMG+VnURWPalladioL-Bold FontSubset: true FirstChar: 160 LastChar: 243 FontDescriptor: FontName: EMHZMG+VnURWPalladioL-Bold Flags: Symbolic FontBBox: -152, -301, 1000, 935 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: GIGFZE+CMR10 FontSubset: true FirstChar: 40 LastChar: 61 FontDescriptor: FontName: GIGFZE+CMR10 Flags: Symbolic FontBBox: -40, -250, 1009, 750 FontFile: true Font: BaseFont: CTJUIH+URWPalladioL-Bold FontSubset: true FirstChar: 2 LastChar: 241 FontDescriptor: FontName: CTJUIH+URWPalladioL-Bold Flags: Symbolic FontBBox: -152, -301, 1000, 935 FontFile: true EncodingDictionary: Differences: true Pages: Page: Sequence: 1 Annotations: Annotation: Subtype: Link Rect: 34, 291, 152, 300 Annotation: Subtype: Link Rect: 34, 281, 83, 288 Annotation: Subtype: Link Rect: 34, 164, 96, 186 Annotation: Subtype: Link Rect: 126, 90, 154, 99 Annotation: Subtype: Link Rect: 34, 78, 147, 87 Annotation: Subtype: Link Rect: 34, 756, 175, 792 Annotation: Subtype: Link Rect: 515, 756, 560, 792 Annotation: Subtype: Link Rect: 136, 623, 147, 636 Annotation: Subtype: Link Rect: 368, 623, 380, 636 Annotation: Subtype: Link Rect: 478, 623, 489, 636 Annotation: Subtype: Link Rect: 330, 611, 341, 624 Annotation: Subtype: Link Rect: 480, 158, 487, 167 ActionDest: 348 Annotation: Subtype: Link Rect: 490, 158, 497, 167 ActionDest: 351 Annotation: Subtype: Link Rect: 235, 145, 241, 154 ActionDest: 351 Annotation: Subtype: Link Rect: 251, 120, 258, 129 ActionDest: 351 Annotation: Subtype: Link Rect: 234, 108, 241, 117 ActionDest: 351 Annotation: Subtype: Link Rect: 242, 108, 249, 117 ActionDest: 351 Annotation: Subtype: Link Rect: 243, 70, 250, 79 ActionDest: 351 Annotation: Subtype: Link Rect: 131, 17, 277, 28 Annotation: Subtype: Link Rect: 392, 17, 560, 28 Annotation: Subtype: Link Rect: 34, 436, 87, 455 Page: Sequence: 2 Annotations: Annotation: Subtype: Link Rect: 528, 710, 535, 719 ActionDest: 351 Annotation: Subtype: Link Rect: 535, 710, 542, 719 ActionDest: 351 Annotation: Subtype: Link Rect: 529, 685, 541, 694 ActionDest: 351 Annotation: Subtype: Link Rect: 544, 685, 556, 694 ActionDest: 351 Annotation: Subtype: Link Rect: 238, 647, 249, 657 ActionDest: 351 Annotation: Subtype: Link Rect: 250, 648, 262, 657 ActionDest: 351 Annotation: Subtype: Link Rect: 503, 635, 515, 644 ActionDest: 351 Annotation: Subtype: Link Rect: 235, 610, 247, 619 ActionDest: 351 Annotation: Subtype: Link Rect: 529, 597, 541, 606 ActionDest: 351 Annotation: Subtype: Link Rect: 544, 597, 556, 606 ActionDest: 351 Annotation: Subtype: Link Rect: 386, 484, 398, 493 ActionDest: 351 Annotation: Subtype: Link Rect: 388, 182, 400, 191 ActionDest: 351 Page: Sequence: 3 Annotations: Annotation: Subtype: Link Rect: 417, 720, 429, 729 ActionDest: 351 Annotation: Subtype: Link Rect: 449, 79, 461, 88 ActionDest: 351 Page: Sequence: 4 Annotations: Annotation: Subtype: Link Rect: 430, 574, 442, 583 ActionDest: 351 Page: Sequence: 5 Annotations: Annotation: Subtype: Link Rect: 492, 710, 504, 719 ActionDest: 351 Annotation: Subtype: Link Rect: 505, 710, 517, 719 ActionDest: 