9a49f89ea0d2dda5555294a03ec2973d pmc_32992968.pdf ff40020596e1cf42765e67e6ad334b996e3ad8f7 pmc_32992968.pdf e9408beac7b517b55d4566f50fbdb84b4afbb83d2f583f32fa17f273959475a6 pmc_32992968.pdf Title: In Vitro Maturation with Leukemia Inhibitory Factor Prior to the Vitrification of Bovine Oocytes Improves Their Embryo Developmental Potential and Gene Expression in Oocytes and Embryos Subject: Oocyte cryopreservation has a significant impact on subsequent embryonic development. Herein, we investigated whether supplementing in vitro maturation medium with Leukemia Inhibitory Factor (LIF) prior to vitrification affects embryo development and gene expression at different embryo developmental stages. A panel of genes including maternal effect, epigenetics, apoptosis and heat stress was relatively quantified. The results show reduced cleavage rates after vitrification, regardless of the LIF treatment. Although not statistically different from control-vitrified oocytes, oocyte apoptosis and the blastocyst yield of LIF-vitrified oocytes were similar to their non-vitrified counterparts. Vitrification increased oocyte ZAR1, NPM2 and DPPA3 gene expression while its expression decreased in LIF-vitrified oocytes to similar or close levels to those of non-vitrified oocytes. With a few gene-specific exceptions, vitrification significantly increased the expression of DNMT3A, HDAC1, KAT2A, BAX and BCL2L1 in oocytes and most stages of embryo development, while comparable expression patterns for these genes were observed between LIF-vitrified and non-vitrified groups. Vitrification increased HSPA1A expression in oocytes and HSP90AA1 in 2-cell embryos. Our data suggest that vitrification triggers stage-specific changes in gene expression throughout embryonic development. However, the inclusion of LIF in the IVM medium prior to vitrification stimulates blastocyst