af26c26f8ef20e5a9ddf546d3cbb6de8 pmc_34834198.pdf 66ee11e72533bcd09631c6dcb6fafac02b12aca5 pmc_34834198.pdf 62ea0bca8a309071dda6687a9004f7f6f6d50d7512843fb51dbb65fc8166a508 pmc_34834198.pdf Title: Polymeric Nanoparticles and Chitosan Gel Loading Ketorolac Tromethamine to Alleviate Pain Associated with Condyloma Acuminata during the Pre- and Post-Ablation Subject: This study describes the preparation and evaluation of two formulations, a hydrogel and a nanostructured system, containing ketorolac tromethamine as an anti-inflammatory agent for the local therapy against the inflammatory process derived from the surgical excision of Condyloma acuminata. Both formulations were physicochemically characterized. In vitro release profiles show that the nanoparticles release 92% 2.3 of the total ketorolac tromethamine encapsulated, while the chitosan gel releases 18.6% 0.2. The ex vivo permeation and distribution through human skin were also assayed and was observed how the main amount of ketorolac tromethamine is retained in the epidermis. In vivo studies were accomplished to evaluate the anti-inflammatory efficacy in mice which also involved the histological analysis to confirm the in vivo results. The nanoparticles present a significantly higher anti-inflammatory efficacy than chitosan gel. The tolerability of developed formulations was assessed by monitoring the biomechanical properties of the skin before and after application of both formulations. No statistical differences in trans-epidermal water loss and skin hydration with respect to the basal values were observed and the formulations exhibited higher anti-inflammatory activity compared to a reference ketotorlac tromethamine solution. Therefore, it can be concluded that both formulations can be proposed as outstanding candidates for offering a local anti-inflammatory therapeutical tool with potential clinical application. Keywords: PLGA nanoparticles; chitosan gel; Condyloma acuminata; ketorolac tromethamine; anti-inflammatory; topical delivery Author: Salima El Moussaoui, Ismael Abo-Horan, Lyda Halbaut, Cristina Alonso, Lluïsa Coderch, María Luisa Garduño-Ramírez, Beatriz Clares, José Luis Soriano, Ana Cristina Calpena, Francisco Fernández-Campos and Mireia Mallandrich Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Thu Nov 25 09:39:15 2021 CET ModDate: Thu Nov 25 09:42:21 2021 CET Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 19 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 3157275 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- IOJCNK+VnURWPalladioL-Bold Type 1 Custom yes yes no 10 0 DUFXDP+VnURWPalladioL Type 1 Custom yes yes no 15 0 MSSWYF+URWPalladioL-Ital Type 1 Custom yes yes no 19 0 RZYPNY+URWPalladioL-Bold Type 1 Custom yes yes no 24 0 AGIEKP+URWPalladioL-Roma Type 1 Custom yes yes no 28 0 OJYQXC+CMSY10 Type 1 Builtin yes yes no 32 0 IAFAOG+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 41 0 MCBCCI+SourceSansRoman-Regular Type 1C Custom yes yes no 69 0 MCBCCM+SourceSansRoman.613wght Type 1C Custom yes yes no 73 0 MCBCCO+SourceSansRoman-Bold Type 1C Custom yes yes no 77 0 CZKGAS+CMR10 Type 1 Builtin yes yes no 92 0 OVNEXW+EURM10 Type 1 Builtin yes yes no 96 0 ZPGVCH+CMEX10 Type 1 Builtin yes yes no 109 0 OEFIKG+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 119 0 OEFIKI+PalatinoLinotype TrueType WinAnsi yes yes no 122 0 