detection-modified-forms-cytosine-using-sensitive

Detection of Modified Forms of Cytosine Using Sensitive Immunohistochemistry

Methylation of cytosine bases (5-methylcytosine, 5mC) occurring in vertebrate genomes is usually associated with transcriptional silencing. ...

immunostaining-for-dna-modifications-computational-analysis-confocal

Immunostaining for DNA Modifications: Computational Analysis of Confocal Images

For several decades, 5-methylcytosine (5mC) has been thought to be the only DNA modification with a functional significance in metazoans. The discovery of ...

Université Libre de Bruxelles

University of Nottingham

5-&#x7532;&#x57FA;&#x80DE;&#x5627;&#x5576; (5-mC) &#x662F;&#x8868;&#x89C2;&#x9057;&#x4F20;&#x4FEE;&#x9970;&#x4E0E;&#x57FA;&#x56E0;&#x6291;&#x5236;&#x6709;&#x5173;&#x3002;&#x6700;&#x8FD1;&#x7684;&#x7814;&#x7A76;&#x8868;&#x660E;&#xFF0C;5 mC &#x53EF;&#x4EE5;&#x662F;&#x9176;&#x6C27;&#x5316;&#x6210; 5 hydroxymethylcytosine &#x548C;&#x8FDB;&#x4E00;&#x6B65;&#x7EB3;&#x5165; 5-formylcytosine &#xFF08;5-fC) &#x548C; 5-carboxylcytsine &#xFF08;5-caC)&#x3002;5 caC &#x5DF2;&#x88AB;&#x53D1;&#x73B0;&#x5728;&#x80DA;&#x80CE;&#x5E72;&#x7EC6;&#x80DE;&#x5728;&#x5C0F;&#x9F20;&#x65E9;&#x671F;&#x80DA;&#x80CE;&#x4E2D;&#xFF0C;&#x4F46;&#x6B64;&#x4FEE;&#x6539;&#x6CA1;&#x6709;&#x63A2;&#x6D4B;&#x6C34;&#x5E73;&#x8FC4;&#x4ECA;&#x62A5;&#x544A;&#x4F9B;&#x4F53;&#x7EC6;&#x80DE;&#x7EC4;&#x7EC7;&#x3002;&#x800C;&#x8BA4;&#x4E3A;&#x8BE5; 5 caC &#x53EF;&#x4EE5;&#x4F5C;&#x4E3A;&#x4E2D;&#x95F4;&#x79EF;&#x6781;&#x53BB;&#x7532;&#x57FA;&#x7684;&#x8FC7;&#x7A0B;&#x4E2D;&#xFF0C;&#x8FD9;&#x79CD;&#x5F62;&#x5F0F;&#x7684;&#x6539;&#x6027;&#x80DE;&#x5627;&#x5576;&#x7684;&#x4F5C;&#x7528;&#x4ECD;&#x7136;&#x4E0D;&#x660E;&#x6717;&#x3002;&#x5728;&#x8FD9;&#x91CC;&#x6211;&#x4EEC;&#x663E;&#x793A;&#x8BE5; 5 caC &#x662F; immunochemically &#x63A2;&#x6D4B;&#x8788;&#x5375;&#x5DE2;&#x4F53;&#x7EC6;&#x80DE;&#x4E2D;&#x3002;&#x6211;&#x4EEC;&#x8BC1;&#x660E; 5 hmC &#x548C; 5 caC &#x5DF2;&#x672C;&#x5730;&#x5316;&#x5230;&#x7C7B;&#x4F3C;&#x7684;&#x7A7A;&#x95F4;&#x5206;&#x5E03;&#x6A21;&#x5F0F;&#x4E0E;&#x8788;&#x6EE4;&#x6CE1;&#x7EC6;&#x80DE;&#x7684;&#x7EC6;&#x80DE;&#x6838;&#x67D3;&#x8272;&#x3002;&#x6211;&#x4EEC;&#x7684;&#x7814;&#x7A76;&#x7ED3;&#x679C;&#x8868;&#x660E;&#x8FD9; 5 carboxylcytosine &#x53EF;&#x80FD;&#x5728;&#x67D0;&#x4E9B;&#x751F;&#x7269;&#x7684;&#x4E0A;&#x4E0B;&#x6587;&#x4E2D;&#x53D1;&#x6325;&#x72EC;&#x7279;&#x7684;&#x529F;&#x80FD;&#x4F5C;&#x7528;&#x3002;

