4c2936024c1533f1adf888795b01125e pmc_10459912.pdf da8c88f1448cf98746f8c89018a47abbc399973a pmc_10459912.pdf c2b96459b8724dc490779a1307bc9bca024fce4fe97ecfcabd1b14b41fca0d5d pmc_10459912.pdf Title: Dietary 3 Fatty Acids and Phytosterols in the Modulation of the HDL Lipidome: A Longitudinal Crossover Clinical Study Subject: High-density lipoproteins (HDLs) are complex particles composed of a wide range of lipids, proteins, hormones and vitamins that confer to the HDL particles multiple cardiovascular protective properties, mainly against the development of atherosclerosis. Among other factors, the HDL lipidome is affected by diet. We hypothesized that diet supplementation with 3 (docosahexaenoic acid: DHA and eicosapentaenoic acid: EPA) and phytosterols (PhyS) would improve the HDL lipid profile. Overweight subjects (n = 20) were enrolled in a two-arm longitudinal crossover study. Milk (250 mL/day), supplemented with either 3 (EPA + DHA, 375 mg) or PhyS (1.6 g), was administered to the volunteers over two consecutive 28-day intervention periods, followed by HDL lipidomic analysis. The comprehensive lipid pattern revealed that the HDL lipidome is diet-dependent. 3-milk supplementation produced more changes than PhyS, mainly in cholesteryl esters (CEs). After 3-milk intake, levels of DHA and EPA within phosphatylcholines, triglycerides and CE lipids in HDLs increased (p < 0.05). The correlation between lipid species showed that lipid changes occur in a coordinated manner. Finally, our analysis revealed that the HDL lipidome is also sex-dependent. The HDL lipidome is affected by diet and sex, and the 4 weeks of 3 supplementation induced HDL enrichment with EPA and DHA. Keywords: lipids; high-density lipoproteins; DHA; EPA; omega-3 fatty acids; sex Author: Teresa Padro, Anallely López-Yerena, Antonio Pérez, Gemma Vilahur and Lina Badimon Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Mon Aug 21 07:10:54 2023 CEST ModDate: Mon Aug 21 07:15:28 2023 CEST Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 17 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 2671002 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- NYWBCD+EURB10 Type 1 Builtin yes yes no 10 0 HJOMHV+VnURWPalladioL-Bold Type 1 Custom yes yes no 14 0 YRSWDI+VnURWPalladioL Type 1 Custom yes yes no 19 0 PMTDSU+EURM10 Type 1 Builtin yes yes no 23 0 HCOIOZ+URWPalladioL-Bold Type 1 Custom yes yes no 27 0 XQYHRL+URWPalladioL-Roma Type 1 Custom yes yes no 32 0 VDPHAI+URWPalladioL-Ital Type 1 Custom yes yes no 36 0 DDZECJ+EURM7 Type 1 Builtin yes yes no 40 0 HGMNRX+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 58 0 QMDDXF+CMSY10 Type 1 Builtin yes yes no 83 0 DODINH+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 94 0 DODINI+PalatinoLinotype CID TrueType Identity-H yes yes yes 97 0 DODINJ+PalatinoLinotype TrueType WinAnsi yes yes no 103 0 DODIOK+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 106 0 DODKCE+PalatinoLinotype TrueType MacRoman yes yes no 109 0 PDXFNG+PazoMath-Bold Type 1 Builtin yes yes no 119 0 WBVKDJ+URWPalladioL-BoldItal Type 1 Custom yes yes no 123 0 DODINH+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 137 0 DODINI+PalatinoLinotype CID TrueType Identity-H yes yes yes 140 0 