d4bede04c74290f34706fc213ce5dc3c NUT_a2022v14p3440.pdf 71b3e58167081ed113b759adee56d7c83fb7d1c5 NUT_a2022v14p3440.pdf 247a82d70c7187b13ed7c0d99353501b96cdcfe0d97130055337d8634da30f85 NUT_a2022v14p3440.pdf Title: A High-Fat Diet Modifies Brain Neurotransmitter Profile and Hippocampal Proteome and Morphology in an IUGR Pig Model Subject: Intrauterine Growth Restriction (IUGR) hinders the correct growth of the fetus during pregnancy due to the lack of oxygen or nutrients. The developing fetus gives priority to brain development (“brain sparing”), but the risk exists of neurological and cognitive deficits at short or long term. On the other hand, diets rich in fat exert pernicious effects on brain function. Using a pig model of spontaneous IUGR, we have studied the effect on the adult of a long-term high-fat diet (HFD) on the neurotransmitter profile in several brain areas, and the morphology and the proteome of the hippocampus. Our hypothesis was that animals affected by IUGR (born with low birth weight) would present a different susceptibility to an HFD when they become adults, compared with normal birth-weight animals. Our results indicate that HFD affected the serotoninergic pathway, but it did not provoke relevant changes in the morphology of the hippocampus. Finally, the proteomic analysis revealed that, in some instances, NBW and LBW individuals respond to HFD in different ways. In particular, NBW animals presented changes in oxidative phosphorylation and the extracellular matrix, whereas LBW animals presented differences in RNA splicing, anterograde and retrograde transport and the mTOR pathway. Keywords: neurotransmitters; metabolism; high-fat diet; hippocampus; intrauterine growth restriction; brain; pig Author: Natalia Yeste, Jorge Pérez-Valle, Ana Heras-Molina, José Luis Pesántez-Pacheco, Esteban Porrini, Antonio González-Bulnes and Anna Bassols Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Thu Aug 25 15:14:00 2022 CEST ModDate: Thu Aug 25 15:20:28 2022 CEST Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 26 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 3239673 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- TOZXSU+VnURWPalladioL Type 1 Custom yes yes no 10 0 DYRQZT+URWPalladioL-Bold Type 1 Custom yes yes no 15 0 CTASAJ+URWPalladioL-Roma Type 1 Custom yes yes no 20 0 JBTAZD+URWPalladioL-Ital Type 1 Custom yes yes no 24 0 GIFXFT+VnURWPalladioL-Bold Type 1 Custom yes yes no 28 0 HGMNRX+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 46 0 IFEDSM+CMSY10 Type 1 Builtin yes yes no 71 0 OVNEXW+EURM10 Type 1 Builtin yes yes no 75 0 WBVKDJ+URWPalladioL-BoldItal Type 1 Custom yes yes no 91 0 ABCDEE+Palatino Linotype,Bold TrueType WinAnsi yes yes no 106 0 ABCDEE+Palatino Linotype TrueType WinAnsi yes yes no 110 0 ABCDEE+Palatino Linotype,Italic TrueType WinAnsi yes yes no 114 0 ABCDEE+Palatino Linotype CID TrueType Identity-H yes yes yes 118 0 XIRVNA+EURB10 Type 1 Builtin yes yes no 131 0 MDIKPC+CMBSY10 Type 1 Builtin yes yes no 135 0 ABCDEE+Palatino Linotype,Bold TrueType WinAnsi yes yes no 142 0 ABCDEE+Palatino Linotype TrueType WinAnsi yes yes no 146 0 ABCDEE+Palatino Linotype,Italic TrueType WinAnsi yes yes no 150 0 ABCDEE+Palatino Linotype,Bold TrueType WinAnsi yes yes no 163 0 ABCDEE+Palatino Linotype TrueType WinAnsi yes yes no 167 0 ABCDEE+Palatino Linotype,Italic TrueType WinAnsi yes yes no 171 0 ABCDEE+Palatino Linotype,Bold TrueType WinAnsi yes yes no 184 0 ABCDEE+Palatino Linotype TrueType WinAnsi yes yes no 188 0 ABCDEE+Palatino Linotype,Italic TrueType