5 Carboxylcytosine &#x4EC5;&#x5C40;&#x9650;&#x4E8E; euchromatic &#x5730;&#x533A;&#x8788;&#x5375;&#x5DE2;&#x4E2D;&#x5375;&#x6CE1;&#x7EC6;&#x80DE;&#x7684;&#x7EC6;&#x80DE;&#x6838;&#x4E2D;&#x3002;

5-Methylcytosine (5-mC) is an epigenetic modification associated with gene repression. Recent studies demonstrated that 5-mC can be enzymatically oxidised into 5-hydroxymethylcytosine and further into 5-formylcytosine (5-fC) and 5-carboxylcytsine (5-caC). 5-caC has been found in embryonic stem cells and in mouse pre-implantation embryos but no detectable levels of this modification have been reported for somatic tissues to date. Whereas it has been suggested that 5-caC can serve as an intermediate in the process of active demethylation, the function of this form of modified cytosine remains obscure. Here we show that 5-caC is immunochemically detectable in somatic cells of axolotl ovary. We demonstrate that both 5-hmC and 5-caC are localized to the euchromatin in the nuclei of axolotl follicular cells with similar patterns of spatial distribution. Our results suggest that 5-carboxylcytosine may play a distinct functional role in certain biological contexts.

5-Carboxylcytosine is localized to euchromatic regions in the nuclei of follicular cells in axolotl ovary.

5-M&eacute;thylcytosine (5-mC) est une modification &eacute;pig&eacute;n&eacute;tique associ&eacute;e de r&eacute;pression g&eacute;nique. &Eacute;tudes r&eacute;centes ont d&eacute;montr&eacute; que mC-5 peut &ecirc;tre enzymatiquement oxyd&eacute; dans 5-hydroxym&eacute;thylcytosine et plus loin dans le 5-formylcytosine (5-fC) et 5-carboxylcytsine (5-caC). 5-ACC a &eacute;t&eacute; trouv&eacute; dans les cellules souches embryonnaires et chez les embryons pr&eacute;implantatoires, mais aucun des niveaux d&eacute;tectables de cette modification n&#39;ont &eacute;t&eacute; rapport&eacute;s pour les tissus somatiques &agrave; ce jour. Consid&eacute;rant qu&#39;il a &eacute;t&eacute; sugg&eacute;r&eacute; que 5-caC peut servir d&#39;interm&eacute;diaire dans le processus de d&eacute;m&eacute;thylation active, la fonction de cette forme de cytosine mis &agrave; jour le reste obscure. Nous montrons ici que 5-caC est immunochimiquement d&eacute;tectable dans les cellules somatiques de l&#39;ovaire de l&#39;axolotl. Nous d&eacute;montrons que les hmC-5 et 5-caC sont localis&eacute;es &agrave; l&#39;euchromatine dans les noyaux des cellules folliculaires de l&#39;axolotl avec des profils similaires de r&eacute;partition spatiale. Nos r&eacute;sultats sugg&egrave;rent que 5-carboxylcytosine peut jouer un r&ocirc;le fonctionnel distinct dans certains contextes biologiques.

5-Carboxylcytosine est localis&eacute;e dans des r&eacute;gions euchromatiques dans les noyaux des cellules folliculaires de l&#39;ovaire de l&#39;axolotl.

