ae12546ec2b3114809758d98a9e5007d pmc_34680482.pdf fe6bf8d44feb5e6c21419a537f089b1da4a34382 pmc_34680482.pdf 91260e2ba57441b62dfa233a673a8af1eac288946e73664a42eb823b3982df78 pmc_34680482.pdf Title: Modeling Functional Limitations, Gait Impairments, and Muscle Pathology in Alzheimer’s Disease: Studies in the 3xTg-AD Mice Subject: Gait impairments in Alzheimer’s disease (AD) result from structural and functional deficiencies that generate limitations in the performance of activities and restrictions in individual’s biopsychosocial participation. In a translational way, we have used the conceptual framework proposed by the International Classification of Disability and Health Functioning (ICF) to classify and describe the functioning and disability on gait and exploratory activity in the 3xTg-AD animal model. We developed a behavioral observation method that allows us to differentiate qualitative parameters of psychomotor performance in animals’ gait, similar to the behavioral patterns observed in humans. The functional psychomotor evaluation allows measuring various dimensions of gait and exploratory activity at different stages of disease progression in dichotomy with aging. We included male 3xTg-AD mice and their non-transgenic counterpart (NTg) of 6, 12, and 16 months of age (n = 45). Here, we present the preliminary results. The 3xTg-AD mice show more significant functional impairment in gait and exploratory activity quantitative variables. The presence of movement limitations and muscle weakness mark the functional decline related to the disease severity stages that intensify with increasing age. Motor performance in 3xTg-AD is accompanied by a series of bizarre behaviors that interfere with the trajectory, which allows us to infer poor neurological control. Additionally, signs of physical frailty accompany the functional deterioration of these animals. The use of the ICF as a conceptual framework allows the functional status to be described, facilitating its interpretation and application in the rehabilitation of people with AD. Keywords: translational neuroscience; Alzheimer’s disease; gait; muscular strength; muscular endurance; motor performance; frailty Author: Lidia Castillo-Mariqueo, M. José Pérez-García and Lydia Giménez-Llort Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Fri Oct 1 09:35:28 2021 CEST ModDate: Fri Oct 1 09:49:07 2021 CEST Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 22 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 6071939 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- AWHDWW+VnURWPalladioL Type 1 Custom yes yes no 10 0 UEGAER+URWPalladioL-Ital Type 1 Custom yes yes no 15 0 VWRVZT+URWPalladioL-Bold Type 1 Custom yes yes no 20 0 ICLLFX+URWPalladioL-Roma Type 1 Custom yes yes no 24 0 YSPQJI+VnURWPalladioL-Bold Type 1 Custom yes yes no 28 0 EBFBTV+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 36 0 MCBCCI+SourceSansRoman-Regular Type 1C Custom yes yes no 58 0 MCBCCM+SourceSansRoman.613wght Type 1C Custom yes yes no 62 0 MCBCCO+SourceSansRoman-Bold Type 1C Custom yes yes no 66 0 YIVWYZ+CMSY10 Type 1 Builtin yes yes no 78 0 QWWHCD+EURM10 Type 1 Builtin yes yes no 91 0 WTXQHS+URWPalladioL-BoldItal Type 1 Custom yes yes no 101 0 LDHCMG+VnURWPalladioL-Italic Type 1 Custom yes yes no 166 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2023-10-21 02:41:22 CEST RepresentationInformation: pmc_34680482.