WinAnsi yes yes no 192 0 ABCDEE+Palatino Linotype,Bold TrueType WinAnsi yes yes no 207 0 ABCDEE+Palatino Linotype TrueType WinAnsi yes yes no 211 0 ABCDEE+Palatino Linotype,Italic TrueType WinAnsi yes yes no 215 0 ABCDEE+Palatino Linotype,Bold TrueType WinAnsi yes yes no 230 0 ABCDEE+Palatino Linotype TrueType WinAnsi yes yes no 234 0 ABCDEE+Palatino Linotype,Italic TrueType WinAnsi yes yes no 238 0 ABCDEE+Palatino Linotype,Bold TrueType WinAnsi yes yes no 262 0 ABCDEE+Palatino Linotype TrueType WinAnsi yes yes no 266 0 ABCDEE+Palatino Linotype,Italic TrueType WinAnsi yes yes no 270 0 ABCDEE+Palatino Linotype CID TrueType Identity-H yes yes yes 274 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2022-11-29 02:32:21 CET RepresentationInformation: NUT_a2022v14p3440.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2022-11-28 13:49:14 CET Size: 3239673 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 552 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4923ab24 Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@44c8afef Children: Item: Title: Ethics Statement Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7b69c6ba Item: Title: Animals and Experimental Procedure Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@46daef40 Item: Title: Quantification of Neurotransmitters Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@12f41634 Item: Title: Immunohistochemical Analysis of the Hippocampus Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@13c27452 Item: Title: Image Processing and Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@262b2c86 Item: Title: Statistical Analyses Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@371a67ec Item: Title: Proteomic Analysis by Isobaric Mass Tag Labeling with TMT10plex™ Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@5ed828d Item: Title: Nanoliquid Chromatography Electrospray Ionization Tandem Mass Spectrometry (nanoLC-ESI-MS/MS) Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@50d0686 Item: Title: Database Searching Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7a3d45bd Item: Title: Quantitative Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1e7c7811 Item: Title: Gene Ontology and Bioinformatic Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77ec78b9 Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1a3869f4 Children: Item: Title: Effects of High-Fat Diet and the Influence of IUGR on the Neurotransmitter Profile in Several Brain Areas Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@a38d7a3 Item: Title: Effects of High-Fat Diet and the Influence of IUGR on the Morphology of the Hippocampus Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77f99a05 Item: Title: Effects of High-Fat Diet and the Influence of IUGR on the Proteome of the Hippocampus Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@63440df3 Children: Item: Title: Gene Ontology Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@3aeaafa6 Item: Title: KEGG Pathway Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@76a3e297 Item: Title: Pathway Analysis with Reactome Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4d3167f4 Item: Title: Upregulated and Downregulated Proteins Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@ed9d034 Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@6121c9d6 Children: Item: Title: Effect of an HFD Diet in NBW and LBW Pigs in the NT Profile and in the Morphology of the Hippocampus Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@87f383f Item: Title: Effect of an HFD Diet in NBW and LBW Pigs