351 Annotation: Subtype: Link Rect: 417, 170, 424, 179 ActionDest: 351 Annotation: Subtype: Link Rect: 425, 170, 437, 179 ActionDest: 351 Page: Sequence: 6 Annotations: Annotation: Subtype: Link Rect: 357, 556, 364, 568 ActionDest: 97 Annotation: Subtype: Link Rect: 420, 543, 427, 555 ActionDest: 97 Annotation: Subtype: Link Rect: 231, 530, 238, 542 ActionDest: 97 Annotation: Subtype: Link Rect: 257, 518, 264, 530 ActionDest: 97 Page: Sequence: 7 Annotations: Annotation: Subtype: Link Rect: 422, 682, 429, 694 ActionDest: 126 Annotation: Subtype: Link Rect: 526, 657, 533, 669 ActionDest: 126 Annotation: Subtype: Link Rect: 477, 644, 484, 656 ActionDest: 126 Annotation: Subtype: Link Rect: 530, 622, 542, 631 ActionDest: 351 Page: Sequence: 8 Annotations: Annotation: Subtype: Link Rect: 187, 672, 194, 681 ActionDest: 351 Annotation: Subtype: Link Rect: 395, 634, 402, 643 ActionDest: 351 Annotation: Subtype: Link Rect: 218, 619, 225, 631 ActionDest: 156 Annotation: Subtype: Link Rect: 262, 559, 269, 568 ActionDest: 351 Annotation: Subtype: Link Rect: 219, 518, 226, 530 ActionDest: 156 Annotation: Subtype: Link Rect: 296, 506, 303, 518 ActionDest: 156 Annotation: Subtype: Link Rect: 474, 506, 481, 518 ActionDest: 156 Annotation: Subtype: Link Rect: 530, 468, 537, 480 ActionDest: 156 Annotation: Subtype: Link Rect: 433, 456, 440, 468 ActionDest: 156 Annotation: Subtype: Link Rect: 221, 443, 228, 455 ActionDest: 156 Annotation: Subtype: Link Rect: 543, 395, 550, 407 ActionDest: 182 Annotation: Subtype: Link Rect: 268, 382, 275, 394 ActionDest: 182 Annotation: Subtype: Link Rect: 305, 282, 312, 294 ActionDest: 182 Annotation: Subtype: Link Rect: 275, 244, 282, 256 ActionDest: 213 Annotation: Subtype: Link Rect: 412, 118, 418, 130 ActionDest: 253 Annotation: Subtype: Link Rect: 341, 93, 348, 105 ActionDest: 253 Page: Sequence: 9 Page: Sequence: 10 Annotations: Annotation: Subtype: Link Rect: 254, 161, 261, 173 ActionDest: 253 Annotation: Subtype: Link Rect: 418, 136, 425, 148 ActionDest: 253 Page: Sequence: 11 Annotations: Annotation: Subtype: Link Rect: 319, 159, 326, 171 ActionDest: 315 Page: Sequence: 12 Page: Sequence: 13 Annotations: Annotation: Subtype: Link Rect: 549, 333, 556, 345 ActionDest: 182 Annotation: Subtype: Link Rect: 269, 258, 276, 270 ActionDest: 97 Annotation: Subtype: Link Rect: 386, 195, 393, 207 ActionDest: 126 Annotation: Subtype: Link Rect: 330, 160, 337, 169 ActionDest: 351 Annotation: Subtype: Link Rect: 338, 160, 350, 169 ActionDest: 351 Annotation: Subtype: Link Rect: 350, 160, 362, 169 ActionDest: 354 Annotation: Subtype: Link Rect: 365, 160, 377, 169 ActionDest: 354 Annotation: Subtype: Link Rect: 286, 122, 293, 131 ActionDest: 351 Annotation: Subtype: Link Rect: 293, 122, 305, 131 ActionDest: 354 Annotation: Subtype: Link Rect: 324, 110, 331, 119 ActionDest: 351 Annotation: Subtype: Link Rect: 429, 85, 436, 94 ActionDest: 351 Annotation: Subtype: Link Rect: 437, 57, 444, 69 ActionDest: 156 Page: Sequence: 14 Annotations: Annotation: Subtype: Link Rect: 373, 720, 380, 732 ActionDest: 156 Annotation: Subtype: Link Rect: 469, 695, 476, 707 ActionDest: 182 Annotation: Subtype: Link Rect: 330, 660, 337, 669 ActionDest: 351 Annotation: Subtype: Link Rect: 378, 635, 385, 644 ActionDest: 351 Annotation: Subtype: Link Rect: 540, 595, 547, 607 ActionDest: 182 Annotation: Subtype: Link Rect: 424, 419, 431, 431 ActionDest: 156 Annotation: Subtype: Link Rect: 434, 394, 441, 406 ActionDest: 156 Annotation: Subtype: Link Rect: 380, 283, 392, 293 ActionDest: 354 Annotation: Subtype: Link Rect: 392, 283, 404, 293 ActionDest: 354 Annotation: Subtype: Link Rect: 362, 271, 374, 280 ActionDest: 354 Annotation: Subtype: Link Rect: 233, 155, 240, 167 ActionDest: 253 Annotation: Subtype: Link Rect: 321, 143, 329, 155 ActionDest: 253 Annotation: Subtype: Link Rect: 531, 133, 543, 142 ActionDest: 354 Annotation: Subtype: Link Rect: 544, 133, 556, 142 ActionDest: 354 Annotation: Subtype: Link Rect: 388, 95, 400, 104 ActionDest: 354 Page: Sequence: 15 Annotations: Annotation: Subtype: Link Rect: 548, 733, 555, 745 ActionDest: 253 Annotation: Subtype: Link Rect: 412, 698, 424, 707 ActionDest: 351 Annotation: Subtype: Link Rect: 425, 698, 437, 707 ActionDest: 354 Annotation: Subtype: Link Rect: 522, 673, 534, 682 ActionDest: 354 Annotation: Subtype: Link Rect: 338, 635, 350, 644 ActionDest: 354 Annotation: Subtype: Link Rect: 351, 635, 363, 644 ActionDest: 354 Annotation: Subtype: Link Rect: 192, 597, 204, 606 ActionDest: 354 Annotation: Subtype: Link Rect: 489, 572, 501, 581 ActionDest: 351 Annotation: Subtype: Link Rect: 502, 572, 514, 581 ActionDest: 351 Annotation: Subtype: Link Rect: 460, 560, 472, 569 ActionDest: 351 Annotation: Subtype: Link Rect: 473, 560, 485, 569 ActionDest: 354 Annotation: Subtype: Link Rect: 488, 560, 500, 569 ActionDest: 354 Annotation: Subtype: Link Rect: 356, 472, 368, 481 ActionDest: 354 Annotation: Subtype: Link Rect: 479, 459, 491, 468 ActionDest: 354 Annotation: Subtype: Link Rect: 377, 371, 389, 380 ActionDest: 354 Annotation: Subtype: Link Rect: 503, 271, 515, 280 ActionDest: 354 Annotation: Subtype: Link Rect: 310, 221, 322, 230 ActionDest: 354 Annotation: Subtype: Link Rect: 349, 180, 356, 192 ActionDest: 315 Annotation: Subtype: Link Rect: 346, 158, 358, 167 ActionDest: 354 Annotation: Subtype: Link Rect: 359, 158, 371, 167 ActionDest: 354 Page: Sequence: 16 Annotations: Annotation: Subtype: Link Rect: 438, 723, 450, 732 ActionDest: 354 Annotation: Subtype: Link Rect: 525, 634, 560, 645 Annotation: Subtype: Link Rect: 165, 622, 372, 633 Annotation: Subtype: Link Rect: 227, 137, 265, 148 Annotation: Subtype: Link Rect: 271, 137, 307, 148 Annotation: Subtype: Link Rect: 281, 103, 319, 114 Annotation: Subtype: Link Rect: 325, 103, 361, 114 Page: Sequence: 17 Annotations: Annotation: Subtype: Link Rect: 519, 710, 557, 721 Annotation: Subtype: Link Rect: 59, 699, 95, 710 Annotation: Subtype: Link Rect: 353, 665, 391, 675 Annotation: Subtype: Link Rect: 131, 642, 168, 652 Annotation: Subtype: Link Rect: 59, 607, 96, 617 Annotation: Subtype: Link Rect: 492, 583, 530, 594 Annotation: Subtype: Link Rect: 520, 572, 557, 583 Annotation: Subtype: Link Rect: 113, 550, 150, 560 Annotation: Subtype: Link Rect: 183, 526, 220, 537 Annotation: Subtype: Link Rect: 218, 503, 256, 514 Annotation: Subtype: Link Rect: 262, 503, 298, 514 Annotation: Subtype: Link Rect: 323, 480, 361, 491 Annotation: Subtype: Link Rect: 367, 480, 403, 491 Annotation: Subtype: Link Rect: 59, 446, 96, 456 Annotation: Subtype: Link Rect: 102, 446, 138, 456 Annotation: Subtype: Link Rect: 507, 422, 545, 433 Annotation: Subtype: Link Rect: 462, 399, 499, 410 Annotation: Subtype: Link Rect: 259, 377, 297, 387 Annotation: Subtype: Link Rect: 304, 353, 341, 364 Annotation: Subtype: Link Rect: 447, 330, 485, 341 Annotation: Subtype: Link Rect: 59, 297, 96, 307 Annotation: Subtype: Link Rect: 288, 273, 325, 284 Annotation: Subtype: Link Rect: 131, 251, 168, 261 Annotation: Subtype: Link Rect: 175, 251, 211, 261 Annotation: Subtype: Link Rect: 242, 216, 279, 226 Annotation: Subtype: Link Rect: 365, 192, 403, 203 Annotation: Subtype: Link Rect: 409, 192, 445, 203 Annotation: Subtype: Link Rect: 108, 159, 146, 169 Annotation: Subtype: Link Rect: 152, 159, 188, 169 Annotation: Subtype: Link Rect: 59, 124, 96, 134 Annotation: Subtype: Link Rect: 490, 112, 527, 123 Annotation: Subtype: Link Rect: 441, 89, 479, 100 Page: Sequence: 18 Annotations: Annotation: Subtype: Link Rect: 59, 711, 96, 721 Annotation: Subtype: Link Rect: 433, 687, 470, 698 Annotation: Subtype: Link Rect: 352, 664, 389, 675 Annotation: Subtype: Link Rect: 314, 641, 351, 652 Annotation: Subtype: Link Rect: 358, 641, 394, 652 Annotation: Subtype: Link Rect: 295, 606, 332, 617 Annotation: Subtype: Link Rect: 398, 583, 436, 594 Annotation: Subtype: Link Rect: 442, 583, 478, 594 Annotation: Subtype: Link Rect: 59, 550, 96, 560 Annotation: Subtype: Link Rect: 102, 550, 138, 560 Annotation: Subtype: Link Rect: 358, 526, 395, 537 Annotation: Subtype: Link Rect: 401, 526, 437, 537 Annotation: Subtype: Link Rect: 469, 503, 506, 514 Annotation: Subtype: Link Rect: 456, 480, 494, 491 Annotation: Subtype: Link Rect: 131, 458, 168, 468 Annotation: Subtype: Link Rect: 380, 434, 417, 445 Annotation: Subtype: Link Rect: 59, 400, 96, 410 Annotation: Subtype: Link Rect: 455, 376, 492, 387 Annotation: Subtype: Link Rect: 499, 376, 535, 387 Annotation: Subtype: Link Rect: 425, 353, 462, 364 Annotation: Subtype: Link Rect: 469, 353, 505, 364 Annotation: Subtype: Link Rect: 478, 330, 515, 341 Annotation: Subtype: Link Rect: 521, 330, 557, 341 Annotation: Subtype: Link Rect: 192, 307, 230, 318 Annotation: Subtype: Link Rect: 117, 274, 155, 284 Annotation: Subtype: Link Rect: 406, 250, 443, 261 Annotation: Subtype: Link Rect: 280, 227, 317, 238 Annotation: Subtype: Link Rect: 291, 192, 328, 203 Annotation: Subtype: Link Rect: 519, 169, 557, 180 Annotation: Subtype: Link Rect: 458, 146, 495, 157 Annotation: Subtype: Link Rect: 290, 123, 327, 134 Annotation: Subtype: Link Rect: 334, 123, 370, 134 Checksum: b58d5373 Type: CRC32 Checksum: 05e20cf6c1eaed21d047ac38748e5c20 Type: MD5 Checksum: 745d3d91bf55f3eadd0bb4ac00d1d09a313878ab Type: SHA-1