development and several other developmental parameters and induces oocytes and embryos to demonstrate gene expression patterns similar to those derived from non-vitrified oocytes. Keywords: oocyte; cryopreservation; cow; interleukin; cytokine; maternal effect gene; epigenetics; apoptosis; heat shock protein; maternal–embryonic transition; cell count; apoptosis; inner cell mass Author: Meritxell Vendrell-Flotats, Tania García-Martínez, Iris Martínez-Rodero, Manel Lopez-Bejar, Jonathan LaMarre, Marc Yeste and Teresa Mogas Creator: LaTeX with hyperref package Producer: pdfTeX-1.40.17 CreationDate: Fri Sep 25 15:33:35 2020 CEST ModDate: Fri Sep 25 15:33:35 2020 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: 6309121 bytes Optimized: no PDF version: 1.5 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- VCQLQJ+URWPalladioL-Ital Type 1 Custom yes yes no 131 0 ONWPPP+URWPalladioL-Bold Type 1 Custom yes yes no 132 0 WLENKM+VnURWPalladioL-Bold Type 1 Custom yes yes no 133 0 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WinAnsi yes yes no 262 0 CAFGJF+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 263 0 CAFGJH+PalatinoLinotype TrueType WinAnsi yes yes no 264 0 CAFGJD+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 290 0 CAFGJF+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 291 0 CAFGJH+PalatinoLinotype TrueType WinAnsi yes yes no 292 0 CAFGJD+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 320 0 CAFGJF+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 321 0 CAFGJG+PalatinoLinotype CID TrueType Identity-H yes yes yes 322 0 CAFGJH+PalatinoLinotype TrueType WinAnsi yes yes no 323 0 CAGDAP+BookAntiqua TrueType WinAnsi yes yes no 351 0 CAFGJD+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 352 0 CAFGJF+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 353 0 CAFGJH+PalatinoLinotype TrueType WinAnsi