OEFIKK+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 125 0 OEFIKG+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 135 0 OEFIKH+PalatinoLinotype CID TrueType Identity-H yes yes yes 138 0 OEFIKI+PalatinoLinotype TrueType WinAnsi yes yes no 144 0 OEFIKK+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 147 0 OEFIKG+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 155 0 OEFIKH+PalatinoLinotype CID TrueType Identity-H yes yes yes 158 0 OEFIKI+PalatinoLinotype TrueType WinAnsi yes yes no 164 0 OEFIKK+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 167 0 WBVKDJ+URWPalladioL-BoldItal Type 1 Custom yes yes no 175 0 OEFNKP+PalatinoLinotype,BoldItalic TrueType WinAnsi yes yes no 184 0 OEFIKG+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 187 0 OEFIKH+PalatinoLinotype CID TrueType Identity-H yes yes yes 190 0 OEFIKI+PalatinoLinotype TrueType WinAnsi yes yes no 196 0 OEFIKK+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 199 0 QHMKWQ+PazoMath Type 1 Builtin yes yes no 206 0 OEFNKP+PalatinoLinotype,BoldItalic TrueType WinAnsi yes yes no 213 0 OEFIKG+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 216 0 OEFIKH+PalatinoLinotype CID TrueType Identity-H yes yes yes 219 0 OEFIKI+PalatinoLinotype TrueType WinAnsi yes yes no 225 0 OEFIKK+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 228 0 XIRVNA+EURB10 Type 1 Builtin yes yes no 235 0 OEFNKP+PalatinoLinotype,BoldItalic TrueType WinAnsi yes yes no 242 0 OEFIKG+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 245 0 OEFIKI+PalatinoLinotype TrueType WinAnsi yes yes no 248 0 OEFIKK+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 251 0 OEFIKG+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 262 0 OEFIKI+PalatinoLinotype TrueType WinAnsi yes yes no 265 0 OEFIKK+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 268 0 OEFIKG+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 278 0 OEFIKI+PalatinoLinotype TrueType WinAnsi yes yes no 281 0 OEFIKK+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 284 0 PGFDEU+VnURWPalladioL-Italic Type 1 Custom yes yes no 303 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2022-03-22 02:32:46 CET RepresentationInformation: pmc_34834198.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2022-01-10 08:10:56 CET Size: 3157275 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 515 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7b69c6ba Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@46daef40 Children: Item: Title: Materials Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@12f41634 Item: Title: Preparation of Formulations Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@13c27452 Children: Item: Title: Preparation of CTS Gel Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@262b2c86 Item: Title: Preparation of NPs Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@371a67ec Item: Title: Physical Characterization of Formulations Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@5ed828d Children: Item: Title: Characterization of CTS Gel Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@50d0686 Item: Title: Characterization of the NPs Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7a3d45bd Item: Title: Stability Studies Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1e7c7811 Item: Title: In Vitro Drug Release Study Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77ec78b9 Item: Title: Ex