fb9dc72ad3b6756da3b5fa3631c047e7 pmc_8942837.pdf 68e52ac40d448168768cf8d0f015468910691215 pmc_8942837.pdf dda486cf1294e033e795f8f43907fc047e887fd11347730fd58874e4c30907ba pmc_8942837.pdf Title: Protein-conjugated microbubbles for the selective targeting of S. aureus biofilms Subject: Biofilm, 4 (2022) 100074. doi:10.1016/j.bioflm.2022.100074 Keywords: Ultrasound contrast agents,Biofilm,Staphylococcus aureus,Molecular targeting,Affimer protein,Sonobactericide,Ultrasound Author: Jack A. Caudwell Creator: Elsevier Producer: Acrobat Distiller 8.1.0 (Windows) CreationDate: Tue Mar 22 06:56:34 2022 CET ModDate: Tue Mar 22 07:17:03 2022 CET Custom Metadata: yes Metadata Stream: yes Tagged: yes UserProperties: no Suspects: no Form: none JavaScript: no Pages: 11 Encrypted: no Page size: 595.276 x 793.701 pts Page rot: 0 File size: 6498385 bytes Optimized: yes PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- JPFNMX+CharisSIL CID TrueType Identity-H yes yes yes 917 0 IMGHFH+CharisSIL-Bold CID TrueType Identity-H yes yes yes 922 0 MQDDLO+Univers Type 1C Builtin yes yes yes 927 0 NOSCDU+TeX_CM_Maths_Symbols Type 1C Builtin yes yes yes 931 0 VJTFWU+CharisSIL-Italic CID TrueType Identity-H yes yes yes 935 0 ADPENH+STIX-Regular Type 1C Custom yes yes yes 940 0 KUTIND+CharisSIL CID TrueType Identity-H yes yes yes 908 0 FRKQEV+TimesNewRomanPSMT CID TrueType Identity-H yes yes yes 395 0 ROMMKX+CharisSIL-Italic CID TrueType Identity-H yes yes yes 388 0 SSHIIE+TeX_CM_Maths_Italic Type 1C Builtin yes yes yes 72 0 QJFZWB+TeX_CM_Roman Type 1C Builtin yes yes yes 172 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-05-07 04:28:22 CEST RepresentationInformation: pmc_8942837.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2022-03-28 07:45:28 CEST Size: 6498385 Format: PDF Version: 1.7 Status: Well-Formed, but not valid SignatureMatches: PDF-hul ErrorMessage: Invalid indirect destination - referenced object 'appsec1' cannot be found ID: PDF-HUL-149 ErrorMessage: edu.harvard.hul.ois.jhove.module.pdf.PdfInvalidException: Invalid indirect destination - referenced object 'appsec1' cannot be found ID: PDF-HUL-122 Offset: 6447582 MIMEtype: application/pdf Profile: Linearized PDF, Tagged PDF PDFMetadata: Objects: 1025 FreeObjects: 1 IncrementalUpdates: 1 DocumentCatalog: PageLayout: SinglePage PageMode: UseOutlines Language: en Outlines: Item: Title: Protein-conjugated microbubbles for the selective targeting of S. aureus biofilms Destination: mk:title Children: Item: Title: 1 Introduction Destination: mk:H1_1 Item: Title: 2 Materials and methods Destination: mk:H1_2 Children: Item: Title: 2.1 Microfluidic platform for biofilm studies Destination: mk:H2_1 Item: Title: 2.2 Microfluidic biofilm cultivation Destination: mk:H2_2 Item: Title: 2.3 Fluorescent staining and biofilm imaging Destination: mk:H2_3 Item: Title: 2.4 Selection of anti-clumping factor A Affimer (AClfA1) proteins Destination: mk:H2_4 Item: Title: 2.5 Production of Affimer proteins AClfA1 and AAC Destination: mk:H2_5 Item: Title: 2.6 Isothermal titration calorimetry (ITC) Destination: mk:H2_6 Item: Title: 2.7 Polystyrene bead   Affimer conjugation Destination: mk:H2_7 Item: Title: 2.8 Lipid preparation Destination: mk:H2_8 Item: Title: 2.9 Microbubble production and sample quantification Destination: mk:H2_9 Item: Title: 2.10 Microbubble   Affimer attachment and washing Destination: mk:H2_10 Item: Title: 2.11 Affimer   microbubble (and Affimer   bead) interactions with biofilms Destination: mk:H2_11 Item: Title: 2.12 Biofilm viability and analysis Destination: mk:H2_12 Item: Title: 2.13 Ultrasound exposure and microbubble destruction Destination: mk:H2_13 Item: Title: 3 Results Destination: mk:H1_3 Children: Item: Title: 3.1 Production of S. aureus biofilm under flow conditions Destination: mk:H2_14 Item: Title: 3.2 Affimer screening and selection Destination: mk:H2_15 Item: Title: 3.3 Characterization of the binding kinetics of AClfA1 to clumping factor A (ClfA) Destination: mk:H2_16 Item: Title: 3.4 AClfA1 and AAC functionalized micro-bead binding to biofilms Destination: mk:H2_17 Item: Title: 3.5 AClfA1 and AAC functionalized MB binding to biofilms Destination: mk:H2_18 Item: Title: 3.6 Effect of targeted microbubbles and ultrasound exposure on S. aureus biofilms Destination: mk:H2_19 Item: Title: 4 Discussion Destination: mk:H1_4 Children: Item: Title: 4.1 Microfluidics for biofilm study Destination: mk:H2_20 Item: Title: 4.2 AClfA1 and ClfA binding kinetics Destination: mk:H2_21 Item: Title: 4.3 AClfA1 and AAC binding when conjugated to beads and microbubbles Destination: mk:H2_22 Item: Title: 4.4 Effect of microbubble destruction on biofilm viability Destination: mk:H2_23 Item: Title: 4.5 Limitations of the study Destination: mk:H2_24 Item: Title: 5 Conclusions Destination: mk:H1_5 Item: Title: Funding Destination: mk:H1_6 Item: Title: Data availability statement Destination: mk:H1_7 Item: Title: CRediT authorship contribution statement Destination: mk:H1_8 Item: Title: Declaration of competing interest Destination: mk:HH1_2 Item: Title: Acknowledgments Destination: mk:HH1_3 Item: Title: Appendix A Supplementary data Destination: mk:H1_9 Item: Title: References Destination: mk:HH2_1 Info: Title: Protein-conjugated microbubbles for the selective targeting of S. aureus biofilms Author: Jack A. Caudwell Subject: Biofilm, 4 (2022) 100074. doi:10.1016/j.bioflm.2022.100074 Keywords: Ultrasound contrast agents,Biofilm,Staphylococcus aureus,Molecular targeting,Affimer protein,Sonobactericide,Ultrasound Creator: Elsevier Producer: Acrobat Distiller 8.1.0 (Windows) CreationDate: Tue Mar 22 06:56:34 CET 2022 ModDate: Tue Mar 22 07:17:03 CET 2022 ID: 0xa9bdc0982999f84069afb5bc14d1a29d, 0x35d734a83716aceab1290d425828e82e Filters: FilterPipeline: FlateDecode FilterPipeline: DCTDecode Images: Image: NisoImageMetadata: FormatName: image/jpg CompressionScheme: JPEG ImageWidth: 236 ImageHeight: 298 BitsPerSample: 8 BitsPerSampleUnit: integer Image: NisoImageMetadata: FormatName: image/jpg CompressionScheme: JPEG ImageWidth: 248 ImageHeight: 271 BitsPerSample: 8 BitsPerSampleUnit: integer Fonts: Type0: Font: BaseFont: JPFNMX+CharisSIL Encoding: Identity-H ToUnicode: true Font: BaseFont: VJTFWU+CharisSIL-Italic Encoding: Identity-H ToUnicode: true Font: BaseFont: IMGHFH+CharisSIL-Bold Encoding: Identity-H ToUnicode: true Font: BaseFont: FRKQEV+TimesNewRomanPSMT Encoding: Identity-H ToUnicode: true Type1: Font: BaseFont: NOSCDU+TeX_CM_Maths_Symbols FontSubset: true FirstChar: 1 LastChar: 138 FontDescriptor: FontName: NOSCDU+TeX_CM_Maths_Symbols Flags: Symbolic FontBBox: -50, -180, 1000, 720 FontFile3: true ToUnicode: true Font: BaseFont: SSHIIE+TeX_CM_Maths_Italic FontSubset: true FirstChar: 1 LastChar: 127 FontDescriptor: FontName: SSHIIE+TeX_CM_Maths_Italic Flags: Symbolic FontBBox: -50, -180, 1000, 720 FontFile3: true ToUnicode: true Font: BaseFont: ADPENH+STIX-Regular FontSubset: true FirstChar: 1 LastChar: 9 FontDescriptor: FontName: ADPENH+STIX-Regular Flags: Nonsymbolic FontBBox: -970, -486, 2208, 1023 FontFile3: true EncodingDictionary: BaseEncoding: WinAnsiEncoding Differences: true ToUnicode: true Font: BaseFont: QJFZWB+TeX_CM_Roman FontSubset: true FirstChar: 1 LastChar: 128 FontDescriptor: FontName: QJFZWB+TeX_CM_Roman Flags: Symbolic FontBBox: -50, -180, 1000, 720 FontFile3: true ToUnicode: true Font: BaseFont: MQDDLO+Univers FontSubset: true FirstChar: 32 LastChar: 251 FontDescriptor: FontName: MQDDLO+Univers Flags: Symbolic FontBBox: -50, -180, 1000, 720 FontFile3: true ToUnicode: true CIDFontType2: Font: BaseFont: JPFNMX+CharisSIL CIDSystemInfo: Registry: Adobe Registry: Identity Supplement: 0 FontDescriptor: FontName: JPFNMX+CharisSIL Flags: Nonsymbolic FontBBox: -692, -533, 1623, 1319 FontFile2: true Font: BaseFont: VJTFWU+CharisSIL-Italic CIDSystemInfo: Registry: Adobe Registry: Identity Supplement: 0 FontDescriptor: FontName: VJTFWU+CharisSIL-Italic Flags: Nonsymbolic, Italic FontBBox: -716, -533, 1627, 1318 FontFile2: true Font: BaseFont: IMGHFH+CharisSIL-Bold CIDSystemInfo: Registry: Adobe Registry: Identity Supplement: 0 FontDescriptor: FontName: IMGHFH+CharisSIL-Bold Flags: Nonsymbolic, ForceBold FontBBox: -720, -533, 1759, 1320 FontFile2: true Font: BaseFont: FRKQEV+TimesNewRomanPSMT CIDSystemInfo: Registry: Adobe Registry: Identity Supplement: 0 FontDescriptor: FontName: FRKQEV+TimesNewRomanPSMT Flags: Nonsymbolic FontBBox: -568, -307, 2000, 1007 FontFile2: true XMP: application/pdf 10.1016/j.bioflm.2022.100074 Elsevier B.V. Biofilm, 4 (2022) 100074. doi:10.1016/j.bioflm.2022.100074 Ultrasound contrast agents Biofilm Staphylococcus aureus Molecular targeting Affimer protein Sonobactericide Ultrasound Protein-conjugated microbubbles for the selective targeting of S. aureus biofilms Jack A. Caudwell Jordan M. Tinkler Ben R.G. Johnson Kenneth J. McDowall Fayez Alsulaimani Christian Tiede Darren C. Tomlinson Steven Freear W. Bruce Turnbull Stephen D. Evans Jonathan A.T. Sandoe journal Biofilm © 2022 Published by Elsevier B.V. 2590-2075 4 1 December 2022 2022-12-01 100074 100074 10.1016/j.bioflm.2022.100074 https://doi.org/10.1016/j.bioflm.2022.100074 2010-04-23 true 10.1016/j.bioflm.2022.100074 elsevier.com sciencedirect.com http://creativecommons.org/licenses/by-nc-nd/4.0/ VoR 7.0 10.1016/j.bioflm.2022.100074 noindex 2010-04-23 true 22nd March 2022 sciencedirect.com elsevier.com Elsevier 2022-03-22T06:17:03 2022-03-22T05:56:34 2022-03-22T06:17:03 True 22nd March 2022 Acrobat Distiller 8.1.0 (Windows) Ultrasound contrast agents,Biofilm,Staphylococcus aureus,Molecular targeting,Affimer protein,Sonobactericide,Ultrasound uuid:1b499bed-4ce8-4c0c-b682-0d58baae1cbe uuid:b6ed5266-03cb-4855-90c0-636283357f42 Pages: Page: Label: 1 Annotations: Annotation: Subtype: Link Rect: 121, 576, 127, 587 Flags: Print Destination: 893 Annotation: Subtype: Link Rect: 128, 576, 134, 587 Flags: Print Destination: 893 Annotation: Subtype: Link Rect: 135, 576, 141, 587 Flags: Print Destination: 893 Annotation: Subtype: Link Rect: 142, 571, 240, 582 Flags: Print Destination: 893 Annotation: Subtype: Link Rect: 240, 576, 246, 587 Flags: Print Destination: 893 Annotation: Subtype: Link Rect: 247, 571, 343, 582 Flags: Print Destination: 893 Annotation: Subtype: Link Rect: 343, 571, 457, 582 Flags: Print Destination: 893 Annotation: Subtype: Link Rect: 457, 576, 464, 587 Flags: Print Destination: 893 Annotation: Subtype: Link Rect: 464, 571, 471, 582 Flags: Print Destination: 893 Annotation: Subtype: Link Rect: 125, 563, 131, 574 Flags: Print Destination: 893 Annotation: Subtype: Link Rect: 132, 563, 222, 574 Flags: Print Destination: 893 Annotation: Subtype: Link Rect: 223, 558, 332, 569 Flags: Print Destination: 893 Annotation: Subtype: Link Rect: 332, 563, 338, 574 Flags: Print Destination: 893 Annotation: Subtype: Link Rect: 339, 563, 345, 574 Flags: Print Destination: 893 Annotation: Subtype: Link Rect: 346, 558, 421, 569 Flags: Print Destination: 893 