in the Proteome of the Hippocampus Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4eb7f003 Item: Title: Conclusions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@eafc191 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@612fc6eb Info: Title: A High-Fat Diet Modifies Brain Neurotransmitter Profile and Hippocampal Proteome and Morphology in an IUGR Pig Model Author: Natalia Yeste, Jorge Pérez-Valle, Ana Heras-Molina, José Luis Pesántez-Pacheco, Esteban Porrini, Antonio González-Bulnes and Anna Bassols Subject: Intrauterine Growth Restriction (IUGR) hinders the correct growth of the fetus during pregnancy due to the lack of oxygen or nutrients. The developing fetus gives priority to brain development (“brain sparing”), but the risk exists of neurological and cognitive deficits at short or long term. On the other hand, diets rich in fat exert pernicious effects on brain function. Using a pig model of spontaneous IUGR, we have studied the effect on the adult of a long-term high-fat diet (HFD) on the neurotransmitter profile in several brain areas, and the morphology and the proteome of the hippocampus. Our hypothesis was that animals affected by IUGR (born with low birth weight) would present a different susceptibility to an HFD when they become adults, compared with normal birth-weight animals. Our results indicate that HFD affected the serotoninergic pathway, but it did not provoke relevant changes in the morphology of the hippocampus. Finally, the proteomic analysis revealed that, in some instances, NBW and LBW individuals respond to HFD in different ways. In particular, NBW animals presented changes in oxidative phosphorylation and the extracellular matrix, whereas LBW animals presented differences in RNA splicing, anterograde and retrograde transport and the mTOR pathway. Keywords: neurotransmitters; metabolism; high-fat diet; hippocampus; intrauterine growth restriction; brain; pig Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Thu Aug 25 15:14:00 CEST 2022 ModDate: Thu Aug 25 15:20:28 CEST 2022 ID: 0xac0509c355cc065d2c2b6f4c6c91400a, 0xac0509c355cc065d2c2b6f4c6c91400a Filters: FilterPipeline: FlateDecode Fonts: Type1: Font: BaseFont: XIRVNA+EURB10 FontSubset: true FirstChar: 22 LastChar: 22 FontDescriptor: FontName: XIRVNA+EURB10 Flags: Symbolic FontBBox: -30, -247, 1178, 723 FontFile: true Font: BaseFont: CTASAJ+URWPalladioL-Roma FontSubset: true FirstChar: 2 LastChar: 252 FontDescriptor: FontName: CTASAJ+URWPalladioL-Roma Flags: Symbolic FontBBox: -166, -283, 1021, 943 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: IFEDSM+CMSY10 FontSubset: true FirstChar: 0 LastChar: 48 FontDescriptor: FontName: IFEDSM+CMSY10 Flags: Symbolic FontBBox: -29, -960, 1116, 775 FontFile: true Font: BaseFont: MDIKPC+CMBSY10 FontSubset: true FirstChar: 6 LastChar: 6 FontDescriptor: FontName: MDIKPC+CMBSY10 Flags: Symbolic FontBBox: -26, -940, 1332, 825 FontFile: true Font: BaseFont: JBTAZD+URWPalladioL-Ital FontSubset: true FirstChar: 2 LastChar: 153 FontDescriptor: FontName: JBTAZD+URWPalladioL-Ital Flags: Symbolic FontBBox: -170, -305, 1010, 941 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: TOZXSU+VnURWPalladioL FontSubset: true FirstChar: 160 LastChar: 226 FontDescriptor: FontName: TOZXSU+VnURWPalladioL Flags: Symbolic FontBBox: -166, -283, 1021, 943 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: OVNEXW+EURM10 FontSubset: true FirstChar: 22 LastChar: 22 FontDescriptor: FontName: OVNEXW+EURM10 Flags: Symbolic FontBBox: -32, -243, 1060, 720 FontFile: true Font: BaseFont: WBVKDJ+URWPalladioL-BoldItal FontSubset: true FirstChar: 112 LastChar: 112 FontDescriptor: FontName: WBVKDJ+URWPalladioL-BoldItal Flags: Symbolic FontBBox: -170, -300, 1073, 935 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: GIFXFT+VnURWPalladioL-Bold FontSubset: true FirstChar: 161 LastChar: 178 FontDescriptor: FontName: GIFXFT+VnURWPalladioL-Bold Flags: Symbolic FontBBox: -152, -301, 1000, 935 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: DYRQZT+URWPalladioL-Bold FontSubset: true FirstChar: 2 LastChar: 122 FontDescriptor: FontName: DYRQZT+URWPalladioL-Bold Flags: Symbolic FontBBox: -152, -301, 1000, 935 FontFile: true EncodingDictionary: Differences: true Pages: Page: Sequence: 1 Annotations: Annotation: Subtype: Link Rect: 112, 347, 152, 356 Annotation: Subtype: Link Rect: 34, 335, 126, 344 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, 757, 169, 793 Annotation: Subtype: Link Rect: 515, 757, 560, 793 Annotation: Subtype: Link Rect: 100, 661, 111, 675 Annotation: Subtype: Link Rect: 295, 661, 306, 675 Annotation: Subtype: Link Rect: 439, 661, 450, 675 Annotation: Subtype: Link Rect: 159, 651, 170, 662 Annotation: Subtype: Link Rect: 265, 651, 276, 662 Annotation: Subtype: Link Rect: 342, 206, 349, 215 ActionDest: 286 Annotation: Subtype: Link Rect: 440, 193, 447, 202 ActionDest: 286 Annotation: Subtype: Link Rect: 448, 193, 455, 202 ActionDest: 286 Annotation: Subtype: Link Rect: 281, 168, 288, 177 ActionDest: 286 Annotation: Subtype: Link Rect: 291, 168, 298, 177 ActionDest: 286 Annotation: Subtype: Link Rect: 295, 143, 302, 152 ActionDest: 286 Annotation: Subtype: Link Rect: 198, 118, 210, 127 ActionDest: 286 Annotation: Subtype: Link Rect: 211, 118, 223, 127 ActionDest: 286 Annotation: Subtype: Link Rect: 120, 17, 254, 28 Annotation: Subtype: Link Rect: 404, 17, 560, 28 Annotation: Subtype: Link Rect: 34, 444, 87, 463 Page: Sequence: 2 Annotations: Annotation: Subtype: Link Rect: 207, 660, 219, 669 ActionDest: 286 Annotation: Subtype: Link Rect: 375, 635, 387, 644 ActionDest: 286 Annotation: Subtype: Link Rect: 390, 635, 402, 644 ActionDest: 286 Annotation: Subtype: Link Rect: 254, 610, 266, 619 ActionDest: 289 Annotation: Subtype: Link Rect: 269, 610, 281, 619 ActionDest: 289 Annotation: Subtype: Link Rect: 444, 585, 456, 594 ActionDest: 289 Annotation: Subtype: Link Rect: 457, 585, 469, 594 ActionDest: 289 Annotation: Subtype: Link Rect: 239, 497, 251, 506 ActionDest: 289 Annotation: Subtype: Link Rect: 252, 497, 264, 506 ActionDest: 289 Annotation: Subtype: Link Rect: 185, 459, 197, 468 ActionDest: 289 Annotation: Subtype: Link Rect: 200, 459, 212, 468 ActionDest: 289 Annotation: Subtype: Link Rect: 518, 447, 530, 456 ActionDest: 289 Annotation: Subtype: Link Rect: 531, 447, 543, 456 ActionDest: 289 Annotation: Subtype: Link Rect: 316, 434, 328, 443 ActionDest: 289 Annotation: Subtype: Link Rect: 350, 422, 362, 431 ActionDest: 289 Annotation: Subtype: Link Rect: 365, 422, 377, 431 ActionDest: 289 Annotation: Subtype: Link Rect: 195, 346, 207, 355 ActionDest: 289 Annotation: Subtype: Link Rect: 208, 346, 220, 355 ActionDest: 289 Annotation: Subtype: Link Rect: 195, 321, 207, 330 ActionDest: 289 Annotation: Subtype: Link Rect: 210, 321, 222, 330 ActionDest: 289 Page: Sequence: 3 Annotations: Annotation: Subtype: Link Rect: 192, 205, 204, 214 ActionDest: 289 Page: Sequence: 4 Page: Sequence: 5 Page: Sequence: 6 Annotations: Annotation: Subtype: Link Rect: 436, 506, 448, 516 ActionDest: 292 Annotation: Subtype: Link Rect: 209, 479, 324, 491 Annotation: Subtype: Link