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2023-03-31 12:05:55 CEST Size: 6071939 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 448 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@262b2c86 Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@371a67ec Children: Item: Title: Animals Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@5ed828d Item: Title: Experimental Design Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@50d0686 Item: Title: Behavioral Assessments Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7a3d45bd Children: Item: Title: Activity Spontaneous Gait and Exploration Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1e7c7811 Item: Title: Body Function Mobility and Muscular Strength Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77ec78b9 Item: Title: Body Structure Joints and Muscles Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1a3869f4 Item: Title: Motor Performance, Geotaxis, and Hindlimb Clasping Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@a38d7a3 Item: Title: Statistics Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77f99a05 Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@63440df3 Children: Item: Title: Activity Spontaneous Gait and Exploration Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@3aeaafa6 Item: Title: Body Function Mobility and Muscular Strength Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@76a3e297 Item: Title: Body Structure Joints and Muscles Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4d3167f4 Item: Title: Motor Performance, Geotaxis, and Hindlimb Clasping Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@ed9d034 Item: Title: Survival, Kyphosis, and Frailty Phenotype Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@6121c9d6 Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@87f383f Children: Item: Title: Activity Spontaneous Gait and Exploration Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4eb7f003 Item: Title: Body Function Mobility and Muscular Strength Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@eafc191 Item: Title: Body Structure Joints and Muscles Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@612fc6eb Item: Title: Motor Performance, Geotaxis, and Hindlimb Clasping Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1060b431 Item: Title: Survival, Kyphosis, and Frailty Phenotype Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@612679d6 Item: Title: Conclusions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@11758f2a Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@e720b71 Info: Title: Modeling Functional Limitations, Gait Impairments, and Muscle Pathology in Alzheimer’s Disease: Studies in the 3xTg-AD Mice Author: Lidia Castillo-Mariqueo, M. José Pérez-García and Lydia Giménez-Llort Subject: Gait impairments in Alzheimer’s disease (AD) result from structural and functional deficiencies that generate limitations in the performance of activities and restrictions in individual’s biopsychosocial participation. In a translational way, we have used the conceptual framework proposed by the International Classification of Disability and Health Functioning (ICF) to classify and describe the functioning and disability on gait and exploratory activity in the 3xTg-AD animal model. We developed a behavioral observation method that allows us to differentiate qualitative parameters of psychomotor performance in animals’ gait, similar to the behavioral patterns observed in humans. The functional psychomotor evaluation allows measuring various dimensions of gait and exploratory activity at different stages of disease progression in dichotomy with aging. We included male 3xTg-AD mice and their non-transgenic counterpart (NTg) of 6, 12, and 16 months of age (n = 45). Here, we present the preliminary