Annotation: Subtype: Link Rect: 421, 558, 444, 569 Flags: Print Destination: 893 Annotation: Subtype: Link Rect: 110, 550, 116, 561 Flags: Print Destination: 893 Annotation: Subtype: Link Rect: 117, 545, 211, 556 Flags: Print Destination: 893 Annotation: Subtype: Link Rect: 211, 550, 218, 561 Flags: Print Destination: 893 Annotation: Subtype: Link Rect: 218, 545, 334, 556 Flags: Print Destination: 893 Annotation: Subtype: Link Rect: 334, 550, 341, 561 Flags: Print Destination: 893 Annotation: Subtype: Link Rect: 341, 550, 348, 561 Flags: Print Destination: 893 Annotation: Subtype: Link Rect: 349, 553, 353, 558 Flags: Print Destination: 893 Annotation: Subtype: Link Rect: 349, 545, 353, 556 Flags: Print Destination: 893 Annotation: Subtype: Link Rect: 424, 167, 437, 175 Flags: Print Destination: 166 Annotation: Subtype: Link Rect: 98, 114, 165, 121 Flags: Print Annotation: Subtype: Link Rect: 321, 722, 369, 730 Flags: Print Annotation: Subtype: Link Rect: 262, 667, 409, 675 Flags: Print Annotation: Subtype: Link Rect: 37, 87, 187, 94 Flags: Print Annotation: Subtype: Link Rect: 297, 749, 300, 757 Flags: Print Annotation: Subtype: Link Rect: 257, 748, 336, 760 Annotation: Subtype: Link Rect: 499, 616, 528, 645 Annotation: Subtype: Link Rect: 386, 59, 552, 70 Page: Label: 2 Annotations: Annotation: Subtype: Link Rect: 133, 607, 137, 615 Flags: Print Destination: 166 Annotation: Subtype: Link Rect: 191, 576, 195, 584 Flags: Print Destination: 166 Annotation: Subtype: Link Rect: 197, 576, 202, 584 Flags: Print Destination: 166 Annotation: Subtype: Link Rect: 59, 534, 63, 542 Flags: Print Destination: 166 Annotation: Subtype: Link Rect: 66, 534, 70, 542 Flags: Print Destination: 166 Annotation: Subtype: Link Rect: 151, 503, 168, 511 Flags: Print Destination: 166 Annotation: Subtype: Link Rect: 115, 429, 124, 437 Flags: Print Destination: 166 Annotation: Subtype: Link Rect: 127, 429, 136, 437 Flags: Print Destination: 166 Annotation: Subtype: Link Rect: 262, 388, 271, 395 Flags: Print Destination: 166 Annotation: Subtype: Link Rect: 274, 388, 283, 395 Flags: Print Destination: 166 Annotation: Subtype: Link Rect: 214, 335, 236, 343 Flags: Print Destination: 166 Annotation: Subtype: Link Rect: 98, 304, 120, 312 Flags: Print Destination: 166 Annotation: Subtype: Link Rect: 111, 272, 120, 280 Flags: Print Destination: 166 Annotation: Subtype: Link Rect: 103, 252, 125, 259 Flags: Print Destination: 166 Annotation: Subtype: Link Rect: 261, 231, 283, 239 Flags: Print Destination: 166 Annotation: Subtype: Link Rect: 174, 147, 183, 155 Flags: Print Destination: 166 Annotation: Subtype: Link Rect: 219, 126, 228, 134 Flags: Print Destination: 166 Annotation: Subtype: Link Rect: 253, 105, 275, 113 Flags: Print Destination: 166 Annotation: Subtype: Link Rect: 215, 63, 224, 71 Flags: Print Destination: 166 Annotation: Subtype: Link Rect: 390, 699, 399, 707 Flags: Print Destination: 299 Annotation: Subtype: Link Rect: 511, 699, 530, 707 Flags: Print Destination: 1 Annotation: Subtype: Link Rect: 309, 573, 331, 581 Flags: Print Destination: 299 Annotation: Subtype: Link Rect: 534, 136, 542, 143 Flags: Print Destination: 299 Page: Label: 3 Annotations: Annotation: Subtype: Link Rect: 127, 656, 148, 663 Flags: Print Destination: 32 Annotation: Subtype: Link Rect: 158, 656, 195, 663 Flags: Print Destination: null Annotation: Subtype: Link Rect: 163, 635, 172, 643 Flags: Print Destination: 299 Annotation: Subtype: Link Rect: 248, 366, 285, 374 Flags: Print Destination: null Annotation: Subtype: Link Rect: 417, 730, 426, 738 Flags: Print Destination: 299 Annotation: Subtype: Link Rect: 429, 730, 438, 738 Flags: Print