DODINJ+PalatinoLinotype TrueType WinAnsi yes yes no 146 0 DODIOK+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 149 0 DODKCE+PalatinoLinotype TrueType MacRoman yes yes no 152 0 DODMOG+PalatinoLinotype,Italic TrueType MacRoman yes yes no 169 0 DODINH+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 172 0 DODINI+PalatinoLinotype CID TrueType Identity-H yes yes yes 175 0 DODINJ+PalatinoLinotype TrueType WinAnsi yes yes no 181 0 DODINK+PalatinoLinotype,Italic CID TrueType Identity-H yes yes yes 184 0 DODIOK+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 190 0 DODKCE+PalatinoLinotype TrueType MacRoman yes yes no 193 0 DODINH+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 210 0 DODINI+PalatinoLinotype CID TrueType Identity-H yes yes yes 213 0 DODINJ+PalatinoLinotype TrueType WinAnsi yes yes no 219 0 DODIOK+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 222 0 DODKCE+PalatinoLinotype TrueType MacRoman yes yes no 225 0 DODINH+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 240 0 DODINI+PalatinoLinotype CID TrueType Identity-H yes yes yes 243 0 DODINJ+PalatinoLinotype TrueType WinAnsi yes yes no 249 0 DODIOK+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 252 0 DODKCE+PalatinoLinotype TrueType MacRoman yes yes no 255 0 DODINH+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 269 0 DODINI+PalatinoLinotype CID TrueType Identity-H yes yes yes 272 0 DODINJ+PalatinoLinotype TrueType WinAnsi yes yes no 278 0 DODIOK+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 281 0 DODKCE+PalatinoLinotype TrueType MacRoman yes yes no 284 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-05-07 02:39:01 CEST RepresentationInformation: pmc_10459912.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2024-04-24 02:39:53 CEST Size: 2671002 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 476 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Materials and Methods Destination: section.2 Children: Item: Title: Study Design Destination: subsection.2.1 Item: Title: Blood Sampling Destination: subsection.2.2 Item: Title: HDL Sample Preparation and Purity Control Destination: subsection.2.3 Item: Title: Metabolite Extraction Destination: subsection.2.4 Item: Title: Lipidomic Pattern Analysis Destination: subsection.2.5 Item: Title: Statistics Destination: subsection.2.6 Item: Title: Results Destination: section.3 Children: Item: Title: HDL Lipidome Destination: subsection.3.1 Item: Title: Effects of Dietary Treatments on HDL Lipid Profile Destination: subsection.3.2 Children: Item: Title: Effect of 3 Milk on the HDL Lipidome Destination: subsubsection.3.2.1 Item: Title: Effect of PhyS Milk on the HDL Lipidome Destination: subsubsection.3.2.2 Item: Title: Sex-Related Differences in the Effects of Milk Supplementation Destination: subsection.3.3 Item: Title: Discussion Destination: section.4 Item: Title: Conclusions Destination: section.5 Item: Title: References Destination: section.A. Info: Title: Dietary 3 Fatty Acids and Phytosterols in the Modulation of the HDL Lipidome: A Longitudinal Crossover Clinical Study Author: Teresa Padro, Anallely López-Yerena, Antonio Pérez, Gemma Vilahur and Lina Badimon Subject: High-density lipoproteins (HDLs) are complex particles