yes yes no 354 0 MPUDFL+PazoMath Type 1 Builtin yes yes no 489 0 CAFGJD+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 511 0 CAFGJF+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 512 0 CAFGJG+PalatinoLinotype CID TrueType Identity-H yes yes yes 513 0 CAFGJH+PalatinoLinotype TrueType WinAnsi yes yes no 514 0 CAFGKI+TimesNewRoman CID TrueType Identity-H yes yes yes 515 0 VSHNAE+Rpcxr Type 1 Builtin yes yes no 624 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2022-07-19 02:27:47 CEST RepresentationInformation: pmc_32992968.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2020-11-02 03:37:52 CET Size: 6309121 Format: PDF Version: 1.5 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 0 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: ViewerPreferences: HideToolbar: false HideMenubar: false HideWindowUI: false FitWindow: true CenterWindow: false DisplayDocTitle: false NonFullScreenPageMode: UseNone Direction: L2R ViewArea: CropBox ViewClip: CropBox PrintArea: CropBox PageClip: CropBox PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: ぉのぴひはつふっぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳散瑩潮⸲ Item: Title: げづびふぬぴびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㈮ㄩ‾㸊⡜㌷㙜㌷㝜〰ぅててづっぴびぜ〴ぜ〰はてぜ〴ぜ〰がぉうぜ〴ぜ〰こふばばぬづねづのぴちぴどはのぜ〴ぜ〰つふひどのでぜ〴ぜ〰ぉざきぜ〴ぜ〰はのぜ〴ぜ〰ぅねぢひべはぜ〴ぜ〰いづぶづぬはばねづのぴ〬ぜ〴ぜ〰ごはぴちぬぜ〴ぜ〰ぃづぬぬぜ〴ぜ〰ぎふねぢづひ〬ぜ〴ぜ〰ぉぃきぜ〴ぜ〰ちのつぜ〴ぜ〰ごぅぜ〴ぜ〰ぃづぬぬぜ〴ぜ〰いどびぴひどぢふぴどはのぜ〴ぜ〰ちのつぜ〴ぜ〰ぁばはばぴはびどびぜ〴ぜ〰げちぴづぜ〴ぜ〰はてぜ〴ぜ〰あぬちびぴはっべびぴびぜ〴ぜ〰いづひどぶづつぜ〴ぜ〰てひはねぜ〴ぜ〰ざどぴひどてどづつ〯しちひねづつぜ〴ぜ〰くはっべぴづびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㈮㈩‾㸊⡜㌷㙜㌷㝜〰ぅててづっぴびぜ〴ぜ〰はてぜ〴ぜ〰がぉうぜ〴ぜ〰こふばばぬづねづのぴちぴどはのぜ〴ぜ〰つふひどのでぜ〴ぜ〰ぉざきぜ〴ぜ〰はのぜ〴ぜ〰ぴとづぜ〴ぜ〰ぐづひっづのぴちでづびぜ〴ぜ〰はてぜ〴ぜ〰ごさぎぅが〭ぐはびどぴどぶづぜ〴ぜ〰くはっべぴづびぜ〴ぜ〰ちてぴづひぜ〴ぜ〰ざどぴひどてどっちぴどはの〯しちひねどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㈮㌩‾㸊⡜㌷㙜㌷㝜〰ぅててづっぴびぜ〴ぜ〰はてぜ〴ぜ〰がぉうぜ〴ぜ〰こふばばぬづねづのぴちぴどはのぜ〴ぜ〰つふひどのでぜ〴ぜ〰ぉざきぜ〴ぜ〰はのぜ〴ぜ〰ぇづのづぜ〴ぜ〰ぅへばひづびびどはのぜ〴ぜ〰はてぜ〴ぜ〰きづぴちばとちびづぜ〴ぜ〰ぉぉぜ〴ぜ〰くはっべぴづびぜ〴ぜ〰ちのつぜ〴ぜ〰ぅちひぬべぜ〴ぜ〰ぅねぢひべはびぜ〴ぜ〰いづひどぶづつぜ〴ぜ〰てひはねぜ〴ぜ〰ざどぴひどてどづつ〯しちひねづつぜ〴ぜ〰くはっべぴづびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㈮㐩‾㸊⡜㌷㙜㌷㝜〰ぅててづっぴびぜ〴ぜ〰はてぜ〴ぜ〰がぉうぜ〴ぜ〰こふばばぬづねづのぴちぴどはのぜ〴ぜ〰つふひどのでぜ〴ぜ〰ぉざきぜ〴ぜ〰はのぜ〴ぜ〰ぇづのづぜ〴ぜ〰ぅへばひづびびどはのぜ〴ぜ〰はてぜ〴ぜ〰ぉのぜ〴ぜ〰ざどぴひは〭ぐひはつふっづつぜ〴ぜ〰あぬちびぴはっべびぴびぜ〴ぜ〰いづひどぶづつぜ〴ぜ〰てひはねぜ〴ぜ〰ざどぴひどてどづつ〯しちひねづつぜ〴ぜ〰くはっべぴづびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳散瑩潮⸳ Children: Item: Title: ぅててづっぴびぜ〴ぜ〰はてぜ〴ぜ〰がぉうぜ〴ぜ〰こふばばぬづねづのぴちぴどはのぜ〴ぜ〰つふひどのでぜ〴ぜ〰ぉざきぜ〴ぜ〰はのぜ〴ぜ〰ぅねぢひべはぜ〴ぜ〰いづぶづぬはばねづのぴ〬ぜ〴ぜ〰ごはぴちぬぜ〴ぜ〰ぃづぬぬぜ〴ぜ〰ぎふねぢづひ〬ぜ〴ぜ〰ぉぃきぜ〴ぜ〰ちのつぜ〴ぜ〰ごぅぜ〴ぜ〰ぃづぬぬぜ〴ぜ〰いどびぴひどぢふぴどはのぜ〴ぜ〰ちのつぜ〴ぜ〰ぁばはばぴはびどびぜ〴ぜ〰げちぴづぜ〴ぜ〰はてぜ〴ぜ〰あぬちびぴはっべびぴびぜ〴ぜ〰いづひどぶづつぜ〴ぜ〰てひはねぜ〴ぜ〰ざどぴひどてどづつ〯しちひねづつぜ〴ぜ〰くはっべぴづびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㈮㈩‾㸊⡜㌷㙜㌷㝜〰ぅててづっぴびぜ〴ぜ〰はてぜ〴ぜ〰がぉうぜ〴ぜ〰こふばばぬづねづのぴちぴどはのぜ〴ぜ〰つふひどのでぜ〴ぜ〰ぉざきぜ〴ぜ〰はのぜ〴ぜ〰ぴとづぜ〴ぜ〰ぐづひっづのぴちでづびぜ〴ぜ〰はてぜ〴ぜ〰ごさぎぅが〭ぐはびどぴどぶづぜ〴ぜ〰くはっべぴづびぜ〴ぜ〰ちてぴづひぜ〴ぜ〰ざどぴひどてどっちぴどはの〯しちひねどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㈮㌩‾㸊⡜㌷㙜㌷㝜〰ぅててづっぴびぜ〴ぜ〰はてぜ〴ぜ〰がぉうぜ〴ぜ〰こふばばぬづねづのぴちぴどはのぜ〴ぜ〰つふひどのでぜ〴ぜ〰ぉざきぜ〴ぜ〰はのぜ〴ぜ〰ぇづのづぜ〴ぜ〰ぅへばひづびびどはのぜ〴ぜ〰はてぜ〴ぜ〰きづぴちばとちびづぜ〴ぜ〰ぉぉぜ〴ぜ〰くはっべぴづびぜ〴ぜ〰ちのつぜ〴ぜ〰ぅちひぬべぜ〴ぜ〰ぅねぢひべはびぜ〴ぜ〰いづひどぶづつぜ〴ぜ〰てひはねぜ〴ぜ〰ざどぴひどてどづつ〯しちひねづつぜ〴ぜ〰くはっべぴづびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㈮㐩‾㸊⡜㌷㙜㌷㝜〰ぅててづっぴびぜ〴ぜ〰はてぜ〴ぜ〰がぉうぜ〴ぜ〰こふばばぬづねづのぴちぴどはのぜ〴ぜ〰つふひどのでぜ〴ぜ〰ぉざきぜ〴ぜ〰はのぜ〴ぜ〰ぇづのづぜ〴ぜ〰ぅへばひづびびどはのぜ〴ぜ〰はてぜ〴ぜ〰ぉのぜ〴ぜ〰ざどぴひは〭ぐひはつふっづつぜ〴ぜ〰あぬちびぴはっべびぴびぜ〴ぜ〰いづひどぶづつぜ〴ぜ〰てひはねぜ〴ぜ〰ざどぴひどてどづつ〯しちひねづつぜ〴ぜ〰くはっべぴづびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳散瑩潮⸳ Item: Title: ぅててづっぴびぜ〴ぜ〰はてぜ〴ぜ〰がぉうぜ〴ぜ〰こふばばぬづねづのぴちぴどはのぜ〴ぜ〰つふひどのでぜ〴ぜ〰ぉざきぜ〴ぜ〰はのぜ〴ぜ〰ぴとづぜ〴ぜ〰ぐづひっづのぴちでづびぜ〴ぜ〰はてぜ〴ぜ〰ごさぎぅが〭ぐはびどぴどぶづぜ〴ぜ〰くはっべぴづびぜ〴ぜ〰ちてぴづひぜ〴ぜ〰ざどぴひどてどっちぴどはの〯しちひねどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㈮㌩‾㸊⡜㌷㙜㌷㝜〰ぅててづっぴびぜ〴ぜ〰はてぜ〴ぜ〰がぉうぜ〴ぜ〰こふばばぬづねづのぴちぴどはのぜ〴ぜ〰つふひどのでぜ〴ぜ〰ぉざきぜ〴ぜ〰はのぜ〴ぜ〰ぇづのづぜ〴ぜ〰ぅへばひづびびどはのぜ〴ぜ〰はてぜ〴ぜ〰きづぴちばとちびづぜ〴ぜ〰ぉぉぜ〴ぜ〰くはっべぴづびぜ〴ぜ〰ちのつぜ〴ぜ〰ぅちひぬべぜ〴ぜ〰ぅねぢひべはびぜ〴ぜ〰いづひどぶづつぜ〴ぜ〰てひはねぜ〴ぜ〰ざどぴひどてどづつ〯しちひねづつぜ〴ぜ〰くはっべぴづびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㈮㐩‾㸊⡜㌷㙜㌷㝜〰ぅててづっぴびぜ〴ぜ〰はてぜ〴ぜ〰がぉうぜ〴ぜ〰こふばばぬづねづのぴちぴどはのぜ〴ぜ〰つふひどのでぜ〴ぜ〰ぉざきぜ〴ぜ〰はのぜ〴ぜ〰ぇづのづぜ〴ぜ〰ぅへばひづびびどはのぜ〴ぜ〰はてぜ〴ぜ〰ぉのぜ〴ぜ〰ざどぴひは〭ぐひはつふっづつぜ〴ぜ〰あぬちびぴはっべびぴびぜ〴ぜ〰いづひどぶづつぜ〴ぜ〰てひはねぜ〴ぜ〰ざどぴひどてどづつ〯しちひねづつぜ〴ぜ〰くはっべぴづびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳散瑩潮⸳ Item: Title: ぅててづっぴびぜ〴ぜ〰はてぜ〴ぜ〰がぉうぜ〴ぜ〰こふばばぬづねづのぴちぴどはのぜ〴ぜ〰つふひどのでぜ〴ぜ〰ぉざきぜ〴ぜ〰はのぜ〴ぜ〰ぇづのづぜ〴ぜ〰ぅへばひづびびどはのぜ〴ぜ〰はてぜ〴ぜ〰きづぴちばとちびづぜ〴ぜ〰ぉぉぜ〴ぜ〰くはっべぴづびぜ〴ぜ〰ちのつぜ〴ぜ〰ぅちひぬべぜ〴ぜ〰ぅねぢひべはびぜ〴ぜ〰いづひどぶづつぜ〴ぜ〰てひはねぜ〴ぜ〰ざどぴひどてどづつ〯しちひねづつぜ〴ぜ〰くはっべぴづびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㈮㐩‾㸊⡜㌷㙜㌷㝜〰ぅててづっぴびぜ〴ぜ〰はてぜ〴ぜ〰がぉうぜ〴ぜ〰こふばばぬづねづのぴちぴどはのぜ〴ぜ〰つふひどのでぜ〴ぜ〰ぉざきぜ〴ぜ〰はのぜ〴ぜ〰ぇづのづぜ〴ぜ〰ぅへばひづびびどはのぜ〴ぜ〰はてぜ〴ぜ〰ぉのぜ〴ぜ〰ざどぴひは〭ぐひはつふっづつぜ〴ぜ〰あぬちびぴはっべびぴびぜ〴ぜ〰いづひどぶづつぜ〴ぜ〰てひはねぜ〴ぜ〰ざどぴひどてどづつ〯しちひねづつぜ〴ぜ〰くはっべぴづびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳散瑩潮⸳ Item: Title: ぅててづっぴびぜ〴ぜ〰はてぜ〴ぜ〰がぉうぜ〴ぜ〰こふばばぬづねづのぴちぴどはのぜ〴ぜ〰つふひどのでぜ〴ぜ〰ぉざきぜ〴ぜ〰はのぜ〴ぜ〰ぇづのづぜ〴ぜ〰ぅへばひづびびどはのぜ〴ぜ〰はてぜ〴ぜ〰ぉのぜ〴ぜ〰ざどぴひは〭ぐひはつふっづつぜ〴ぜ〰あぬちびぴはっべびぴびぜ〴ぜ〰いづひどぶづつぜ〴ぜ〰てひはねぜ〴ぜ〰ざどぴひどてどづつ〯しちひねづつぜ〴ぜ〰くはっべぴづびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳散瑩潮⸳ Item: Title: いどびっふびびどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳散瑩潮⸴ Item: Title: きちぴづひどちぬびぜ〴ぜ〰ちのつぜ〴ぜ〰きづぴとはつびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮ㄩ‾㸊⡜㌷㙜㌷㝜〰ぃとづねどっちぬびぜ〴ぜ〰ちのつぜ〴ぜ〰こふばばぬどづひびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㈩‾㸊⡜㌷㙜㌷㝜〰くはっべぴづぜ〴ぜ〰ぃはぬぬづっぴどはのぜ〴ぜ〰ちのつぜ〴ぜ〰ぉのぜ〴ぜ〰ざどぴひはぜ〴ぜ〰きちぴふひちぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㌩‾㸊⡜㌷㙜㌷㝜〰くはっべぴづぜ〴ぜ〰ざどぴひどてどっちぴどはのぜ〴ぜ〰ちのつぜ〴ぜ〰しちひねどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獵扳散瑩潮⸴⸳⸱ Children: Item: Title: ぃとづねどっちぬびぜ〴ぜ〰ちのつぜ〴ぜ〰こふばばぬどづひびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㈩‾㸊⡜㌷㙜㌷㝜〰くはっべぴづぜ〴ぜ〰ぃはぬぬづっぴどはのぜ〴ぜ〰ちのつぜ〴ぜ〰ぉのぜ〴ぜ〰ざどぴひはぜ〴ぜ〰きちぴふひちぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㌩‾㸊⡜㌷㙜㌷㝜〰くはっべぴづぜ〴ぜ〰ざどぴひどてどっちぴどはのぜ〴ぜ〰ちのつぜ〴ぜ〰しちひねどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獵扳散瑩潮⸴⸳⸱ Item: Title: くはっべぴづぜ〴ぜ〰ぃはぬぬづっぴどはのぜ〴ぜ〰ちのつぜ〴ぜ〰ぉのぜ〴ぜ〰ざどぴひはぜ〴ぜ〰きちぴふひちぴどはのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㌩‾㸊⡜㌷㙜㌷㝜〰くはっべぴづぜ〴ぜ〰ざどぴひどてどっちぴどはのぜ〴ぜ〰ちのつぜ〴ぜ〰しちひねどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獵扳散瑩潮⸴⸳⸱ Item: Title: くはっべぴづぜ〴ぜ〰ざどぴひどてどっちぴどはのぜ〴ぜ〰ちのつぜ〴ぜ〰しちひねどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獵扳散瑩潮⸴⸳⸱ Children: Item: Title: ざどぴひどてどっちぴどはのぜ〴ぜ〰ぐひはぴはっはぬぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獵扳散瑩潮⸴⸳⸲ Item: Title: しちひねどのでぜ〴ぜ〰ぐひはぴはっはぬぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㐩‾㸊⡜㌷㙜㌷㝜〰ぉのぜ〴ぜ〰ざどぴひはぜ〴ぜ〰うづひぴどぬどぺちぴどはのぜ〴ぜ〰ちのつぜ〴ぜ〰ぅねぢひべはぜ〴ぜ〰ぃふぬぴふひづぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㔩‾㸊⡜㌷㙜㌷㝜〰いどててづひづのぴどちぬぜ〴ぜ〰こぴちどのどのでぜ〴ぜ〰はてぜ〴ぜ〰あぬちびぴはっべびぴびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㘩‾㸊⡜㌷㙜㌷㝜〰ごさぎぅがぜ〴ぜ〰ぁびびちべぜ〴ぜ〰どのぜ〴ぜ〰くはっべぴづびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㜩‾㸊⡜㌷㙜㌷㝜〰げぎぁぜ〴ぜ〰ぅへぴひちっぴどはの〬ぜ〴ぜ〰げづぶづひびづぜ〴ぜ〰ごひちのびっひどばぴどはのぜ〴ぜ〰ちのつぜ〴ぜ〰けふちのぴどぴちぴどぶづぜ〴ぜ〰げづちぬ〭ごどねづぜ〴ぜ〰ぐぃげぜ〴ぜ〰ぁのちぬべびどびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㠩‾㸊⡜㌷㙜㌷㝜〰ぅへばづひどねづのぴちぬぜ〴ぜ〰いづびどでのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獵扳散瑩潮⸴⸸⸱ Item: Title: ぉのぜ〴ぜ〰ざどぴひはぜ〴ぜ〰うづひぴどぬどぺちぴどはのぜ〴ぜ〰ちのつぜ〴ぜ〰ぅねぢひべはぜ〴ぜ〰ぃふぬぴふひづぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㔩‾㸊⡜㌷㙜㌷㝜〰いどててづひづのぴどちぬぜ〴ぜ〰こぴちどのどのでぜ〴ぜ〰はてぜ〴ぜ〰あぬちびぴはっべびぴびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㘩‾㸊⡜㌷㙜㌷㝜〰ごさぎぅがぜ〴ぜ〰ぁびびちべぜ〴ぜ〰どのぜ〴ぜ〰くはっべぴづびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㜩‾㸊⡜㌷㙜㌷㝜〰げぎぁぜ〴ぜ〰ぅへぴひちっぴどはの〬ぜ〴ぜ〰げづぶづひびづぜ〴ぜ〰ごひちのびっひどばぴどはのぜ〴ぜ〰ちのつぜ〴ぜ〰けふちのぴどぴちぴどぶづぜ〴ぜ〰げづちぬ〭ごどねづぜ〴ぜ〰ぐぃげぜ〴ぜ〰ぁのちぬべびどびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㠩‾㸊⡜㌷㙜㌷㝜〰ぅへばづひどねづのぴちぬぜ〴ぜ〰いづびどでのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獵扳散瑩潮⸴⸸⸱ Item: Title: いどててづひづのぴどちぬぜ〴ぜ〰こぴちどのどのでぜ〴ぜ〰はてぜ〴ぜ〰あぬちびぴはっべびぴびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㘩‾㸊⡜㌷㙜㌷㝜〰ごさぎぅがぜ〴ぜ〰ぁびびちべぜ〴ぜ〰どのぜ〴ぜ〰くはっべぴづびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㜩‾㸊⡜㌷㙜㌷㝜〰げぎぁぜ〴ぜ〰ぅへぴひちっぴどはの〬ぜ〴ぜ〰げづぶづひびづぜ〴ぜ〰ごひちのびっひどばぴどはのぜ〴ぜ〰ちのつぜ〴ぜ〰けふちのぴどぴちぴどぶづぜ〴ぜ〰げづちぬ〭ごどねづぜ〴ぜ〰ぐぃげぜ〴ぜ〰ぁのちぬべびどびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㠩‾㸊⡜㌷㙜㌷㝜〰ぅへばづひどねづのぴちぬぜ〴ぜ〰いづびどでのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獵扳散瑩潮⸴⸸⸱ Item: Title: ごさぎぅがぜ〴ぜ〰ぁびびちべぜ〴ぜ〰どのぜ〴ぜ〰くはっべぴづびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㜩‾㸊⡜㌷㙜㌷㝜〰げぎぁぜ〴ぜ〰ぅへぴひちっぴどはの〬ぜ〴ぜ〰げづぶづひびづぜ〴ぜ〰ごひちのびっひどばぴどはのぜ〴ぜ〰ちのつぜ〴ぜ〰けふちのぴどぴちぴどぶづぜ〴ぜ〰げづちぬ〭ごどねづぜ〴ぜ〰ぐぃげぜ〴ぜ〰ぁのちぬべびどびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㠩‾㸊⡜㌷㙜㌷㝜〰ぅへばづひどねづのぴちぬぜ〴ぜ〰いづびどでのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獵扳散瑩潮⸴⸸⸱ Item: Title: げぎぁぜ〴ぜ〰ぅへぴひちっぴどはの〬ぜ〴ぜ〰げづぶづひびづぜ〴ぜ〰ごひちのびっひどばぴどはのぜ〴ぜ〰ちのつぜ〴ぜ〰けふちのぴどぴちぴどぶづぜ〴ぜ〰げづちぬ〭ごどねづぜ〴ぜ〰ぐぃげぜ〴ぜ〰ぁのちぬべびどびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獥捴楯渮㐮㠩‾㸊⡜㌷㙜㌷㝜〰ぅへばづひどねづのぴちぬぜ〴ぜ〰いづびどでのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獵扳散瑩潮⸴⸸⸱ Item: Title: ぅへばづひどねづのぴちぬぜ〴ぜ〰いづびどでのぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獵扳散瑩潮⸴⸸⸱ Children: Item: Title: ぅててづっぴびぜ〴ぜ〰はてぜ〴ぜ〰がぉうぜ〴ぜ〰こふばばぬづねづのぴちぴどはのぜ〴ぜ〰つふひどのでぜ〴ぜ〰ぉざきぜ〴ぜ〰はのぜ〴ぜ〰ぅねぢひべはぜ〴ぜ〰いづぶづぬはばねづのぴ〬ぜ〴ぜ〰ごぃぎ〬ぜ〴ぜ〰ぉぃきぜ〴ぜ〰ちのつぜ〴ぜ〰ごぅぜ〴ぜ〰ぃづぬぬぜ〴ぜ〰いどびぴひどぢふぴどはのぜ〴ぜ〰ちのつぜ〴ぜ〰ぁばはばぴはびどびぜ〴ぜ〰げちぴづぜ〴ぜ〰はてぜ〴ぜ〰あぬちびぴはっべびぴびぜ〴ぜ〰いづひどぶづつぜ〴ぜ〰てひはねぜ〴ぜ〰ざどぴひどてどづつ〯しちひねづつぜ〴ぜ〰くはっべぴづびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獵扳散瑩潮⸴⸸⸲ Item: Title: ぅててづっぴびぜ〴ぜ〰はてぜ〴ぜ〰がぉうぜ〴ぜ〰こふばばぬづねづのぴちぴどはのぜ〴ぜ〰つふひどのでぜ〴ぜ〰ぉざきぜ〴ぜ〰はのぜ〴ぜ〰ぴとづぜ〴ぜ〰ぐづひっづのぴちでづびぜ〴ぜ〰はてぜ〴ぜ〰ごさぎぅが〭ぐはびどぴどぶづぜ〴ぜ〰くはっべぴづびぜ〴ぜ〰ちてぴづひぜ〴ぜ〰ざどぴひどてどっちぴどはの〯しちひねどのでぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獵扳散瑩潮⸴⸸⸳ Item: Title: ぅててづっぴびぜ〴ぜ〰はてぜ〴ぜ〰がぉうぜ〴ぜ〰こふばばぬづねづのぴちぴどはのぜ〴ぜ〰つふひどのでぜ〴ぜ〰ぉざきぜ〴ぜ〰はのぜ〴ぜ〰ぇづのづぜ〴ぜ〰ぅへばひづびびどはのぜ〴ぜ〰はてぜ〴ぜ〰きづぴちばとちびづぜ〴ぜ〰ぉぉぜ〴ぜ〰くはっべぴづびぜ〴ぜ〰ちのつぜ〴ぜ〰ぅちひぬべぜ〴ぜ〰ぅねぢひべはびぜ〴ぜ〰いづひどぶづつぜ〴ぜ〰てひはねぜ〴ぜ〰ざどぴひどてどづつ〯しちひねづつぜ〴ぜ〰くはっべぴづびぜ〴〩਼㰠⽓ 䝯呯 䐠⡳畢獵扳散瑩潮⸴⸸⸴ Item: Title: 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