Vivo Permeation Studies Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1a3869f4 Item: Title: HPLC Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@a38d7a3 Item: Title: In Vivo Anti-Inflammatory Efficacy Evaluation Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77f99a05 Children: Item: Title: TPA-Induced Ear Oedema in Mice Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@63440df3 Item: Title: Histological Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@3aeaafa6 Item: Title: In Vivo Tolerance Study Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@76a3e297 Item: Title: Statistical Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4d3167f4 Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@ed9d034 Children: Item: Title: Characterization of Formulations Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@6121c9d6 Children: Item: Title: Characterization of CTS Gel Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@87f383f Item: Title: Characterization of NPs Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4eb7f003 Item: Title: Stability Studies Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@eafc191 Item: Title: In Vitro Drug Release Study Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@612fc6eb Item: Title: Ex Vivo Permeation of KT through Human Skin Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1060b431 Item: Title: In Vivo Anti-Inflammatory Efficacy Evaluation Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@612679d6 Children: Item: Title: TPA-Induced Ear Oedema in Mice Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@11758f2a Item: Title: Histological Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@e720b71 Item: Title: In Vivo Tolerance Study Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1b26f7b2 Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@491cc5c9 Item: Title: Conclusions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@74ad1f1f Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@6a1aab78 Info: Title: Polymeric Nanoparticles and Chitosan Gel Loading Ketorolac Tromethamine to Alleviate Pain Associated with Condyloma Acuminata during the Pre- and Post-Ablation Author: Salima El Moussaoui, Ismael Abo-Horan, Lyda Halbaut, Cristina Alonso, Lluïsa Coderch, María Luisa Garduño-Ramírez, Beatriz Clares, José Luis Soriano, Ana Cristina Calpena, Francisco Fernández-Campos and Mireia Mallandrich Subject: This study describes the preparation and evaluation of two formulations, a hydrogel and a nanostructured system, containing ketorolac tromethamine as an anti-inflammatory agent for the local therapy against the inflammatory process derived from the surgical excision of Condyloma acuminata. Both formulations were physicochemically characterized. In vitro release profiles show that the nanoparticles release 92% 2.3 of the total ketorolac tromethamine encapsulated, while the chitosan gel releases 18.6% 0.2. The ex vivo permeation and distribution through human skin were also assayed and was observed how the main amount of ketorolac tromethamine is retained in the epidermis. In vivo studies were accomplished to evaluate the anti-inflammatory efficacy in mice which also involved the histological analysis to confirm the in vivo results. The nanoparticles present a significantly higher anti-inflammatory efficacy than chitosan gel. The tolerability of developed formulations