Rect: 386, 391, 465, 403 Annotation: Subtype: Link Rect: 232, 365, 338, 377 Annotation: Subtype: Link Rect: 352, 353, 469, 365 Annotation: Subtype: Link Rect: 168, 290, 272, 302 Annotation: Subtype: Link Rect: 396, 293, 408, 302 ActionDest: 292 Annotation: Subtype: Link Rect: 218, 278, 422, 290 Annotation: Subtype: Link Rect: 544, 280, 556, 289 ActionDest: 292 Annotation: Subtype: Link Rect: 223, 252, 322, 264 Annotation: Subtype: Link Rect: 443, 255, 455, 264 ActionDest: 292 Annotation: Subtype: Link Rect: 300, 230, 312, 239 ActionDest: 292 Annotation: Subtype: Link Rect: 310, 165, 317, 177 ActionDest: 95 Page: Sequence: 7 Page: Sequence: 8 Page: Sequence: 9 Annotations: Annotation: Subtype: Link Rect: 299, 717, 306, 729 ActionDest: 127 Annotation: Subtype: Link Rect: 357, 717, 364, 729 ActionDest: 98 Annotation: Subtype: Link Rect: 503, 190, 510, 202 ActionDest: 127 Page: Sequence: 10 Page: Sequence: 11 Annotations: Annotation: Subtype: Link Rect: 541, 620, 548, 632 ActionDest: 156 Annotation: Subtype: Link Rect: 441, 582, 448, 594 ActionDest: 156 Annotation: Subtype: Link Rect: 199, 129, 206, 141 ActionDest: 177 Page: Sequence: 12 Annotations: Annotation: Subtype: Link Rect: 345, 277, 352, 289 ActionDest: 200 Annotation: Subtype: Link Rect: 446, 214, 453, 227 ActionDest: 197 Page: Sequence: 13 Page: Sequence: 14 Annotations: Annotation: Subtype: Link Rect: 297, 399, 304, 411 ActionDest: 220 Annotation: Subtype: Link Rect: 455, 248, 462, 260 ActionDest: 223 Page: Sequence: 15 Page: Sequence: 16 Annotations: Annotation: Subtype: Link Rect: 237, 170, 244, 182 ActionDest: 243 Page: Sequence: 17 Page: Sequence: 18 Annotations: Annotation: Subtype: Link Rect: 270, 657, 277, 666 ActionDest: 286 Annotation: Subtype: Link Rect: 278, 657, 290, 666 ActionDest: 286 Annotation: Subtype: Link Rect: 286, 582, 298, 591 ActionDest: 292 Annotation: Subtype: Link Rect: 299, 582, 311, 591 ActionDest: 292 Annotation: Subtype: Link Rect: 240, 557, 252, 566 ActionDest: 289 Annotation: Subtype: Link Rect: 253, 557, 265, 566 ActionDest: 289 Annotation: Subtype: Link Rect: 265, 557, 277, 566 ActionDest: 292 Annotation: Subtype: Link Rect: 280, 557, 292, 566 ActionDest: 292 Annotation: Subtype: Link Rect: 363, 532, 375, 541 ActionDest: 289 Annotation: Subtype: Link Rect: 375, 532, 387, 541 ActionDest: 289 Annotation: Subtype: Link Rect: 223, 332, 235, 342 ActionDest: 292 Annotation: Subtype: Link Rect: 235, 332, 247, 342 ActionDest: 292 Annotation: Subtype: Link Rect: 193, 270, 205, 279 ActionDest: 292 Annotation: Subtype: Link Rect: 205, 270, 217, 279 ActionDest: 292 Annotation: Subtype: Link Rect: 248, 245, 260, 254 ActionDest: 292 Annotation: Subtype: Link Rect: 261, 245, 273, 254 ActionDest: 292 Annotation: Subtype: Link Rect: 543, 144, 555, 153 ActionDest: 289 Annotation: Subtype: Link Rect: 447, 69, 459, 78 ActionDest: 292 Page: Sequence: 19 Annotations: Annotation: Subtype: Link Rect: 517, 723, 529, 732 ActionDest: 292 Annotation: Subtype: Link Rect: 272, 698, 284, 707 ActionDest: 289 Annotation: Subtype: Link Rect: 419, 484, 431, 493 ActionDest: 289 Annotation: Subtype: Link Rect: 302, 381, 309, 393 ActionDest: 177 Annotation: Subtype: Link Rect: 415, 258, 427, 267 ActionDest: 289 Annotation: Subtype: Link Rect: 195, 105, 202, 117 ActionDest: 197 Page: Sequence: 20 Annotations: Annotation: Subtype: Link Rect: 197, 622, 209, 631 ActionDest: 292 Annotation: Subtype: Link Rect: 212, 622, 224, 631 ActionDest: 292 Annotation: Subtype: Link Rect: 439, 597, 451, 606 ActionDest: 292 Annotation: Subtype: Link Rect: 441, 572, 