Destination: 299 Annotation: Subtype: Link Rect: 333, 433, 352, 441 Flags: Print Destination: 32 Annotation: Subtype: Link Rect: 453, 216, 486, 223 Flags: Print Destination: null Page: Label: 4 Annotations: Annotation: Subtype: Link Rect: 144, 699, 153, 707 Flags: Print Destination: 299 Annotation: Subtype: Link Rect: 190, 647, 228, 655 Flags: Print Destination: null Annotation: Subtype: Link Rect: 238, 647, 247, 655 Flags: Print Destination: 299 Annotation: Subtype: Link Rect: 149, 636, 153, 644 Flags: Print Destination: 104 Annotation: Subtype: Link Rect: 173, 573, 194, 581 Flags: Print Destination: 1 Annotation: Subtype: Link Rect: 157, 563, 161, 571 Flags: Print Destination: 104 Annotation: Subtype: Link Rect: 237, 322, 246, 330 Flags: Print Destination: 299 Annotation: Subtype: Link Rect: 149, 312, 153, 320 Flags: Print Destination: 66 Annotation: Subtype: Link Rect: 309, 626, 318, 634 Flags: Print Destination: 299 Annotation: Subtype: Link Rect: 366, 563, 375, 571 Flags: Print Destination: 299 Annotation: Subtype: Link Rect: 543, 500, 552, 508 Flags: Print Destination: 299 Annotation: Subtype: Link Rect: 344, 427, 364, 435 Flags: Print Destination: 1 Annotation: Subtype: Link Rect: 306, 92, 326, 100 Flags: Print Destination: 32 Page: Label: 5 Annotations: Annotation: Subtype: Link Rect: 95, 689, 104, 696 Flags: Print Destination: 299 Annotation: Subtype: Link Rect: 72, 594, 108, 602 Flags: Print Destination: null Annotation: Subtype: Link Rect: 37, 573, 58, 581 Flags: Print Destination: 32 Annotation: Subtype: Link Rect: 480, 657, 500, 665 Flags: Print Destination: 32 Annotation: Subtype: Link Rect: 309, 500, 345, 508 Flags: Print Destination: null Annotation: Subtype: Link Rect: 512, 436, 536, 444 Flags: Print Destination: 32 Annotation: Subtype: Link Rect: 306, 426, 317, 434 Flags: Print Destination: null Annotation: Subtype: Link Rect: 546, 394, 555, 402 Flags: Print Destination: 299 Annotation: Subtype: Link Rect: 306, 384, 315, 392 Flags: Print Destination: 299 Annotation: Subtype: Link Rect: 323, 384, 342, 392 Flags: Print Destination: 32 Annotation: Subtype: Link Rect: 437, 374, 457, 381 Flags: Print Destination: 32 Annotation: Subtype: Link Rect: 334, 342, 343, 350 Flags: Print Destination: 299 Annotation: Subtype: Link Rect: 345, 342, 354, 350 Flags: Print Destination: 299 Annotation: Subtype: Link Rect: 360, 253, 399, 261 Flags: Print Destination: null Annotation: Subtype: Link Rect: 337, 123, 346, 131 Flags: Print Destination: 299 Annotation: Subtype: Link Rect: 348, 123, 357, 131 Flags: Print Destination: 299 Annotation: Subtype: Link Rect: 410, 113, 430, 121 Flags: Print Destination: 94 Annotation: Subtype: Link Rect: 404, 81, 424, 89 Flags: Print Destination: 94 Page: Label: 6 Page: Label: 7 Annotations: Annotation: Subtype: Link Rect: 58, 730, 78, 738 Flags: Print Destination: 94 Annotation: Subtype: Link Rect: 37, 643, 57, 651 Flags: Print Destination: 104 Annotation: Subtype: Link Rect: 115, 632, 137, 640 Flags: Print Destination: null Annotation: Subtype: Link Rect: 151, 545, 160, 553 Flags: Print Destination: 166 Annotation: Subtype: Link Rect: 162, 545, 171, 553 Flags: Print Destination: 166 Annotation: Subtype: Link Rect: 173, 545, 182, 553 Flags: Print Destination: 299 Annotation: Subtype: Link Rect: 185, 545, 194, 553 Flags: Print Destination: 299 Annotation: Subtype: Link Rect: 37, 409, 58, 417 Flags: Print Destination: 128 Annotation: Subtype: Link Rect: 408, 720, 473, 728 Flags: Print Destination: null Annotation: Subtype: Link Rect: 384, 593, 393, 601 Flags: Print Destination: 166 Annotation: Subtype: Link Rect: 339, 541, 348, 549 Flags: Print Destination: 166 Annotation: Subtype: Link Rect: 350, 541, 359, 549 Flags: Print Destination: 299 Annotation: Subtype: Link Rect: 362, 541, 371, 549 Flags: Print Destination: 299 Annotation: Subtype: Link Rect: 497, 499, 533, 507 Flags: Print Destination: null Annotation: Subtype: Link Rect: 485, 488, 521, 496 Flags: Print Destination: null Annotation: Subtype: Link Rect: 409, 399, 418, 407 Flags: Print Destination: 299 Annotation: Subtype: Link Rect: 420, 399, 429, 407 Flags: Print Destination: 299 Page: Label: 8 Annotations: Annotation: Subtype: Link Rect: 224, 213, 233, 221 Flags: Print Destination: 166 Annotation: Subtype: Link Rect: 235, 213, 244, 221 Flags: Print Destination: 299 Annotation: Subtype: Link Rect: 251, 81, 288, 89 Flags: Print Destination: null Annotation: Subtype: Link Rect: 543, 192, 552, 200 Flags: Print Destination: 299 Annotation: Subtype: Link Rect: 473, 181, 525, 189 Flags: Print Destination: 94 Annotation: Subtype: Link Rect: 425, 81, 434, 89 Flags: Print Destination: 299 Annotation: Subtype: Link Rect: 437, 81, 446, 89 Flags: Print Destination: 299 Page: Label: 9 Annotations: Annotation: Subtype: Link Rect: 243, 699, 252, 707 Flags: Print Destination: 166 Annotation: Subtype: Link Rect: 255, 699, 264, 707 Flags: Print Destination: 166 Annotation: Subtype: Link Rect: 266, 699, 275, 707 Flags: Print Destination: 166 Annotation: Subtype: Link Rect: 277, 699, 286, 707 Flags: Print Destination: 299 Annotation: Subtype: Link Rect: 37, 689, 46, 696 Flags: Print Destination: 299 Annotation: Subtype: Link Rect: 67, 636, 89, 644 Flags: Print Destination: 299 Annotation: Subtype: Link Rect: 157, 573, 177, 581 Flags: Print Destination: 128 Annotation: Subtype: Link Rect: 194, 573, 258, 581 Flags: Print Destination: null Annotation: Subtype: Link Rect: 65, 532, 74, 540 Flags: Print Destination: 166 Annotation: Subtype: Link Rect: 76, 532, 85, 540 Flags: Print Destination: 166 Annotation: Subtype: Link Rect: 88, 532, 97, 540 Flags: Print Destination: 299 Annotation: Subtype: Link Rect: 99, 532, 108, 540 Flags: Print Destination: 299 Annotation: Subtype: Link Rect: 334, 615, 343, 623 Flags: Print Destination: 166 Annotation: Subtype: Link Rect: 345, 615, 354, 623 Flags: Print Destination: 166 Annotation: Subtype: Link Rect: 356, 615, 365, 623 Flags: Print Destination: 299 Annotation: Subtype: Link Rect: 439, 511, 542, 519 Flags: Print Annotation: Subtype: Link Rect: 543, 113, 557, 121 Flags: Print Annotation: Subtype: Link Rect: 306, 102, 412, 110 Flags: Print Page: Label: 10 Annotations: Annotation: Subtype: Link Rect: 81, 657, 184, 665 Flags: Print Annotation: Subtype: Link Rect: 247, 615, 288, 623 Flags: Print Annotation: Subtype: Link Rect: 37, 605, 163, 613 Flags: Print Annotation: Subtype: Link Rect: 253, 526, 286, 532 Flags: Print Annotation: Subtype: Link Rect: 53, 518, 134, 525 Flags: Print Annotation: Subtype: Link Rect: 68, 486, 195, 493 Flags: Print Annotation: Subtype: Link Rect: 232, 470, 277, 477 Flags: Print Annotation: Subtype: Link Rect: 53, 462, 112, 469 Flags: Print Annotation: Subtype: Link Rect: 53, 454, 286, 461 Flags: Print Annotation: Subtype: Link Rect: 53, 446, 255, 453 Flags: Print Annotation: Subtype: Link Rect: 53, 438, 286, 445 Flags: Print Annotation: Subtype: Link Rect: 53, 430, 284, 437 Flags: Print Annotation: Subtype: Link Rect: 53, 422, 172, 429 Flags: Print Annotation: Subtype: Link Rect: 151, 407, 278, 413 Flags: Print Annotation: Subtype: Link Rect: 79, 391, 187, 397 Flags: Print Annotation: Subtype: Link Rect: 177, 375, 249, 381 Flags: Print Annotation: Subtype: Link Rect: 53, 367, 145, 373 Flags: Print Annotation: Subtype: Link Rect: 53, 343, 199, 349 Flags: Print