results. The 3xTg-AD mice show more significant functional impairment in gait and exploratory activity quantitative variables. The presence of movement limitations and muscle weakness mark the functional decline related to the disease severity stages that intensify with increasing age. Motor performance in 3xTg-AD is accompanied by a series of bizarre behaviors that interfere with the trajectory, which allows us to infer poor neurological control. Additionally, signs of physical frailty accompany the functional deterioration of these animals. The use of the ICF as a conceptual framework allows the functional status to be described, facilitating its interpretation and application in the rehabilitation of people with AD. Keywords: translational neuroscience; Alzheimer’s disease; gait; muscular strength; muscular endurance; motor performance; frailty Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Fri Oct 01 09:35:28 CEST 2021 ModDate: Fri Oct 01 09:49:07 CEST 2021 ID: 0xfb7e6b057930ee5961f329ab8ef257b2, 0xfb7e6b057930ee5961f329ab8ef257b2 Filters: FilterPipeline: FlateDecode Fonts: Type1: Font: BaseFont: VWRVZT+URWPalladioL-Bold FontSubset: true FirstChar: 3 LastChar: 151 FontDescriptor: FontName: VWRVZT+URWPalladioL-Bold Flags: Symbolic FontBBox: -152, -301, 1000, 935 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: WTXQHS+URWPalladioL-BoldItal FontSubset: true FirstChar: 2 LastChar: 117 FontDescriptor: FontName: WTXQHS+URWPalladioL-BoldItal Flags: Symbolic FontBBox: -170, -300, 1073, 935 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: LDHCMG+VnURWPalladioL-Italic FontSubset: true FirstChar: 161 LastChar: 226 FontDescriptor: FontName: LDHCMG+VnURWPalladioL-Italic Flags: Symbolic FontBBox: -170, -305, 1010, 941 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: ICLLFX+URWPalladioL-Roma FontSubset: true FirstChar: 2 LastChar: 252 FontDescriptor: FontName: ICLLFX+URWPalladioL-Roma Flags: Symbolic FontBBox: -166, -283, 1021, 943 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: AWHDWW+VnURWPalladioL FontSubset: true FirstChar: 161 LastChar: 243 FontDescriptor: FontName: AWHDWW+VnURWPalladioL Flags: Symbolic FontBBox: -166, -283, 1021, 943 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: QWWHCD+EURM10 FontSubset: true FirstChar: 12 LastChar: 31 FontDescriptor: FontName: QWWHCD+EURM10 Flags: Symbolic FontBBox: -32, -243, 1060, 720 FontFile: true Font: BaseFont: YSPQJI+VnURWPalladioL-Bold FontSubset: true FirstChar: 178 LastChar: 189 FontDescriptor: FontName: YSPQJI+VnURWPalladioL-Bold Flags: Symbolic FontBBox: -152, -301, 1000, 935 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: YIVWYZ+CMSY10 FontSubset: true FirstChar: 0 LastChar: 15 FontDescriptor: FontName: YIVWYZ+CMSY10 Flags: Symbolic FontBBox: -29, -960, 1116, 775 FontFile: true Font: BaseFont: UEGAER+URWPalladioL-Ital FontSubset: true FirstChar: 39 LastChar: 151 FontDescriptor: FontName: UEGAER+URWPalladioL-Ital Flags: Symbolic FontBBox: -170, -305, 1010, 941 FontFile: true EncodingDictionary: Differences: true Pages: Page: Sequence: 1 Annotations: Annotation: Subtype: Link Rect: 35, 759, 198, 795 Annotation: Subtype: Link Rect: 515, 759, 560, 795 Annotation: Subtype: Link Rect: 264, 641, 275, 654 Annotation: Subtype: Link Rect: 410, 641, 421, 654 Annotation: Subtype: Link Rect: 117, 359, 154, 367 Annotation: Subtype: Link Rect: 35, 346, 154, 355 Annotation: Subtype: Link Rect: 35, 336, 102, 343 Annotation: Subtype: Link Rect: 35, 162, 97, 184 Annotation: Subtype: Link Rect: 153, 89, 154, 98 Annotation: Subtype: Link Rect: 35, 77, 147, 86 Annotation: Subtype: Link