453, 581 ActionDest: 292 Annotation: Subtype: Link Rect: 253, 535, 265, 544 ActionDest: 292 Annotation: Subtype: Link Rect: 323, 422, 335, 431 ActionDest: 289 Annotation: Subtype: Link Rect: 467, 271, 479, 280 ActionDest: 292 Annotation: Subtype: Link Rect: 482, 271, 494, 280 ActionDest: 295 Annotation: Subtype: Link Rect: 490, 208, 502, 217 ActionDest: 295 Annotation: Subtype: Link Rect: 226, 170, 238, 180 ActionDest: 295 Annotation: Subtype: Link Rect: 388, 145, 400, 154 ActionDest: 295 Annotation: Subtype: Link Rect: 437, 95, 449, 104 ActionDest: 292 Annotation: Subtype: Link Rect: 450, 95, 462, 104 ActionDest: 292 Page: Sequence: 21 Annotations: Annotation: Subtype: Link Rect: 260, 698, 272, 707 ActionDest: 295 Annotation: Subtype: Link Rect: 273, 698, 285, 707 ActionDest: 295 Annotation: Subtype: Link Rect: 307, 570, 314, 582 ActionDest: 255 Annotation: Subtype: Link Rect: 472, 535, 484, 544 ActionDest: 295 Annotation: Subtype: Link Rect: 484, 535, 496, 544 ActionDest: 295 Page: Sequence: 22 Annotations: Annotation: Subtype: Link Rect: 239, 736, 251, 744 ActionDest: 295 Annotation: Subtype: Link Rect: 251, 735, 263, 744 ActionDest: 295 Annotation: Subtype: Link Rect: 199, 698, 211, 707 ActionDest: 295 Annotation: Subtype: Link Rect: 278, 698, 290, 707 ActionDest: 295 Annotation: Subtype: Link Rect: 529, 597, 541, 606 ActionDest: 292 Annotation: Subtype: Link Rect: 544, 597, 556, 606 ActionDest: 292 Annotation: Subtype: Link Rect: 328, 572, 340, 581 ActionDest: 289 Annotation: Subtype: Link Rect: 341, 572, 353, 581 ActionDest: 295 Annotation: Subtype: Link Rect: 261, 560, 273, 569 ActionDest: 289 Annotation: Subtype: Link Rect: 536, 295, 560, 306 Annotation: Subtype: Link Rect: 165, 282, 369, 293 Page: Sequence: 23 Annotations: Annotation: Subtype: Link Rect: 365, 418, 402, 429 Annotation: Subtype: Link Rect: 409, 418, 445, 429 Annotation: Subtype: Link Rect: 372, 395, 409, 406 Annotation: Subtype: Link Rect: 415, 395, 452, 406 Annotation: Subtype: Link Rect: 518, 372, 556, 383 Annotation: Subtype: Link Rect: 95, 350, 132, 360 Annotation: Subtype: Link Rect: 104, 327, 141, 337 Annotation: Subtype: Link Rect: 95, 304, 133, 314 Annotation: Subtype: Link Rect: 139, 304, 175, 314 Annotation: Subtype: Link Rect: 479, 292, 516, 303 Annotation: Subtype: Link Rect: 158, 270, 196, 280 Annotation: Subtype: Link Rect: 440, 257, 477, 268 Annotation: Subtype: Link Rect: 95, 235, 132, 245 Annotation: Subtype: Link Rect: 519, 211, 557, 222 Annotation: Subtype: Link Rect: 59, 201, 95, 211 Annotation: Subtype: Link Rect: 357, 177, 395, 188 Annotation: Subtype: Link Rect: 216, 154, 254, 165 Annotation: Subtype: Link Rect: 260, 154, 296, 165 Annotation: Subtype: Link Rect: 227, 131, 265, 142 Annotation: Subtype: Link Rect: 124, 97, 161, 107 Annotation: Subtype: Link Rect: 171, 62, 208, 73 Page: Sequence: 24 Annotations: Annotation: Subtype: Link Rect: 494, 722, 531, 733 Annotation: Subtype: Link Rect: 339, 688, 377, 698 Annotation: Subtype: Link Rect: 463, 664, 501, 675 Annotation: Subtype: Link Rect: 516, 652, 553, 664 Annotation: Subtype: Link Rect: 312, 629, 349, 640 Annotation: Subtype: Link Rect: 355, 629, 391, 640 Annotation: Subtype: Link Rect: 184, 596, 221, 606 Annotation: Subtype: Link Rect: 228, 596, 264, 606 Annotation: Subtype: Link Rect: 181, 549, 219, 560 Annotation: Subtype: Link Rect: 225, 549, 261, 560 Annotation: Subtype: Link Rect: 430, 514, 467, 525 Annotation: Subtype: Link Rect: 178, 481, 216, 491 Annotation: Subtype: Link Rect: 229, 445, 267, 456 Annotation: Subtype: Link Rect: 59, 412, 