Annotation: Subtype: Link Rect: 253, 327, 286, 333 Flags: Print Annotation: Subtype: Link Rect: 54, 319, 148, 325 Flags: Print Annotation: Subtype: Link Rect: 98, 295, 229, 301 Flags: Print Annotation: Subtype: Link Rect: 185, 263, 268, 269 Flags: Print Annotation: Subtype: Link Rect: 54, 255, 74, 261 Flags: Print Annotation: Subtype: Link Rect: 237, 223, 283, 230 Flags: Print Annotation: Subtype: Link Rect: 54, 215, 150, 222 Flags: Print Annotation: Subtype: Link Rect: 129, 191, 237, 198 Flags: Print Annotation: Subtype: Link Rect: 264, 167, 286, 174 Flags: Print Annotation: Subtype: Link Rect: 54, 159, 162, 166 Flags: Print Annotation: Subtype: Link Rect: 166, 144, 257, 150 Flags: Print Annotation: Subtype: Link Rect: 54, 136, 109, 142 Flags: Print Annotation: Subtype: Link Rect: 54, 128, 286, 134 Flags: Print Annotation: Subtype: Link Rect: 54, 120, 262, 126 Flags: Print Annotation: Subtype: Link Rect: 54, 112, 264, 118 Flags: Print Annotation: Subtype: Link Rect: 214, 96, 286, 102 Flags: Print Annotation: Subtype: Link Rect: 54, 88, 112, 94 Flags: Print Annotation: Subtype: Link Rect: 264, 72, 286, 78 Flags: Print Annotation: Subtype: Link Rect: 54, 64, 182, 70 Flags: Print Annotation: Subtype: Link Rect: 323, 732, 549, 738 Flags: Print Annotation: Subtype: Link Rect: 323, 724, 337, 730 Flags: Print Annotation: Subtype: Link Rect: 323, 716, 554, 722 Flags: Print Annotation: Subtype: Link Rect: 323, 708, 555, 714 Flags: Print Annotation: Subtype: Link Rect: 323, 700, 542, 706 Flags: Print Annotation: Subtype: Link Rect: 323, 692, 555, 698 Flags: Print Annotation: Subtype: Link Rect: 323, 684, 555, 690 Flags: Print Annotation: Subtype: Link Rect: 323, 676, 358, 682 Flags: Print Annotation: Subtype: Link Rect: 516, 652, 549, 658 Flags: Print Annotation: Subtype: Link Rect: 323, 644, 428, 651 Flags: Print Annotation: Subtype: Link Rect: 323, 636, 555, 643 Flags: Print Annotation: Subtype: Link Rect: 323, 628, 544, 635 Flags: Print Annotation: Subtype: Link Rect: 323, 620, 476, 627 Flags: Print Annotation: Subtype: Link Rect: 344, 596, 477, 603 Flags: Print Annotation: Subtype: Link Rect: 414, 572, 532, 579 Flags: Print Annotation: Subtype: Link Rect: 364, 541, 496, 547 Flags: Print Annotation: Subtype: Link Rect: 479, 517, 550, 523 Flags: Print Annotation: Subtype: Link Rect: 323, 509, 385, 515 Flags: Print Annotation: Subtype: Link Rect: 533, 485, 555, 491 Flags: Print Annotation: Subtype: Link Rect: 323, 477, 431, 483 Flags: Print Annotation: Subtype: Link Rect: 504, 453, 550, 459 Flags: Print Annotation: Subtype: Link Rect: 323, 445, 386, 451 Flags: Print Annotation: Subtype: Link Rect: 360, 413, 458, 419 Flags: Print Annotation: Subtype: Link Rect: 323, 405, 555, 411 Flags: Print Annotation: Subtype: Link Rect: 323, 397, 545, 403 Flags: Print Annotation: Subtype: Link Rect: 323, 389, 544, 396 Flags: Print Annotation: Subtype: Link Rect: 323, 357, 543, 364 Flags: Print Annotation: Subtype: Link Rect: 323, 349, 544, 356 Flags: Print Annotation: Subtype: Link Rect: 323, 341, 417, 348 Flags: Print Annotation: Subtype: Link Rect: 495, 325, 541, 332 Flags: Print Annotation: Subtype: Link Rect: 323, 317, 410, 324 Flags: Print Annotation: Subtype: Link Rect: 355, 294, 489, 300 Flags: Print Annotation: Subtype: Link Rect: 430, 270, 544, 276 Flags: Print Annotation: Subtype: Link Rect: 323, 238, 450, 244 Flags: Print Annotation: Subtype: Link Rect: 385, 214, 535, 220 Flags: Print Annotation: Subtype: Link Rect: 510, 190, 555, 196 Flags: Print Annotation: Subtype: Link Rect: 323, 182, 398, 188 Flags: Print Annotation: Subtype: Link Rect: 493, 158, 538, 164 Flags: Print Annotation: Subtype: Link Rect: 323, 