5-Methylcytosine (5-mC) ist eine epigenetische Modifikation gen Unterdr&uuml;ckung zugeordnet. J&uuml;ngste Studien gezeigt, dass 5-mC enzymatisch oxidiert in 5-Hydroxymethylcytosine und weiter in 5-Formylcytosine (5-fC) und 5-Carboxylcytsine (5-CaC) sein kann. 5-CaC in embryonale Stammzellen und Maus Pr&auml;implantations Embryonen gefunden wurde, aber keine nachweisbaren Niveaus dieser &Auml;nderung wurden bisher f&uuml;r somatische Gewebe beschrieben. W&auml;hrend es vorgeschlagen hat, dass diese 5-CaC als Zwischenprodukt in den Prozess der aktiven Demethylierung dienen kann, bleibt die Funktion dieser Form der modifizierten Cytosin obskuren. Hier zeigen wir, dass diese 5-CaC an somatischen Zellen der Eierst&ouml;cke Axolotl immunochemically nachweisbar ist. Wir zeigen, dass 5-HmC und 5-CaC an das Euchromatin in den Kernen der Axolotl follikul&auml;ren Zellen mit &auml;hnlichen Mustern der r&auml;umlichen Verteilung lokalisiert werden. Unsere Ergebnisse legen nahe, dass dieser 5-Carboxylcytosine in einem bestimmten biologischen Kontext eine unterschiedliche funktionale Rolle spielen kann.

5-Carboxylcytosine ist Histons Regionen in den Kernen der follikul&auml;ren Zellen in Axolotl Eierstock lokalisiert.

5-Metilcitosina (5-mC) &egrave; una modificazione epigenetica associata repressione genica. Recenti studi hanno dimostrato che 5-mC pu&ograve; essere enzimaticamente ossidato in 5-idrossimetilcitosina e ulteriormente in 5-formylcytosine (5-fC) e 5-carboxylcytsine (5-caC). 5-caC &egrave; stato trovato nelle cellule staminali embrionali e in preimpianto embrioni del mouse ma livelli rilevabili di questa modifica non sono stati segnalati per tessuti somatici per Data. Considerando che &egrave; stato suggerito che 5-caC pu&ograve; servire come un intermedio nel processo di demetilazione attiva, la funzione di questa forma di citosina modificato rimane oscura. Qui vi mostriamo che quello 5-caC &egrave; immunochemically rilevabile nelle cellule somatiche dell&#39;ovaia axolotl. Dimostriamo che sia hmC-5 e 5-caC sono localizzate a euchromatin nei nuclei delle cellule follicolari axolotl con simili modelli di distribuzione spaziale. I nostri risultati suggeriscono che 5-carboxylcytosine possa avere un ruolo funzionale distinto in determinati contesti biologici.

5-Carboxylcytosine &egrave; localizzata alle regioni eucromatina nei nuclei delle cellule follicolari nell&#39;ovaio axolotl.