composed of a wide range of lipids, proteins, hormones and vitamins that confer to the HDL particles multiple cardiovascular protective properties, mainly against the development of atherosclerosis. Among other factors, the HDL lipidome is affected by diet. We hypothesized that diet supplementation with 3 (docosahexaenoic acid: DHA and eicosapentaenoic acid: EPA) and phytosterols (PhyS) would improve the HDL lipid profile. Overweight subjects (n = 20) were enrolled in a two-arm longitudinal crossover study. Milk (250 mL/day), supplemented with either 3 (EPA + DHA, 375 mg) or PhyS (1.6 g), was administered to the volunteers over two consecutive 28-day intervention periods, followed by HDL lipidomic analysis. The comprehensive lipid pattern revealed that the HDL lipidome is diet-dependent. 3-milk supplementation produced more changes than PhyS, mainly in cholesteryl esters (CEs). After 3-milk intake, levels of DHA and EPA within phosphatylcholines, triglycerides and CE lipids in HDLs increased (p < 0.05). The correlation between lipid species showed that lipid changes occur in a coordinated manner. Finally, our analysis revealed that the HDL lipidome is also sex-dependent. The HDL lipidome is affected by diet and sex, and the 4 weeks of 3 supplementation induced HDL enrichment with EPA and DHA. Keywords: lipids; high-density lipoproteins; DHA; EPA; omega-3 fatty acids; sex Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Mon Aug 21 07:10:54 CEST 2023 ModDate: Mon Aug 21 07:15:28 CEST 2023 ID: 0xb378f77cb906d87a01b4307e9d7e4f16, 0xb378f77cb906d87a01b4307e9d7e4f16 Filters: FilterPipeline: FlateDecode Fonts: Type1: Font: BaseFont: XQYHRL+URWPalladioL-Roma FontSubset: true FirstChar: 2 LastChar: 246 FontDescriptor: FontName: XQYHRL+URWPalladioL-Roma Flags: Symbolic FontBBox: -166, -283, 1021, 943 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: YRSWDI+VnURWPalladioL FontSubset: true FirstChar: 160 LastChar: 243 FontDescriptor: FontName: YRSWDI+VnURWPalladioL Flags: Symbolic FontBBox: -166, -283, 1021, 943 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: QMDDXF+CMSY10 FontSubset: true FirstChar: 0 LastChar: 14 FontDescriptor: FontName: QMDDXF+CMSY10 Flags: Symbolic FontBBox: -29, -960, 1116, 775 FontFile: true Font: BaseFont: VDPHAI+URWPalladioL-Ital FontSubset: true FirstChar: 2 LastChar: 121 FontDescriptor: FontName: VDPHAI+URWPalladioL-Ital Flags: Symbolic FontBBox: -170, -305, 1010, 941 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: PMTDSU+EURM10 FontSubset: true FirstChar: 12 LastChar: 33 FontDescriptor: FontName: PMTDSU+EURM10 Flags: Symbolic FontBBox: -32, -243, 1060, 720 FontFile: true Font: BaseFont: PDXFNG+PazoMath-Bold FontSubset: true FirstChar: 68 LastChar: 68 FontDescriptor: FontName: PDXFNG+PazoMath-Bold Flags: Symbolic FontBBox: -10, -266, 829, 695 FontFile: true Font: BaseFont: DDZECJ+EURM7 FontSubset: true FirstChar: 33 LastChar: 33 FontDescriptor: FontName: DDZECJ+EURM7 Flags: Symbolic FontBBox: -5, -243, 1229, 720 FontFile: true Font: BaseFont: NYWBCD+EURB10 FontSubset: true FirstChar: 33 LastChar: 33 FontDescriptor: FontName: NYWBCD+EURB10 Flags: Symbolic FontBBox: -30, -247, 1178, 723 FontFile: true Font: BaseFont: HCOIOZ+URWPalladioL-Bold FontSubset: true FirstChar: 3 LastChar: 134 FontDescriptor: FontName: HCOIOZ+URWPalladioL-Bold Flags: Symbolic FontBBox: -152, -301, 1000, 935 FontFile: true EncodingDictionary: Differences: 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: HJOMHV+VnURWPalladioL-Bold FontSubset: true FirstChar: 178 LastChar: 226 FontDescriptor: FontName: HJOMHV+VnURWPalladioL-Bold Flags: Symbolic FontBBox: -152, -301, 1000, 935 FontFile: true EncodingDictionary: Differences: true Pages: Page: Sequence: 1 Annotations: Annotation: Subtype: Link Rect: 82, 293, 152, 302 Annotation: Subtype: Link Rect: 34, 282, 99, 290 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, 169, 792 Annotation: Subtype: Link Rect: 515, 756, 560, 792 Annotation: Subtype: Link Rect: 119, 660, 130, 674 Annotation: Subtype: Link Rect: 250, 660, 261, 674 Annotation: Subtype: Link Rect: 342, 660, 353, 674 Annotation: Subtype: Link Rect: 549, 660, 560, 674 Annotation: Subtype: Link Rect: 397, 200, 404, 209 ActionDest: 298 Annotation: Subtype: Link Rect: 210, 162, 217, 171 ActionDest: 298 Annotation: Subtype: Link Rect: 482, 137, 489, 146 ActionDest: 298 Annotation: Subtype: Link Rect: 295, 87, 302, 96 ActionDest: 298 Annotation: Subtype: Link Rect: 120, 17, 254, 28 Annotation: Subtype: Link Rect: 404, 17, 560, 28 Annotation: Subtype: Link Rect: 34, 378, 87, 397 Page: Sequence: 2 Annotations: Annotation: Subtype: Link Rect: 527, 735, 534, 744 ActionDest: 298 Annotation: Subtype: Link Rect: 357, 698, 364, 707 ActionDest: 298 Annotation: Subtype: Link Rect: 285, 660, 292, 669 ActionDest: 298 Annotation: Subtype: Link Rect: 476, 635, 483, 644 ActionDest: 298 Annotation: Subtype: Link Rect: 207, 622, 214, 631 ActionDest: 298 Annotation: Subtype: Link Rect: 276, 622, 283, 631 ActionDest: 298 Annotation: Subtype: Link Rect: 544, 572, 556, 581 ActionDest: 298 Annotation: Subtype: Link Rect: 369, 547, 381, 556 ActionDest: 298 Annotation: Subtype: Link Rect: 398, 497, 410, 506 ActionDest: 298 Annotation: Subtype: Link Rect: 335, 459, 347, 468 ActionDest: 298 Annotation: Subtype: Link Rect: 269, 409, 281, 418 ActionDest: 301 Annotation: Subtype: Link Rect: 526, 371, 538, 380 ActionDest: 301 Annotation: Subtype: Link Rect: 334, 334, 346, 343 ActionDest: 301 Annotation: Subtype: Link Rect: 490, 232, 502, 241 ActionDest: 301 Annotation: Subtype: Link Rect: 432, 192, 439, 204 ActionDest: 80 Annotation: Subtype: Link Rect: 502, 182, 513, 191 ActionDest: 301 Page: Sequence: 3 Annotations: Annotation: Subtype: Link Rect: 402, 698, 414, 707 ActionDest: 301 Annotation: Subtype: Link Rect: 221, 119, 233, 128 ActionDest: 301 Annotation: Subtype: Link Rect: 234, 119, 246, 128 ActionDest: 301 Annotation: Subtype: Link Rect: 286, 116, 293, 128 ActionDest: 80 Page: Sequence: 4 Annotations: Annotation: Subtype: Link Rect: 451, 720, 463, 729 ActionDest: 301 Annotation: Subtype: Link Rect: 448, 544, 459, 553 ActionDest: 301 Annotation: Subtype: Link Rect: 460, 544, 472, 553 ActionDest: 301 Annotation: Subtype: Link Rect: 302, 92, 314, 101 ActionDest: 301 Annotation: Subtype: Link Rect: 303, 67, 315, 76 ActionDest: 301 Page: Sequence: 5 Annotations: Annotation: Subtype: Link Rect: 427, 720, 559, 732 Annotation: Subtype: Link Rect: 388, 560, 400, 568 ActionDest: 