150, 414, 156 Flags: Print Annotation: Subtype: Link Rect: 381, 126, 485, 133 Flags: Print Annotation: Subtype: Link Rect: 373, 102, 502, 109 Flags: Print Annotation: Subtype: Link Rect: 411, 78, 538, 85 Flags: Print Annotation: Subtype: Link Rect: 323, 54, 453, 61 Flags: Print Page: Label: 11 Annotations: Annotation: Subtype: Link Rect: 119, 716, 250, 722 Flags: Print Annotation: Subtype: Link Rect: 54, 708, 278, 714 Flags: Print Annotation: Subtype: Link Rect: 54, 700, 151, 706 Flags: Print Annotation: Subtype: Link Rect: 112, 676, 265, 682 Flags: Print Annotation: Subtype: Link Rect: 145, 644, 256, 651 Flags: Print Annotation: Subtype: Link Rect: 214, 628, 286, 635 Flags: Print Annotation: Subtype: Link Rect: 54, 620, 105, 627 Flags: Print Annotation: Subtype: Link Rect: 54, 596, 181, 603 Flags: Print Annotation: Subtype: Link Rect: 54, 565, 158, 571 Flags: Print Annotation: Subtype: Link Rect: 214, 549, 286, 555 Flags: Print Annotation: Subtype: Link Rect: 54, 541, 94, 547 Flags: Print Annotation: Subtype: Link Rect: 174, 509, 273, 515 Flags: Print Annotation: Subtype: Link Rect: 54, 501, 68, 507 Flags: Print Annotation: Subtype: Link Rect: 116, 477, 238, 483 Flags: Print Annotation: Subtype: Link Rect: 168, 453, 284, 459 Flags: Print Annotation: Subtype: Link Rect: 104, 429, 242, 435 Flags: Print Annotation: Subtype: Link Rect: 218, 405, 263, 411 Flags: Print Annotation: Subtype: Link Rect: 54, 397, 116, 403 Flags: Print Annotation: Subtype: Link Rect: 82, 365, 187, 372 Flags: Print Annotation: Subtype: Link Rect: 54, 357, 277, 364 Flags: Print Annotation: Subtype: Link Rect: 54, 349, 79, 356 Flags: Print Annotation: Subtype: Link Rect: 89, 325, 195, 332 Flags: Print Annotation: Subtype: Link Rect: 532, 724, 554, 730 Flags: Print Annotation: Subtype: Link Rect: 323, 716, 428, 722 Flags: Print Annotation: Subtype: Link Rect: 323, 708, 555, 714 Flags: Print Annotation: Subtype: Link Rect: 323, 700, 542, 706 Flags: Print Annotation: Subtype: Link Rect: 323, 692, 541, 698 Flags: Print Annotation: Subtype: Link Rect: 323, 684, 414, 690 Flags: Print Annotation: Subtype: Link Rect: 323, 676, 555, 682 Flags: Print Annotation: Subtype: Link Rect: 323, 668, 541, 674 Flags: Print Annotation: Subtype: Link Rect: 351, 644, 460, 651 Flags: Print Annotation: Subtype: Link Rect: 431, 620, 553, 627 Flags: Print Annotation: Subtype: Link Rect: 381, 588, 531, 595 Flags: Print Annotation: Subtype: Link Rect: 449, 565, 555, 571 Flags: Print Annotation: Subtype: Link Rect: 323, 557, 330, 563 Flags: Print Annotation: Subtype: Link Rect: 466, 533, 542, 539 Flags: Print Annotation: Subtype: Link Rect: 323, 525, 377, 531 Flags: Print Annotation: Subtype: Link Rect: 389, 493, 512, 499 Flags: Print Annotation: Subtype: Link Rect: 456, 477, 528, 483 Flags: Print Annotation: Subtype: Link Rect: 323, 469, 351, 475 Flags: Print Annotation: Subtype: Link Rect: 533, 453, 555, 459 Flags: Print Annotation: Subtype: Link Rect: 323, 445, 441, 451 Flags: Print Annotation: Subtype: Link Rect: 497, 421, 542, 427 Flags: Print Annotation: Subtype: Link Rect: 323, 413, 404, 419 Flags: Print Annotation: Subtype: Link Rect: 323, 405, 524, 411 Flags: Print Annotation: Subtype: Link Rect: 362, 381, 464, 388 Flags: Print Annotation: Subtype: Link Rect: 436, 357, 524, 364 Flags: Print Annotation: Subtype: Link Rect: 323, 349, 357, 356 Flags: Print Annotation: Subtype: Link Rect: 357, 325, 496, 332 Flags: Print Checksum: b7a2f13f Type: CRC32 Checksum: fb9dc72ad3b6756da3b5fa3631c047e7 Type: MD5 Checksum: 68e52ac40d448168768cf8d0f015468910691215 Type: SHA-1 Checksum: dda486cf1294e033e795f8f43907fc047e887fd11347730fd58874e4c30907ba Type: SHA-256