5-&#x30E1;&#x30C1;&#x30EB;&#x30B7;&#x30C8;&#x30B7;&#x30F3; (5-mC) &#x907A;&#x4F1D;&#x5B50;&#x767A;&#x73FE;&#x6291;&#x5236;&#x3068;&#x95A2;&#x9023;&#x4ED8;&#x3051;&#x3089;&#x308C;&#x3066;&#x3044;&#x308B;&#x3001;&#x30A8;&#x30D4;&#x30B8;&#x30A7;&#x30CD;&#x30C6;&#x30A3;&#x30C3;&#x30AF;&#x306A;&#x4FEE;&#x98FE;&#x3067;&#x3059;&#x3002;&#x6700;&#x8FD1;&#x306E;&#x8ABF;&#x67FB;&#x306F;&#x305D;&#x306E; 5 mC &#x9175;&#x7D20;&#x306B;&#x3088;&#x3063;&#x3066; 5-&#x30D2;&#x30C9;&#x30ED;&#x30AD;&#x30B7;&#x30E1;&#x30C1;&#x30EB;&#x30B7;&#x30C8;&#x30B7;&#x30F3;&#x306B;&#x9178;&#x5316;&#x304A;&#x3088;&#x3073; 5-formylcytosine &#xFF08;5-fC&#xFF09; &#x3068; 5-carboxylcytsine &#xFF08;5-caC) &#x306B;&#x3055;&#x3089;&#x306B;&#x3059;&#x308B;&#x3053;&#x3068;&#x304C;&#x3067;&#x304D;&#x307E;&#x3059;&#x3092;&#x793A;&#x3057;&#x305F;&#x3002;5 caC &#x306F;&#x3001;&#x80DA;&#x6027;&#x5E79;&#x7D30;&#x80DE;&#x3068;&#x30DE;&#x30A6;&#x30B9;&#x7740;&#x5E8A;&#x524D;&#x80DA;&#x767A;&#x898B;&#x3055;&#x308C;&#x3066;&#x3044;&#x308B;&#x304C;&#x3001;&#x3053;&#x306E;&#x5909;&#x66F4;&#x306E;&#x691C;&#x51FA;&#x30EC;&#x30D9;&#x30EB;&#x4F53;&#x7D30;&#x80DE;&#x7D44;&#x7E54;&#x306E;&#x65E5;&#x4ED8;&#x306B;&#x5831;&#x544A;&#x3055;&#x308C;&#x3066;&#x3044;&#x307E;&#x305B;&#x3093;&#x3002;&#x305D;&#x306E; 5 caC &#x30A2;&#x30AF;&#x30C6;&#x30A3;&#x30D6;&#x8131;&#x30E1;&#x30C1;&#x30EB;&#x5316;&#x306E;&#x904E;&#x7A0B;&#x3067;&#x4E2D;&#x9593;&#x4F53;&#x3068;&#x3057;&#x3066;&#x4F7F;&#x7528;&#x3067;&#x304D;&#x308B;&#x304C;&#x793A;&#x5506;&#x3055;&#x308C;&#x3066;&#x3044;&#x308B;&#x306B;&#x5BFE;&#x3057;&#x3001;&#x5909;&#x66F4;&#x30B7;&#x30C8;&#x30B7;&#x30F3;&#x306E;&#x3053;&#x306E;&#x5F62;&#x5F0F;&#x306E;&#x95A2;&#x6570;&#x306F;&#x3042;&#x3044;&#x307E;&#x3044;&#x306A;&#x307E;&#x307E;&#x306B;&#x306A;&#x308A;&#x307E;&#x3059;&#x3002;&#x3053;&#x3053;&#x3067;&#x305D;&#x306E; 5 caC &#x304C;&#x4E21;&#x30A2;&#x30DB;&#x30ED;&#x30FC;&#x30C8;&#x30EB;&#x5375;&#x5DE3;&#x306E;&#x4F53;&#x7D30;&#x80DE;&#x306E;&#x691C;&#x51FA;&#x3067;&#x3042;&#x308B;&#x3092;&#x793A;&#x3057;&#x307E;&#x3059;&#x3002;&#x6211;&#x3005; 5 hmC &#x3068; 5 caC &#x30E6;&#x30FC;&#x30AF;&#x30ED;&#x30DE;&#x30C1;&#x30F3; &#x30A2;&#x30DB;&#x30ED;&#x30FC;&#x30C8;&#x30EB;&#x6FFE;&#x80DE;&#x7D30;&#x80DE;&#x306E;&#x6838;&#x306E;&#x7A7A;&#x9593;&#x5206;&#x5E03;&#x306E;&#x985E;&#x4F3C;&#x30D1;&#x30BF;&#x30FC;&#x30F3;&#x306B;&#x30ED;&#x30FC;&#x30AB;&#x30E9;&#x30A4;&#x30BA;&#x3092;&#x30C7;&#x30E2;&#x30F3;&#x30B9;&#x30C8;&#x30EC;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x3057;&#x307E;&#x3059;&#x3002;&#x305D;&#x306E; 5 carboxylcytosine &#x7279;&#x5B9A;&#x751F;&#x7269;&#x306E;&#x30B3;&#x30F3;&#x30C6;&#x30AD;&#x30B9;&#x30C8;&#x3067;&#x660E;&#x78BA;&#x306A;&#x6A5F;&#x80FD;&#x7684;&#x5F79;&#x5272;&#x3092;&#x518D;&#x751F;&#x53EF;&#x80FD;&#x6027;&#x304C;&#x3042;&#x308A;&#x307E;&#x3059;&#x304C;&#x793A;&#x5506;&#x3055;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;

5 Carboxylcytosine &#x30E6;&#x30FC;&#x30AF;&#x30ED;&#x30DE;&#x30C1;&#x30F3;&#x9818;&#x57DF;&#x30A2;&#x30DB;&#x30ED;&#x30FC;&#x30C8;&#x30EB;&#x5375;&#x5DE3;&#x306B;&#x304A;&#x3051;&#x308B;&#x5375;&#x80DE;&#x7D30;&#x80DE;&#x306E;&#x6838;&#x306B;&#x30ED;&#x30FC;&#x30AB;&#x30E9;&#x30A4;&#x30BA;&#x3055;&#x308C;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;