was assessed by monitoring the biomechanical properties of the skin before and after application of both formulations. No statistical differences in trans-epidermal water loss and skin hydration with respect to the basal values were observed and the formulations exhibited higher anti-inflammatory activity compared to a reference ketotorlac tromethamine solution. Therefore, it can be concluded that both formulations can be proposed as outstanding candidates for offering a local anti-inflammatory therapeutical tool with potential clinical application. Keywords: PLGA nanoparticles; chitosan gel; Condyloma acuminata; ketorolac tromethamine; anti-inflammatory; topical delivery Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Thu Nov 25 09:39:15 CET 2021 ModDate: Thu Nov 25 09:42:21 CET 2021 ID: 0xca67772d3154c38d26679d7ef0000646, 0xca67772d3154c38d26679d7ef0000646 Filters: FilterPipeline: FlateDecode Fonts: Type1: Font: BaseFont: OJYQXC+CMSY10 FontSubset: true FirstChar: 0 LastChar: 14 FontDescriptor: FontName: OJYQXC+CMSY10 Flags: Symbolic FontBBox: -29, -960, 1116, 775 FontFile: 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: IOJCNK+VnURWPalladioL-Bold FontSubset: true FirstChar: 161 LastChar: 189 FontDescriptor: FontName: IOJCNK+VnURWPalladioL-Bold Flags: Symbolic FontBBox: -152, -301, 1000, 935 FontFile: true EncodingDictionary: Differences: true 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: ZPGVCH+CMEX10 FontSubset: true FirstChar: 18 LastChar: 19 FontDescriptor: FontName: ZPGVCH+CMEX10 Flags: Symbolic FontBBox: -24, -2960, 1454, 772 FontFile: true Font: BaseFont: QHMKWQ+PazoMath FontSubset: true FirstChar: 165 LastChar: 165 FontDescriptor: FontName: QHMKWQ+PazoMath Flags: Symbolic FontBBox: -40, -283, 878, 946 FontFile: true Font: BaseFont: DUFXDP+VnURWPalladioL FontSubset: true FirstChar: 161 LastChar: 226 FontDescriptor: FontName: DUFXDP+VnURWPalladioL Flags: Symbolic FontBBox: -166, -283, 1021, 943 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: PGFDEU+VnURWPalladioL-Italic FontSubset: true FirstChar: 129 LastChar: 226 FontDescriptor: FontName: PGFDEU+VnURWPalladioL-Italic Flags: Symbolic FontBBox: -170, -305, 1010, 941 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: MSSWYF+URWPalladioL-Ital FontSubset: true FirstChar: 2 LastChar: 148 FontDescriptor: FontName: MSSWYF+URWPalladioL-Ital Flags: Symbolic FontBBox: -170, -305, 1010, 941 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: RZYPNY+URWPalladioL-Bold FontSubset: true FirstChar: 3 LastChar: 241 FontDescriptor: FontName: RZYPNY+URWPalladioL-Bold Flags: Symbolic FontBBox: -152, -301, 1000, 935 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: AGIEKP+URWPalladioL-Roma FontSubset: true FirstChar: 2 LastChar: 252 FontDescriptor: FontName: AGIEKP+URWPalladioL-Roma Flags: Symbolic FontBBox: -166, -283, 1021, 943 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: CZKGAS+CMR10 FontSubset: true FirstChar: 40 LastChar: 61 FontDescriptor: FontName: CZKGAS+CMR10 Flags: Symbolic FontBBox: -40, -250, 1009, 750 FontFile: true Pages: Page: Sequence: 1 Annotations: Annotation: Subtype: Link Rect: 35, 759, 168, 795 Annotation: Subtype: Link Rect: 515, 759, 560, 795 Annotation: Subtype: Link Rect: 404, 641, 415, 654 Annotation: Subtype: Link Rect: 181, 628, 192, 642 Annotation: Subtype: Link Rect: 278, 628, 289, 642 Annotation: Subtype: Link Rect: 378, 628, 