Rect: 261, 197, 268, 206 ActionDest: 163 Annotation: Subtype: Link Rect: 268, 197, 275, 206 ActionDest: 163 Annotation: Subtype: Link Rect: 392, 185, 399, 194 ActionDest: 163 Annotation: Subtype: Link Rect: 252, 147, 259, 156 ActionDest: 163 Annotation: Subtype: Link Rect: 259, 147, 266, 156 ActionDest: 163 Annotation: Subtype: Link Rect: 232, 122, 239, 131 ActionDest: 163 Annotation: Subtype: Link Rect: 242, 122, 249, 131 ActionDest: 163 Annotation: Subtype: Link Rect: 250, 97, 257, 106 ActionDest: 163 Annotation: Subtype: Link Rect: 258, 97, 265, 106 ActionDest: 163 Annotation: Subtype: Link Rect: 265, 97, 277, 106 ActionDest: 163 Annotation: Subtype: Link Rect: 523, 72, 535, 81 ActionDest: 163 Annotation: Subtype: Link Rect: 127, 17, 294, 28 Annotation: Subtype: Link Rect: 387, 17, 558, 28 Annotation: Subtype: Link Rect: 35, 446, 87, 465 Page: Sequence: 2 Annotations: Annotation: Subtype: Link Rect: 482, 737, 494, 746 ActionDest: 163 Annotation: Subtype: Link Rect: 359, 724, 371, 733 ActionDest: 163 Annotation: Subtype: Link Rect: 362, 699, 374, 708 ActionDest: 163 Annotation: Subtype: Link Rect: 374, 699, 386, 708 ActionDest: 163 Annotation: Subtype: Link Rect: 433, 686, 440, 695 ActionDest: 163 Annotation: Subtype: Link Rect: 441, 686, 452, 695 ActionDest: 163 Annotation: Subtype: Link Rect: 407, 661, 419, 670 ActionDest: 163 Annotation: Subtype: Link Rect: 466, 636, 478, 645 ActionDest: 163 Annotation: Subtype: Link Rect: 479, 636, 491, 645 ActionDest: 163 Annotation: Subtype: Link Rect: 331, 598, 343, 608 ActionDest: 163 Annotation: Subtype: Link Rect: 346, 599, 358, 608 ActionDest: 163 Annotation: Subtype: Link Rect: 197, 561, 209, 570 ActionDest: 163 Annotation: Subtype: Link Rect: 468, 548, 480, 557 ActionDest: 163 Annotation: Subtype: Link Rect: 259, 523, 271, 532 ActionDest: 163 Annotation: Subtype: Link Rect: 482, 511, 494, 520 ActionDest: 163 Annotation: Subtype: Link Rect: 257, 485, 269, 495 ActionDest: 163 Annotation: Subtype: Link Rect: 269, 485, 281, 495 ActionDest: 163 Annotation: Subtype: Link Rect: 199, 461, 211, 469 ActionDest: 163 Annotation: Subtype: Link Rect: 387, 435, 399, 444 ActionDest: 170 Annotation: Subtype: Link Rect: 407, 410, 419, 419 ActionDest: 163 Annotation: Subtype: Link Rect: 203, 385, 215, 394 ActionDest: 170 Annotation: Subtype: Link Rect: 230, 183, 242, 192 ActionDest: 170 Annotation: Subtype: Link Rect: 544, 170, 556, 180 ActionDest: 170 Page: Sequence: 3 Annotations: Annotation: Subtype: Link Rect: 338, 710, 350, 719 ActionDest: 170 Annotation: Subtype: Link Rect: 473, 493, 480, 505 ActionDest: 82 Annotation: Subtype: Link Rect: 359, 433, 371, 442 ActionDest: 170 Annotation: Subtype: Link Rect: 357, 87, 368, 96 ActionDest: 170 Annotation: Subtype: Link Rect: 368, 87, 379, 96 ActionDest: 170 Page: Sequence: 4 Annotations: Annotation: Subtype: Link Rect: 220, 707, 232, 716 ActionDest: 170 Annotation: Subtype: Link Rect: 450, 644, 462, 653 ActionDest: 170 Annotation: Subtype: Link Rect: 292, 619, 304, 628 ActionDest: 170 Annotation: Subtype: Link Rect: 225, 430, 237, 439 ActionDest: 170 Annotation: Subtype: Link Rect: 325, 380, 337, 389 ActionDest: 170 Annotation: Subtype: Link Rect: 338, 380, 350, 389 ActionDest: 170 Annotation: Subtype: Link Rect: 544, 317, 556, 326 ActionDest: 170 Annotation: Subtype: Link Rect: 340, 229, 352, 238 ActionDest: 170 Page: Sequence: 5 Annotations: Annotation: Subtype: Link Rect: 514, 584, 526, 593 ActionDest: 170 Annotation: Subtype: Link Rect: 269, 292, 276, 304 ActionDest: 98 Annotation: Subtype: Link Rect: 296, 216, 303, 228 ActionDest: 105 Page: Sequence: 6 Annotations: Annotation: Subtype: Link Rect: 77, 169, 83, 181 ActionDest: 98 Page: Sequence: 7 Page: Sequence: 8 Annotations: Annotation: Subtype: Link Rect: 545, 594, 552, 606 ActionDest: 105 Annotation: Subtype: Link Rect: 379, 506, 386, 518 ActionDest: 105 Annotation: Subtype: Link Rect: 283, 468, 290, 480 ActionDest: 114 Annotation: Subtype: Link Rect: 196, 368, 203, 380 ActionDest: 114 Annotation: Subtype: Link Rect: 197, 318, 204, 330 ActionDest: 114 Annotation: Subtype: Link Rect: 476, 292, 483, 304 ActionDest: 120 Annotation: Subtype: Link Rect: 375, 242, 382, 254 ActionDest: 120 Annotation: Subtype: Link Rect: 525, 192, 532, 204 ActionDest: 120 Page: Sequence: 9 Page: Sequence: 10 Page: Sequence: 11 Annotations: Annotation: Subtype: Link Rect: 339, 607, 346, 619 ActionDest: 114 Annotation: Subtype: Link Rect: 486, 556, 493, 568 ActionDest: 114 Annotation: Subtype: Link Rect: 260, 481, 267, 493 ActionDest: 130 Annotation: Subtype: Link Rect: 514, 418, 521, 430 ActionDest: 130 Annotation: Subtype: Link Rect: 545, 393, 552, 405 ActionDest: 130 Annotation: Subtype: Link Rect: 374, 343, 381, 355 ActionDest: 130 Annotation: Subtype: Link Rect: 197, 267, 204, 279 ActionDest: 130 Annotation: Subtype: Link Rect: 244, 267, 251, 279 ActionDest: 136 Annotation: Subtype: Link Rect: 253, 255, 260, 267 ActionDest: 142 Page: Sequence: 12 Page: Sequence: 13 Annotations: Annotation: Subtype: Link Rect: 263, 219, 270, 231 ActionDest: 145 Annotation: Subtype: Link Rect: 497, 169, 505, 181 ActionDest: 145 Annotation: Subtype: Link Rect: 333, 144, 340, 156 ActionDest: 145 Page: Sequence: 14 Annotations: Annotation: Subtype: Link Rect: 76, 453, 82, 465 ActionDest: 142 Annotation: Subtype: Link Rect: 212, 398, 219, 410 ActionDest: 151 Page: Sequence: 15 Page: Sequence: 16 Annotations: Annotation: Subtype: Link Rect: 76, 355, 82, 367 ActionDest: 151 Annotation: Subtype: Link Rect: 529, 225, 541, 234 ActionDest: 170 Annotation: Subtype: Link Rect: 544, 225, 556, 234 ActionDest: 170 Annotation: Subtype: Link Rect: 499, 200, 511, 209 ActionDest: 170 Annotation: Subtype: Link Rect: 512, 200, 524, 209 ActionDest: 170 Annotation: Subtype: Link Rect: 409, 174, 421, 183 ActionDest: 170 Annotation: Subtype: Link Rect: 421, 174, 433, 183 ActionDest: 170 Annotation: Subtype: Link Rect: 304, 112, 316, 121 ActionDest: 170 Annotation: Subtype: Link Rect: 487, 99, 499, 108 ActionDest: 170 Annotation: Subtype: Link Rect: 500, 99, 512, 108 ActionDest: 170 Annotation: Subtype: Link Rect: 512, 99, 524, 108 ActionDest: 170 Page: Sequence: 17 Annotations: Annotation: Subtype: Link Rect: 485, 732, 492, 744 ActionDest: 82 Annotation: Subtype: Link Rect: 422, 659, 434, 668 ActionDest: 170 Annotation: Subtype: Link Rect: 435, 659, 447, 668 ActionDest: 170 Annotation: Subtype: Link Rect: 487, 559, 499, 568 ActionDest: 170 Annotation: Subtype: Link Rect: 325, 521, 336, 530 ActionDest: 170 Annotation: Subtype: Link Rect: 497, 471, 509, 480 ActionDest: 170 Annotation: Subtype: Link Rect: 315, 446, 327, 455 ActionDest: 170 Annotation: Subtype: Link Rect: 374, 421, 386, 430 ActionDest: 170 Annotation: Subtype: Link Rect: 386, 421, 398, 430 ActionDest: 170 Annotation: Subtype: Link Rect: 401, 421, 413, 430 ActionDest: 170 Annotation: Subtype: Link Rect: 334, 358, 346, 367 ActionDest: 174 Annotation: Subtype: Link Rect: 346, 358, 358, 367 ActionDest: 174 Annotation: Subtype: Link Rect: 323, 345, 335, 354 ActionDest: 174 Annotation: Subtype: Link Rect: 358, 333, 370, 342 ActionDest: 174 Annotation: Subtype: Link Rect: 437, 283, 449, 292 ActionDest: 170 Annotation: Subtype: Link Rect: 449, 283, 461, 292 ActionDest: 