301 Annotation: Subtype: Link Rect: 165, 532, 397, 544 Annotation: Subtype: Link Rect: 401, 534, 413, 543 ActionDest: 301 Annotation: Subtype: Link Rect: 202, 396, 209, 405 ActionDest: 127 Page: Sequence: 6 Annotations: Annotation: Subtype: Link Rect: 495, 485, 502, 497 ActionDest: 127 Annotation: Subtype: Link Rect: 239, 460, 246, 472 ActionDest: 127 Page: Sequence: 7 Annotations: Annotation: Subtype: Link Rect: 545, 585, 552, 597 ActionDest: 127 Annotation: Subtype: Link Rect: 461, 523, 468, 535 ActionDest: 127 Annotation: Subtype: Link Rect: 285, 447, 292, 459 ActionDest: 159 Annotation: Subtype: Link Rect: 199, 397, 206, 409 ActionDest: 159 Annotation: Subtype: Link Rect: 193, 333, 200, 345 ActionDest: 198 Annotation: Subtype: Link Rect: 364, 245, 371, 257 ActionDest: 198 Annotation: Subtype: Link Rect: 218, 157, 225, 169 ActionDest: 230 Page: Sequence: 8 Page: Sequence: 9 Page: Sequence: 10 Annotations: Annotation: Subtype: Link Rect: 378, 274, 390, 283 ActionDest: 301 Annotation: Subtype: Link Rect: 287, 171, 294, 183 ActionDest: 259 Annotation: Subtype: Link Rect: 249, 133, 256, 145 ActionDest: 259 Annotation: Subtype: Link Rect: 523, 95, 529, 107 ActionDest: 259 Page: Sequence: 11 Annotations: Annotation: Subtype: Link Rect: 214, 117, 221, 127 ActionDest: 198 Page: Sequence: 12 Annotations: Annotation: Subtype: Link Rect: 195, 733, 202, 745 ActionDest: 230 Annotation: Subtype: Link Rect: 544, 471, 556, 480 ActionDest: 301 Annotation: Subtype: Link Rect: 497, 446, 509, 455 ActionDest: 298 Annotation: Subtype: Link Rect: 459, 321, 471, 330 ActionDest: 298 Annotation: Subtype: Link Rect: 330, 296, 342, 305 ActionDest: 298 Annotation: Subtype: Link Rect: 318, 283, 330, 292 ActionDest: 301 Annotation: Subtype: Link Rect: 293, 195, 305, 204 ActionDest: 301 Page: Sequence: 13 Annotations: Annotation: Subtype: Link Rect: 349, 685, 361, 694 ActionDest: 301 Annotation: Subtype: Link Rect: 364, 685, 376, 694 ActionDest: 301 Annotation: Subtype: Link Rect: 216, 660, 228, 669 ActionDest: 301 Annotation: Subtype: Link Rect: 231, 660, 243, 669 ActionDest: 301 Annotation: Subtype: Link Rect: 393, 648, 405, 657 ActionDest: 301 Annotation: Subtype: Link Rect: 204, 535, 216, 544 ActionDest: 301 Annotation: Subtype: Link Rect: 544, 522, 556, 531 ActionDest: 301 Annotation: Subtype: Link Rect: 524, 497, 536, 506 ActionDest: 301 Annotation: Subtype: Link Rect: 479, 434, 491, 443 ActionDest: 301 Annotation: Subtype: Link Rect: 473, 409, 485, 418 ActionDest: 298 Annotation: Subtype: Link Rect: 228, 346, 240, 355 ActionDest: 301 Annotation: Subtype: Link Rect: 404, 309, 416, 318 ActionDest: 301 Annotation: Subtype: Link Rect: 416, 309, 428, 318 ActionDest: 301 Annotation: Subtype: Link Rect: 193, 271, 205, 280 ActionDest: 301 Annotation: Subtype: Link Rect: 438, 158, 450, 167 ActionDest: 298 Annotation: Subtype: Link Rect: 451, 158, 463, 167 ActionDest: 298 Annotation: Subtype: Link Rect: 448, 133, 460, 142 ActionDest: 301 Annotation: Subtype: Link Rect: 222, 108, 234, 117 ActionDest: 301 Page: Sequence: 14 Annotations: Annotation: Subtype: Link Rect: 189, 647, 201, 657 ActionDest: 304 Annotation: Subtype: Link Rect: 396, 622, 408, 631 ActionDest: 304 Annotation: Subtype: Link Rect: 544, 622, 556, 631 