5-Methylcytosine (5-&#xC5E0;&#xC528;) &#xAD00;&#xB828; &#xC720;&#xC804;&#xC790; &#xC5B5;&#xC81C; &#xB41C; epigenetic &#xC218;&#xC815;&#xC785;&#xB2C8;&#xB2E4;. &#xCD5C;&#xADFC; &#xD559;&#xBB38;&#xC740; &#xADF8; 5 mC enzymatically 5 hydroxymethylcytosine&#xC5D0; &#xC2DC;&#xACC4;&#xC640; 5-formylcytosine (5-fC)&#xC640; 5-carboxylcytsine (5-caC)&#xC5D0; &#xCD94;&#xAC00; &#xB420; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4; &#xC124;&#xBA85; &#xD588;&#xB2E4;. 5-caC &#xB9C8;&#xC6B0;&#xC2A4; pre-implantation &#xBC30;&#xC544; &#xBC0F; &#xBC30;&#xC544; &#xC904;&#xAE30; &#xC138;&#xD3EC;&#xC5D0;&#xC11C; &#xBC1C;&#xACAC; &#xB418;&#xC5C8;&#xC2B5;&#xB2C8;&#xB2E4; &#xC788;&#xC9C0;&#xB9CC;&#xC774; &#xC218;&#xC815; &#xC5C6;&#xC74C; &#xAC10;&#xC9C0; &#xB808;&#xBCA8;&#xAE4C;&#xC9C0; &#xCCB4;&#xC138;&#xD3EC; &#xC870;&#xC9C1;&#xC5D0; &#xB300; &#xD55C; &#xBCF4;&#xACE0; &#xB418;&#xC5C8;&#xC2B5;&#xB2C8;&#xB2E4;. &#xADF8; 5-caC &#xD65C;&#xC131; demethylation &#xACFC;&#xC815; &#xC911;&#xAC04;&#xC774; &#xB420; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4; &#xC81C;&#xC548; &#xB418;&#xC5C8;&#xC2B5;&#xB2C8;&#xB2E4;, &#xBC18;&#xBA74;&#xC774; &#xD615;&#xD0DC;&#xC758; &#xC218;&#xC815;&#xB41C; &#xC2DC; &#xD568;&#xC218; &#xC560;&#xB9E4; &#xD55C; &#xB0A8;&#xC544; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC5EC;&#xAE30; &#xC6B0;&#xB9AC;&#xAC00; &#xADF8; 5 Cac&#xB294; immunochemically axolotl &#xB09C;&#xC18C;&#xC758; &#xCCB4;&#xC138;&#xD3EC;&#xC5D0;&#xC11C; &#xAC10;&#xC9C0; &#xBCF4;&#xC5EC;&#xC90D;&#xB2C8;&#xB2E4;. 5 HmC &#xBC0F; 5-caC &#xACF5;&#xAC04; &#xBD84;&#xD3EC;&#xC758; &#xBE44;&#xC2B7;&#xD55C; &#xD328;&#xD134;&#xC744; &#xAC00;&#xC9C4; axolotl follicular &#xC138;&#xD3EC;&#xC758; &#xD575;&#xC5D0; &#xC788;&#xB294; euchromatin&#xC5D0; &#xC9C0;&#xC5ED;&#xD654; &#xB429;&#xB2C8;&#xB2E4; &#xBCF4;&#xC5EC; &#xC90D;&#xB2C8;&#xB2E4;. &#xC6B0;&#xB9AC;&#xC758; &#xACB0;&#xACFC; 5-carboxylcytosine &#xC0DD;&#xBB3C; &#xD559;&#xC801; &#xD2B9;&#xC815; &#xCEE8;&#xD14D;&#xC2A4;&#xD2B8;&#xC5D0;&#xC11C; &#xBCC4;&#xAC1C; &#xAE30;&#xB2A5; &#xC5ED;&#xD560;&#xC744; &#xD560; &#xC218; &#xAC83;&#xC774; &#xC88B;&#xC2B5;&#xB2C8;&#xB2E4;.