96, 422 Annotation: Subtype: Link Rect: 159, 377, 197, 387 Annotation: Subtype: Link Rect: 425, 342, 463, 353 Annotation: Subtype: Link Rect: 304, 307, 342, 318 Annotation: Subtype: Link Rect: 226, 274, 263, 284 Annotation: Subtype: Link Rect: 81, 239, 119, 249 Annotation: Subtype: Link Rect: 125, 239, 161, 249 Annotation: Subtype: Link Rect: 95, 216, 132, 226 Annotation: Subtype: Link Rect: 138, 216, 174, 226 Annotation: Subtype: Link Rect: 59, 182, 96, 192 Annotation: Subtype: Link Rect: 102, 182, 138, 192 Annotation: Subtype: Link Rect: 467, 158, 505, 169 Annotation: Subtype: Link Rect: 59, 124, 96, 134 Annotation: Subtype: Link Rect: 504, 100, 542, 111 Annotation: Subtype: Link Rect: 422, 77, 460, 88 Page: Sequence: 25 Annotations: Annotation: Subtype: Link Rect: 353, 722, 391, 733 Annotation: Subtype: Link Rect: 379, 699, 417, 710 Annotation: Subtype: Link Rect: 59, 676, 96, 687 Annotation: Subtype: Link Rect: 102, 676, 138, 687 Annotation: Subtype: Link Rect: 369, 641, 407, 652 Annotation: Subtype: Link Rect: 413, 641, 449, 652 Annotation: Subtype: Link Rect: 216, 607, 253, 617 Annotation: Subtype: Link Rect: 259, 607, 295, 617 Annotation: Subtype: Link Rect: 281, 584, 319, 594 Annotation: Subtype: Link Rect: 325, 584, 361, 594 Annotation: Subtype: Link Rect: 59, 550, 96, 560 Annotation: Subtype: Link Rect: 86, 515, 124, 525 Annotation: Subtype: Link Rect: 370, 491, 407, 502 Annotation: Subtype: Link Rect: 297, 468, 334, 479 Annotation: Subtype: Link Rect: 382, 445, 419, 456 Annotation: Subtype: Link Rect: 282, 422, 320, 433 Annotation: Subtype: Link Rect: 113, 400, 150, 410 Annotation: Subtype: Link Rect: 157, 400, 193, 410 Annotation: Subtype: Link Rect: 284, 376, 321, 387 Annotation: Subtype: Link Rect: 328, 376, 364, 387 Annotation: Subtype: Link Rect: 279, 354, 316, 364 Annotation: Subtype: Link Rect: 332, 330, 370, 341 Annotation: Subtype: Link Rect: 59, 308, 96, 318 Annotation: Subtype: Link Rect: 131, 285, 168, 295 Annotation: Subtype: Link Rect: 352, 261, 390, 272 Annotation: Subtype: Link Rect: 157, 239, 195, 249 Annotation: Subtype: Link Rect: 350, 204, 387, 215 Annotation: Subtype: Link Rect: 154, 170, 191, 180 Annotation: Subtype: Link Rect: 431, 146, 468, 157 Annotation: Subtype: Link Rect: 475, 146, 511, 157 Annotation: Subtype: Link Rect: 298, 123, 336, 134 Annotation: Subtype: Link Rect: 342, 123, 378, 134 Annotation: Subtype: Link Rect: 413, 100, 451, 111 Annotation: Subtype: Link Rect: 457, 100, 493, 111 Annotation: Subtype: Link Rect: 153, 78, 191, 88 Annotation: Subtype: Link Rect: 424, 66, 461, 77 Page: Sequence: 26 Annotations: Annotation: Subtype: Link Rect: 247, 722, 284, 733 Annotation: Subtype: Link Rect: 379, 699, 417, 710 Annotation: Subtype: Link Rect: 296, 676, 333, 687 Annotation: Subtype: Link Rect: 311, 641, 349, 652 Annotation: Subtype: Link Rect: 209, 618, 247, 629 Annotation: Subtype: Link Rect: 209, 596, 246, 606 Annotation: Subtype: Link Rect: 253, 596, 289, 606 Annotation: Subtype: Link Rect: 135, 573, 173, 583 Annotation: Subtype: Link Rect: 179, 573, 215, 583 Annotation: Subtype: Link Rect: 188, 550, 225, 560 Annotation: Subtype: Link Rect: 231, 550, 267, 560 Annotation: Subtype: Link Rect: 95, 527, 133, 537 Annotation: Subtype: Link Rect: 187, 503, 224, 514 Annotation: Subtype: Link Rect: 329, 481, 367, 491 Annotation: Subtype: Link Rect: 146, 458, 183, 468 Annotation: Subtype: Link Rect: 297, 422, 335, 433 Checksum: 1b1f2b8a Type: CRC32 Checksum: d4bede04c74290f34706fc213ce5dc3c Type: MD5 Checksum: 71b3e58167081ed113b759adee56d7c83fb7d1c5 Type: SHA-1