389, 642 Annotation: Subtype: Link Rect: 504, 628, 515, 642 Annotation: Subtype: Link Rect: 181, 616, 192, 629 Annotation: Subtype: Link Rect: 322, 616, 333, 629 Annotation: Subtype: Link Rect: 54, 346, 154, 355 Annotation: Subtype: Link Rect: 35, 334, 109, 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: 207, 105, 214, 114 ActionDest: 297 Annotation: Subtype: Link Rect: 548, 105, 555, 114 ActionDest: 297 Annotation: Subtype: Link Rect: 424, 92, 431, 102 ActionDest: 297 Annotation: Subtype: Link Rect: 519, 80, 526, 89 ActionDest: 297 Annotation: Subtype: Link Rect: 136, 17, 312, 28 Annotation: Subtype: Link Rect: 383, 17, 558, 28 Annotation: Subtype: Link Rect: 35, 494, 87, 513 Page: Sequence: 2 Annotations: Annotation: Subtype: Link Rect: 314, 712, 321, 721 ActionDest: 297 Annotation: Subtype: Link Rect: 232, 699, 239, 708 ActionDest: 297 Annotation: Subtype: Link Rect: 290, 699, 297, 708 ActionDest: 297 Annotation: Subtype: Link Rect: 490, 699, 497, 708 ActionDest: 300 Annotation: Subtype: Link Rect: 365, 674, 372, 683 ActionDest: 300 Annotation: Subtype: Link Rect: 201, 649, 208, 658 ActionDest: 297 Annotation: Subtype: Link Rect: 222, 636, 229, 645 ActionDest: 297 Annotation: Subtype: Link Rect: 307, 624, 314, 633 ActionDest: 300 Annotation: Subtype: Link Rect: 368, 586, 375, 595 ActionDest: 297 Annotation: Subtype: Link Rect: 423, 561, 435, 570 ActionDest: 300 Annotation: Subtype: Link Rect: 325, 523, 337, 532 ActionDest: 300 Annotation: Subtype: Link Rect: 445, 498, 457, 507 ActionDest: 300 Annotation: Subtype: Link Rect: 312, 473, 323, 482 ActionDest: 300 Annotation: Subtype: Link Rect: 416, 448, 428, 457 ActionDest: 300 Annotation: Subtype: Link Rect: 195, 423, 207, 432 ActionDest: 300 Annotation: Subtype: Link Rect: 208, 423, 220, 432 ActionDest: 300 Annotation: Subtype: Link Rect: 498, 398, 510, 407 ActionDest: 300 Annotation: Subtype: Link Rect: 263, 347, 275, 356 ActionDest: 300 Annotation: Subtype: Link Rect: 463, 322, 474, 331 ActionDest: 300 Annotation: Subtype: Link Rect: 531, 297, 543, 306 ActionDest: 300 Annotation: Subtype: Link Rect: 544, 297, 556, 306 ActionDest: 300 Page: Sequence: 3 Annotations: Annotation: Subtype: Link Rect: 341, 520, 353, 529 ActionDest: 300 Annotation: Subtype: Link Rect: 354, 520, 365, 529 ActionDest: 300 Page: Sequence: 4 Annotations: Annotation: Subtype: Link Rect: 207, 428, 214, 437 ActionDest: 297 Page: Sequence: 5 Annotations: Annotation: Subtype: Link Rect: 279, 367, 291, 376 ActionDest: 300 Annotation: Subtype: Link Rect: 507, 304, 514, 313 ActionDest: 300 Page: Sequence: 6 Annotations: Annotation: Subtype: Link Rect: 544, 643, 556, 652 ActionDest: 300 Page: Sequence: 7 Annotations: Annotation: Subtype: Link Rect: 219, 665, 226, 677 ActionDest: 113 Annotation: Subtype: Link Rect: 425, 218, 432, 230 ActionDest: 113 Page: Sequence: 8 Annotations: Annotation: Subtype: Link Rect: 523, 707, 530, 719 ActionDest: 129 Annotation: Subtype: Link Rect: 501, 380, 508, 394 ActionDest: 129 Page: Sequence: 9 Annotations: Annotation: Subtype: Link Rect: 287, 679, 294, 691 ActionDest: 172 Annotation: Subtype: Link Rect: 366, 280, 373, 292 ActionDest: 172 Annotation: Subtype: Link Rect: 200, 68, 207, 79 ActionDest: 203 Page: Sequence: 10 Annotations: Annotation: Subtype: Link Rect: 306, 450, 313, 463 ActionDest: 203 Annotation: Subtype: Link Rect: 350, 237, 357, 249 ActionDest: 232 Page: Sequence: 11 