5 Carboxylcytosine axolotl &#xB09C;&#xC18C;&#xC5D0; &#xC18C; &#xB0AD; &#xBAA8;&#xC591; &#xC138;&#xD3EC;&#xC758; &#xD575;&#xC5D0; &#xC788;&#xB294; euchromatic &#xC9C0;&#xC5ED;&#xC73C;&#xB85C; &#xC9C0;&#xC5ED;&#xD654; &#xB429;&#xB2C8;&#xB2E4;.

5-METILCITOSINA (5-mC) es una modificaci&oacute;n epigen&eacute;tica asociado con represi&oacute;n g&eacute;nica. Estudios recientes demostraron que 5-mC puede ser enzim&aacute;ticamente oxidado en 5-hidroximetilcitosina y m&aacute;s en 5-formylcytosine (5-fC) y 5-carboxylcytsine (5-caC). 5-caC se ha encontrado en las c&eacute;lulas madre embrionarias y en embriones de preimplantaci&oacute;n de rat&oacute;n pero no niveles detectables de esta modificaci&oacute;n se han divulgado para tejidos som&aacute;ticos hasta la fecha. Considerando que se ha sugerido que 5-caC puede servir como intermediario en el proceso de desmetilaci&oacute;n activa, la funci&oacute;n de esta forma de citosina modificado sigue siendo obscura. A continuaci&oacute;n os mostramos que eso 5-caC es inmunoqu&iacute;mica detectable en las c&eacute;lulas som&aacute;ticas del ovario de ajolote. Demostramos que tanto hmC-5 y 5-caC se localizan a la eucromatina en los n&uacute;cleos de las c&eacute;lulas foliculares de ajolote con similares patrones de distribuci&oacute;n espacial. Nuestros resultados sugieren que 5-carboxylcytosine puede jugar un papel funcional distinto en ciertos contextos biol&oacute;gicos.

5-Carboxylcytosine se localiza a regiones euchromatic en los n&uacute;cleos de las c&eacute;lulas foliculares en el ovario de ajolote.

5-Methylcytosine (5mC) is an epigenetic modification involved in regulation of gene activity during differentiation. Tet dioxygenases oxidize 5mC to 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC), and 5-carboxylcytosine (5caC). Both 5fC and 5caC can be excised from DNA by thymine-DNA glycosylase (TDG) followed by regeneration of unmodified cytosine via the base excision repair pathway. Despite evidence that this mechanism is operative in embryonic stem cells, the role of TDG-dependent demethylation in differentiation and development is currently unclear. Here, we demonstrate that widespread oxidation of 5hmC to 5caC occurs in postimplantation mouse embryos. We show that 5fC and 5caC are transiently accumulated during lineage specification of neural stem cells (NSCs) in culture and in vivo. Moreover, 5caC is enriched at the cell-type-specific promoters during differentiation of NSCs, and TDG knockdown leads to increased 5fC/5caC levels in differentiating NSCs. Our data suggest that active demethylation contributes to epigenetic reprogramming determining lineage specification in embryonic brain.

Transient accumulation of 5-carboxylcytosine indicates involvement of active demethylation in lineage specification of neural stem cells.

DNA methylation (5-methylcytosine (5mC)) patterns are often altered in cancers. Ten-eleven translocation (Tet) proteins oxidise 5mC to 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC) and 5-carboxylcytosine (5caC). In addition to their presumptive specific biological roles, these oxidised forms of 5mC may serve as intermediates in demethylation process. According to several reports, 5hmC levels are strongly decreased in cancers; however, the distribution of 5fC and 5caC in malignant tissue has not been studied.

ArticleTotal : 2	Year
Publication title Cited by 8	2016
Publication title Cited by 8	2017

Article	Year
5-Carboxylcytosine is localized to euchromatic regions in the nuclei of follicular cells in axolotl ovary. Nucleus (Austin, Tex.)\| PubMed ID: 23138778	2012
Transient accumulation of 5-carboxylcytosine indicates involvement of active demethylation in lineage specification of neural stem cells. Cell reports\| PubMed ID: 24882006	2014
5-Carboxylcytosine levels are elevated in human breast cancers and gliomas. Clinical epigenetics\| PubMed ID: 26300993	2015

Abdulkadir Abakir

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