170 Annotation: Subtype: Link Rect: 462, 283, 474, 292 ActionDest: 174 Annotation: Subtype: Link Rect: 415, 257, 427, 267 ActionDest: 170 Annotation: Subtype: Link Rect: 461, 220, 473, 229 ActionDest: 174 Annotation: Subtype: Link Rect: 187, 207, 199, 216 ActionDest: 174 Annotation: Subtype: Link Rect: 199, 207, 211, 216 ActionDest: 174 Annotation: Subtype: Link Rect: 209, 106, 221, 116 ActionDest: 170 Annotation: Subtype: Link Rect: 529, 81, 541, 90 ActionDest: 174 Annotation: Subtype: Link Rect: 544, 81, 556, 90 ActionDest: 174 Page: Sequence: 18 Annotations: Annotation: Subtype: Link Rect: 243, 722, 255, 731 ActionDest: 170 Annotation: Subtype: Link Rect: 256, 722, 268, 731 ActionDest: 174 Annotation: Subtype: Link Rect: 436, 685, 448, 694 ActionDest: 174 Annotation: Subtype: Link Rect: 286, 672, 298, 681 ActionDest: 174 Annotation: Subtype: Link Rect: 426, 659, 438, 669 ActionDest: 170 Annotation: Subtype: Link Rect: 259, 584, 271, 593 ActionDest: 170 Annotation: Subtype: Link Rect: 271, 584, 283, 593 ActionDest: 170 Annotation: Subtype: Link Rect: 284, 584, 296, 593 ActionDest: 170 Annotation: Subtype: Link Rect: 346, 471, 358, 480 ActionDest: 174 Annotation: Subtype: Link Rect: 203, 446, 215, 455 ActionDest: 174 Annotation: Subtype: Link Rect: 378, 370, 390, 380 ActionDest: 174 Annotation: Subtype: Link Rect: 269, 345, 281, 354 ActionDest: 174 Annotation: Subtype: Link Rect: 262, 308, 274, 317 ActionDest: 174 Annotation: Subtype: Link Rect: 277, 308, 289, 317 ActionDest: 174 Annotation: Subtype: Link Rect: 503, 308, 515, 317 ActionDest: 174 Annotation: Subtype: Link Rect: 516, 308, 528, 317 ActionDest: 174 Page: Sequence: 19 Annotations: Annotation: Subtype: Link Rect: 467, 672, 478, 681 ActionDest: 174 Annotation: Subtype: Link Rect: 481, 672, 493, 681 ActionDest: 174 Annotation: Subtype: Link Rect: 481, 621, 493, 631 ActionDest: 174 Page: Sequence: 20 Annotations: Annotation: Subtype: Link Rect: 131, 650, 169, 660 Annotation: Subtype: Link Rect: 265, 627, 302, 637 Annotation: Subtype: Link Rect: 349, 603, 387, 614 Annotation: Subtype: Link Rect: 73, 569, 110, 580 Annotation: Subtype: Link Rect: 117, 569, 153, 580 Annotation: Subtype: Link Rect: 218, 546, 256, 557 Annotation: Subtype: Link Rect: 262, 546, 298, 557 Annotation: Subtype: Link Rect: 91, 512, 128, 522 Annotation: Subtype: Link Rect: 203, 489, 241, 499 Annotation: Subtype: Link Rect: 247, 489, 283, 499 Annotation: Subtype: Link Rect: 519, 465, 556, 476 Annotation: Subtype: Link Rect: 59, 443, 97, 453 Annotation: Subtype: Link Rect: 291, 431, 328, 441 Annotation: Subtype: Link Rect: 437, 407, 474, 418 Annotation: Subtype: Link Rect: 481, 407, 517, 418 Annotation: Subtype: Link Rect: 520, 384, 557, 395 Annotation: Subtype: Link Rect: 126, 362, 164, 372 Annotation: Subtype: Link Rect: 519, 350, 557, 361 Annotation: Subtype: Link Rect: 59, 339, 95, 349 Annotation: Subtype: Link Rect: 442, 327, 479, 338 Annotation: Subtype: Link Rect: 386, 304, 424, 315 Annotation: Subtype: Link Rect: 446, 269, 484, 280 Annotation: Subtype: Link Rect: 519, 258, 557, 269 Annotation: Subtype: Link Rect: 59, 247, 95, 257 Annotation: Subtype: Link Rect: 231, 223, 269, 234 Annotation: Subtype: Link Rect: 152, 201, 190, 211 Annotation: Subtype: Link Rect: 478, 177, 515, 188 Annotation: Subtype: Link Rect: 521, 177, 557, 188 Annotation: Subtype: Link Rect: 379, 154, 416, 165 Annotation: Subtype: Link Rect: 423, 154, 459, 165 Annotation: Subtype: Link Rect: 100, 132, 137, 142 Annotation: Subtype: Link Rect: 509, 108, 547, 119 Annotation: Subtype: Link Rect: 426, 85, 463, 96 Annotation: Subtype: Link Rect: 469, 85, 