Annotations: Annotation: Subtype: Link Rect: 399, 490, 406, 502 ActionDest: 232 Page: Sequence: 12 Annotations: Annotation: Subtype: Link Rect: 244, 704, 251, 716 ActionDest: 256 Annotation: Subtype: Link Rect: 482, 691, 489, 703 ActionDest: 256 Annotation: Subtype: Link Rect: 199, 679, 206, 691 ActionDest: 256 Annotation: Subtype: Link Rect: 526, 679, 533, 691 ActionDest: 256 Annotation: Subtype: Link Rect: 543, 666, 550, 678 ActionDest: 256 Annotation: Subtype: Link Rect: 497, 654, 504, 666 ActionDest: 256 Annotation: Subtype: Link Rect: 253, 629, 260, 641 ActionDest: 256 Annotation: Subtype: Link Rect: 220, 249, 227, 261 ActionDest: 272 Page: Sequence: 13 Annotations: Annotation: Subtype: Link Rect: 232, 351, 239, 360 ActionDest: 297 Annotation: Subtype: Link Rect: 227, 263, 239, 272 ActionDest: 300 Annotation: Subtype: Link Rect: 242, 263, 254, 272 ActionDest: 300 Annotation: Subtype: Link Rect: 405, 125, 417, 134 ActionDest: 300 Annotation: Subtype: Link Rect: 417, 125, 429, 134 ActionDest: 300 Annotation: Subtype: Link Rect: 297, 100, 309, 109 ActionDest: 300 Annotation: Subtype: Link Rect: 302, 87, 314, 96 ActionDest: 300 Annotation: Subtype: Link Rect: 317, 87, 329, 96 ActionDest: 300 Page: Sequence: 14 Annotations: Annotation: Subtype: Link Rect: 212, 672, 224, 681 ActionDest: 300 Annotation: Subtype: Link Rect: 209, 647, 221, 656 ActionDest: 300 Annotation: Subtype: Link Rect: 260, 634, 272, 643 ActionDest: 300 Annotation: Subtype: Link Rect: 483, 584, 495, 593 ActionDest: 300 Annotation: Subtype: Link Rect: 233, 572, 245, 581 ActionDest: 300 Annotation: Subtype: Link Rect: 544, 559, 556, 568 ActionDest: 307 Annotation: Subtype: Link Rect: 201, 433, 213, 443 ActionDest: 300 Annotation: Subtype: Link Rect: 520, 433, 532, 443 ActionDest: 307 Annotation: Subtype: Link Rect: 251, 295, 263, 304 ActionDest: 307 Annotation: Subtype: Link Rect: 223, 220, 235, 229 ActionDest: 307 Annotation: Subtype: Link Rect: 236, 220, 248, 229 ActionDest: 307 Annotation: Subtype: Link Rect: 514, 183, 526, 192 ActionDest: 307 Annotation: Subtype: Link Rect: 478, 120, 490, 129 ActionDest: 307 Annotation: Subtype: Link Rect: 259, 82, 271, 91 ActionDest: 307 Page: Sequence: 15 Annotations: Annotation: Subtype: Link Rect: 245, 710, 257, 719 ActionDest: 300 Annotation: Subtype: Link Rect: 339, 710, 351, 719 ActionDest: 307 Annotation: Subtype: Link Rect: 491, 634, 503, 643 ActionDest: 300 Annotation: Subtype: Link Rect: 443, 534, 455, 543 ActionDest: 307 Annotation: Subtype: Link Rect: 480, 471, 492, 480 ActionDest: 307 Annotation: Subtype: Link Rect: 209, 433, 221, 443 ActionDest: 307 Annotation: Subtype: Link Rect: 221, 433, 233, 443 ActionDest: 307 Annotation: Subtype: Link Rect: 266, 346, 278, 355 ActionDest: 300 Annotation: Subtype: Link Rect: 278, 346, 290, 355 ActionDest: 307 Annotation: Subtype: Link Rect: 248, 120, 255, 129 ActionDest: 297 Annotation: Subtype: Link Rect: 256, 120, 268, 129 ActionDest: 307 Annotation: Subtype: Link Rect: 516, 82, 523, 91 ActionDest: 297 Page: Sequence: 16 Annotations: Annotation: Subtype: Link Rect: 524, 609, 536, 618 ActionDest: 307 Annotation: Subtype: Link Rect: 344, 546, 356, 556 ActionDest: 307 Annotation: Subtype: Link Rect: 432, 534, 444, 543 ActionDest: 307 Annotation: Subtype: Link Rect: 333, 484, 339, 493 ActionDest: 297 Annotation: Subtype: Link Rect: 340, 484, 352, 493 ActionDest: 307 Annotation: Subtype: Link Rect: 500, 320, 512, 330 ActionDest: 307 Annotation: Subtype: Link Rect: 515, 320, 527, 330 ActionDest: 307 Page: Sequence: 17 Annotations: Annotation: Subtype: Link Rect: 419, 621, 560, 632 Annotation: Subtype: Link Rect: 166, 609, 301, 620 Annotation: Subtype: Link Rect: 205, 201, 403, 212 Annotation: Subtype: Link Rect: 122, 179, 160, 189 Annotation: Subtype: Link Rect: 286, 156, 323, 166 Annotation: Subtype: Link Rect: 167, 121, 204, 131 Annotation: Subtype: Link Rect: 281, 97, 319, 108 Annotation: Subtype: Link Rect: 513, 74, 551, 85 Page: Sequence: 18 Annotations: Annotation: Subtype: Link Rect: 95, 722, 133, 732 Annotation: Subtype: Link Rect: 126, 699, 164, 709 Annotation: Subtype: Link Rect: 170, 699, 206, 709 Annotation: Subtype: Link Rect: 500, 675, 537, 686 Annotation: Subtype: Link Rect: 520, 663, 557, 674 Annotation: Subtype: Link Rect: 59, 653, 95, 663 Annotation: Subtype: Link Rect: 520, 629, 557, 640 Annotation: Subtype: Link Rect: 136, 607, 173, 617 Annotation: Subtype: Link Rect: 179, 607, 216, 617 Annotation: Subtype: Link Rect: 456, 594, 494, 605 Annotation: Subtype: Link Rect: 59, 572, 97, 582 Annotation: Subtype: Link Rect: 283, 548, 320, 559 Annotation: Subtype: Link Rect: 327, 548, 363, 559 Annotation: Subtype: Link Rect: 463, 514, 500, 525 Annotation: Subtype: Link Rect: 507, 514, 543, 525 Annotation: Subtype: Link Rect: 439, 491, 476, 502 Annotation: Subtype: Link Rect: 483, 491, 519, 502 Annotation: Subtype: Link Rect: 448, 479, 486, 490 Annotation: Subtype: Link Rect: 520, 456, 557, 467 Annotation: Subtype: Link Rect: 325, 433, 363, 444 Annotation: Subtype: Link Rect: 306, 399, 343, 410 Annotation: Subtype: Link Rect: 344, 364, 382, 375 Annotation: Subtype: Link Rect: 388, 364, 424, 375 Annotation: Subtype: Link Rect: 502, 318, 539, 329 Annotation: Subtype: Link Rect: 514, 295, 551, 306 Annotation: Subtype: Link Rect: 100, 273, 137, 283 Annotation: Subtype: Link Rect: 449, 249, 486, 260 Annotation: Subtype: Link Rect: 162, 226, 199, 237 Annotation: Subtype: Link Rect: 316, 203, 354, 214 Annotation: Subtype: Link Rect: 429, 180, 467, 191 Annotation: Subtype: Link Rect: 329, 157, 366, 168 Annotation: Subtype: Link Rect: 100, 135, 137, 145 Annotation: Subtype: Link Rect: 225, 66, 262, 76 Page: Sequence: 19 Annotations: Annotation: Subtype: Link Rect: 104, 722, 141, 732 Annotation: Subtype: Link Rect: 473, 698, 510, 709 Annotation: Subtype: Link Rect: 516, 698, 552, 709 Annotation: Subtype: Link Rect: 95, 676, 133, 686 Annotation: Subtype: Link Rect: 219, 641, 256, 651 Annotation: Subtype: Link Rect: 225, 617, 262, 628 Annotation: Subtype: Link Rect: 191, 583, 229, 594 Annotation: Subtype: Link Rect: 122, 561, 160, 571 Annotation: Subtype: Link Rect: 166, 561, 202, 571 Annotation: Subtype: Link Rect: 95, 538, 133, 548 Annotation: Subtype: Link Rect: 519, 525, 557, 536 Annotation: Subtype: Link Rect: 59, 515, 95, 525 Annotation: Subtype: Link Rect: 440, 491, 477, 502 Annotation: Subtype: Link Rect: 272, 445, 309, 456 Annotation: Subtype: Link Rect: 315, 445, 351, 456 Annotation: Subtype: Link Rect: 131, 423, 169, 433 Annotation: Subtype: Link Rect: 175, 423, 211, 433 Annotation: Subtype: Link Rect: 190, 353, 228, 364 Annotation: Subtype: Link Rect: 225, 331, 263, 341 Checksum: 4066bb30 Type: CRC32 Checksum: af26c26f8ef20e5a9ddf546d3cbb6de8 Type: MD5 Checksum: 66ee11e72533bcd09631c6dcb6fafac02b12aca5 Type: SHA-1