ActionDest: 304 Annotation: Subtype: Link Rect: 204, 371, 216, 380 ActionDest: 301 Annotation: Subtype: Link Rect: 536, 269, 560, 280 Annotation: Subtype: Link Rect: 165, 257, 371, 268 Page: Sequence: 15 Annotations: Annotation: Subtype: Link Rect: 291, 386, 329, 396 Annotation: Subtype: Link Rect: 140, 363, 178, 373 Annotation: Subtype: Link Rect: 184, 363, 220, 373 Annotation: Subtype: Link Rect: 59, 340, 96, 350 Annotation: Subtype: Link Rect: 102, 340, 138, 350 Annotation: Subtype: Link Rect: 59, 317, 96, 327 Annotation: Subtype: Link Rect: 102, 317, 138, 327 Annotation: Subtype: Link Rect: 434, 293, 472, 304 Annotation: Subtype: Link Rect: 478, 293, 514, 304 Annotation: Subtype: Link Rect: 387, 270, 424, 281 Annotation: Subtype: Link Rect: 301, 247, 338, 258 Annotation: Subtype: Link Rect: 59, 214, 96, 224 Annotation: Subtype: Link Rect: 455, 190, 493, 201 Annotation: Subtype: Link Rect: 210, 155, 247, 166 Annotation: Subtype: Link Rect: 282, 121, 319, 132 Annotation: Subtype: Link Rect: 326, 121, 362, 132 Annotation: Subtype: Link Rect: 416, 98, 454, 109 Annotation: Subtype: Link Rect: 460, 98, 496, 109 Annotation: Subtype: Link Rect: 59, 64, 96, 74 Page: Sequence: 16 Annotations: Annotation: Subtype: Link Rect: 118, 699, 155, 710 Annotation: Subtype: Link Rect: 161, 699, 197, 710 Annotation: Subtype: Link Rect: 86, 665, 124, 675 Annotation: Subtype: Link Rect: 130, 665, 166, 675 Annotation: Subtype: Link Rect: 186, 630, 224, 640 Annotation: Subtype: Link Rect: 442, 606, 480, 617 Annotation: Subtype: Link Rect: 104, 561, 142, 571 Annotation: Subtype: Link Rect: 253, 526, 291, 537 Annotation: Subtype: Link Rect: 305, 503, 343, 514 Annotation: Subtype: Link Rect: 399, 480, 436, 491 Annotation: Subtype: Link Rect: 443, 480, 479, 491 Annotation: Subtype: Link Rect: 192, 458, 230, 468 Annotation: Subtype: Link Rect: 104, 435, 141, 445 Annotation: Subtype: Link Rect: 227, 411, 264, 422 Annotation: Subtype: Link Rect: 271, 411, 307, 422 Annotation: Subtype: Link Rect: 412, 388, 450, 399 Annotation: Subtype: Link Rect: 456, 388, 492, 399 Annotation: Subtype: Link Rect: 104, 354, 141, 364 Annotation: Subtype: Link Rect: 122, 320, 159, 330 Annotation: Subtype: Link Rect: 95, 285, 132, 295 Annotation: Subtype: Link Rect: 456, 261, 493, 272 Annotation: Subtype: Link Rect: 460, 238, 498, 249 Annotation: Subtype: Link Rect: 168, 216, 206, 226 Annotation: Subtype: Link Rect: 212, 216, 248, 226 Annotation: Subtype: Link Rect: 250, 181, 287, 192 Annotation: Subtype: Link Rect: 293, 181, 329, 192 Annotation: Subtype: Link Rect: 59, 147, 96, 157 Annotation: Subtype: Link Rect: 200, 113, 238, 123 Annotation: Subtype: Link Rect: 154, 90, 191, 100 Annotation: Subtype: Link Rect: 197, 90, 233, 100 Annotation: Subtype: Link Rect: 59, 67, 96, 77 Annotation: Subtype: Link Rect: 102, 67, 138, 77 Page: Sequence: 17 Annotations: Annotation: Subtype: Link Rect: 108, 711, 146, 721 Annotation: Subtype: Link Rect: 190, 687, 227, 698 Annotation: Subtype: Link Rect: 59, 653, 96, 664 Checksum: b5b74283 Type: CRC32 Checksum: 4c2936024c1533f1adf888795b01125e Type: MD5 Checksum: da8c88f1448cf98746f8c89018a47abbc399973a Type: SHA-1 Checksum: c2b96459b8724dc490779a1307bc9bca024fce4fe97ecfcabd1b14b41fca0d5d Type: SHA-256