505, 96 Annotation: Subtype: Link Rect: 243, 63, 281, 73 Page: Sequence: 21 Annotations: Annotation: Subtype: Link Rect: 313, 710, 350, 720 Annotation: Subtype: Link Rect: 457, 686, 495, 697 Annotation: Subtype: Link Rect: 501, 686, 537, 697 Annotation: Subtype: Link Rect: 172, 664, 209, 674 Annotation: Subtype: Link Rect: 346, 640, 383, 651 Annotation: Subtype: Link Rect: 394, 617, 432, 628 Annotation: Subtype: Link Rect: 389, 606, 426, 617 Annotation: Subtype: Link Rect: 123, 584, 160, 594 Annotation: Subtype: Link Rect: 194, 560, 232, 571 Annotation: Subtype: Link Rect: 238, 560, 274, 571 Annotation: Subtype: Link Rect: 169, 526, 206, 536 Annotation: Subtype: Link Rect: 212, 526, 248, 536 Annotation: Subtype: Link Rect: 59, 480, 97, 490 Annotation: Subtype: Link Rect: 103, 480, 139, 490 Annotation: Subtype: Link Rect: 262, 457, 300, 467 Annotation: Subtype: Link Rect: 59, 423, 97, 433 Annotation: Subtype: Link Rect: 103, 423, 139, 433 Annotation: Subtype: Link Rect: 59, 400, 97, 410 Annotation: Subtype: Link Rect: 103, 400, 139, 410 Annotation: Subtype: Link Rect: 272, 376, 310, 387 Annotation: Subtype: Link Rect: 338, 354, 375, 364 Annotation: Subtype: Link Rect: 208, 331, 245, 341 Annotation: Subtype: Link Rect: 176, 307, 214, 318 Annotation: Subtype: Link Rect: 279, 249, 316, 260 Annotation: Subtype: Link Rect: 95, 215, 133, 226 Annotation: Subtype: Link Rect: 139, 215, 175, 226 Annotation: Subtype: Link Rect: 394, 192, 431, 203 Annotation: Subtype: Link Rect: 437, 192, 474, 203 Annotation: Subtype: Link Rect: 389, 169, 426, 180 Annotation: Subtype: Link Rect: 309, 134, 347, 145 Annotation: Subtype: Link Rect: 175, 112, 213, 122 Annotation: Subtype: Link Rect: 182, 77, 220, 88 Annotation: Subtype: Link Rect: 226, 77, 262, 88 Page: Sequence: 22 Annotations: Annotation: Subtype: Link Rect: 366, 721, 404, 732 Annotation: Subtype: Link Rect: 131, 687, 169, 697 Annotation: Subtype: Link Rect: 175, 687, 211, 697 Annotation: Subtype: Link Rect: 262, 652, 300, 663 Annotation: Subtype: Link Rect: 306, 652, 342, 663 Annotation: Subtype: Link Rect: 454, 629, 491, 640 Annotation: Subtype: Link Rect: 159, 594, 196, 605 Annotation: Subtype: Link Rect: 288, 548, 326, 559 Annotation: Subtype: Link Rect: 467, 525, 505, 536 Annotation: Subtype: Link Rect: 341, 502, 379, 513 Annotation: Subtype: Link Rect: 59, 480, 97, 490 Annotation: Subtype: Link Rect: 293, 456, 330, 467 Annotation: Subtype: Link Rect: 337, 456, 373, 467 Annotation: Subtype: Link Rect: 309, 422, 347, 433 Annotation: Subtype: Link Rect: 256, 399, 294, 410 Annotation: Subtype: Link Rect: 300, 399, 336, 410 Annotation: Subtype: Link Rect: 203, 377, 240, 387 Annotation: Subtype: Link Rect: 288, 353, 326, 364 Annotation: Subtype: Link Rect: 332, 353, 368, 364 Annotation: Subtype: Link Rect: 483, 330, 520, 341 Annotation: Subtype: Link Rect: 315, 307, 352, 318 Annotation: Subtype: Link Rect: 358, 307, 394, 318 Annotation: Subtype: Link Rect: 502, 284, 539, 295 Annotation: Subtype: Link Rect: 286, 261, 323, 272 Annotation: Subtype: Link Rect: 330, 261, 366, 272 Annotation: Subtype: Link Rect: 205, 227, 243, 237 Annotation: Subtype: Link Rect: 249, 227, 285, 237 Annotation: Subtype: Link Rect: 239, 203, 276, 214 Annotation: Subtype: Link Rect: 520, 192, 557, 203 Annotation: Subtype: Link Rect: 59, 181, 95, 191 Annotation: Subtype: Link Rect: 357, 157, 395, 168 Annotation: Subtype: Link Rect: 401, 157, 437, 168 Annotation: Subtype: Link Rect: 109, 123, 146, 134 Checksum: 8733ed59 Type: CRC32 Checksum: ae12546ec2b3114809758d98a9e5007d Type: MD5 Checksum: fe6bf8d44feb5e6c21419a537f089b1da4a34382 Type: SHA-1