Identification of OTX1 and OTX2 As Two Possible Molecular Markers for Sinonasal Carcinomas and Olfactory Neuroblastomas

OTX homeobox (HB) genes are expressed during embryonic morphogenesis and during the development of olfactory epithelium in adult organisms. Mutations ...

University of Insubria

In this report, we describe a polyacrylamide gel electrophoresis for the analysis of fluorophore-labelled hyaluronan (HA) and chondroitin sulfate (CS) Delta-disaccharides. The method utilizes derivatization of reducing ends of hyaluronan and the variously sulfated chondroitin sulfate Delta-disaccharides with 2-aminoacridone (AMAC), followed by electrophoresis in Tris-HCl buffer (pH 8.8), and polyacrylamide gel (T 25%/C 3.75%). The method was applied to the analysis of GAGs secreted into the culture medium of human aortic smooth muscle cells and the obtained results were compared with those analysed by fluorophore assisted carbohydrate electrophoresis (FACE). The described method is a useful and sensitive tool for the rapid monitoring of GAGs in high number of biologic samples.

Analysis of fluorophore-labelled hyaluronan and chondroitin sulfate disaccharides in biological samples.

This report describes a new formulation of polyacrylamide gel electrophoresis of fluorophore-labeled saccharides (PAGEFS) for the analysis of hyaluronan (HA) and chondroitin sulfate (CS) Delta-disaccharides. PAGEFS relies on derivatization of reducing ends of HA- and the variously sulfated CS-derived Delta-disaccharides with 2-aminoacridone (AMAC), followed by electrophoresis under optimized buffer conditions (Tris-borate and Tris-HCl) and on polyacrylamide gels (25% T/3.75% C). The method was applied to the analysis of glycosaminoglycans (GAGs) from the human umbilical cord tissue and GAGs isolated from human aortic smooth muscle cell cultures. The obtained results were in agreement with those obtained after an analysis with high-performance liquid chromatography (HPLC). On the basis of these results, PAGEFS is a rapid and sensitive method for the analysis of the total amount of HA- and CS-derived disaccharides, as it allows analyzing 20 samples in minigels in one run and provides quantitation with relatively high sensitivity (less than 25 pmol per disaccharide). In addition, PAGEFS overcomes the lack of commercial gels described previously for the separation of AMAC-labeled disaccharides. Therefore, the method proposed here is an economic and useful tool for a fast screening of GAGs in biological samples, particularly when a high number of samples should be analyzed.

Polyacrylamide gel electrophoresis of fluorophore-labeled hyaluronan and chondroitin sulfate disaccharides: application to the analysis in cells and tissues.

Single nucleotide polymorphisms (SNPs) are becoming the most common type of markers used in genetic analysis. In the present report a SNP has been chosen to test the applicability of Real Time PCR to discriminate and quantify SNPs alleles on DNA pools. Amplification Refractory Mutation System (ARMS) and Mismatch Amplification Mutation Assay (MAMA) has been applied. Each assay has been pre-validated testing specificity and performances (linearity, PCR efficiency, interference limit, limit of detection, limit of quantification, precision and accuracy). Both the approaches achieve a precise and accurate estimation of the allele frequencies on pooled DNA samples in the range from 5 % to 95 % and don't require standard curves or calibrators. The lowest measurement that could be significantly distinguished from the background noise has been determined around the 1 % for both the approaches, allowing to extend the range of quantifications from 1 % to 99 %. Furthermore applicability of Real Time PCR assays for general diagnostic purposes is discussed.

Different real time PCR approaches for the fine quantification of SNP's alleles in DNA pools: assays development, characterization and pre-validation.

The evaluation of residual disease, which has prognostic value in the treatment of hematological malignancies, is currently assessed by scoring a limited number of cells by karyotyping and molecular cytogenetics. Real-time polymerase chain reaction (PCR) is an easier and more sensitive technique, enables analysis of a larger number of cells, and decreases sampling error. However, real-time PCR has been applied only to target transcripts of fusion genes. Here, we considered two real-time PCR strategies to quantify a number of cells carrying a partial deletion of chromosome 7 mixed with normal disomic cells. The first strategy was based on the amplification of two sequences, one on chromosome 7 and the other on chromosome 14. In the second strategy residual disease was assessed by the ratio between the two alleles of a bi-allelic marker, mapped on chromosome 7, measured with allele-specific assays. Precision and accuracy of the two approaches were tested by reference samples with nominal values of residual disease ranging from 2 to 95%. As expected the second strategy resulted in more precise and accurate monitoring within the range from 5 to 95%. Furthermore, this method may be applied to assess the number of dysplastic or neoplastic clones carrying any unbalanced chromosome changes.

Establishment and study of different real-time polymerase chain reaction assays for the quantification of cells with deletions of chromosome 7.

&#x4E73;&#x817A;&#x72EC;&#x7279;&#x7684;&#x673A;&#x6784;&#x7684;&#x4E3B;&#x8981;&#x5F62;&#x5F0F;&#xFF0C;&#x9752;&#x6625;&#x671F;&#x662F;&#x4E00;&#x4E2A;&#x7406;&#x60F3;&#x7684;&#x6A21;&#x578B;&#xFF0C;&#x4EE5;&#x7814;&#x7A76;&#x5728;&#x53D1;&#x5C55;&#x548C;&#x80BF;&#x7624;&#x53D1;&#x751F; (HB) &#x540C;&#x6E90;&#x76D2;&#x57FA;&#x56E0;&#x7684;&#x529F;&#x80FD;&#x3002;HB &#x57FA;&#x56E0;&#x7EC4;&#x6210;&#x4E00;&#x4E2A;&#x5927;&#x5BB6;&#x5EAD;&#x7684;&#x53D1;&#x5C55;&#x5177;&#x6709;&#x5173;&#x952E;&#x4F5C;&#x7528;&#xFF0C;&#x7EC6;&#x80DE;&#x751F;&#x957F;&#x548C;&#x5206;&#x5316;&#x7684;&#x76D1;&#x7BA1;&#x3002;&#x5728;&#x6B63;&#x5E38;&#x7684;&#x4E73;&#x817A;&#x7EC4;&#x7EC7;&#xFF0C;&#x540C;&#x6E90;&#x76D2;&#x57FA;&#x56E0;&#x53C2;&#x4E0E;&#x5BFC;&#x7BA1;&#x5F62;&#x6210;&#x3001; &#x4E0A;&#x76AE;&#x5206;&#x652F;&#x548C; lobulo &#x80BA;&#x6CE1;&#x53D1;&#x5C55;&#x7684;&#x89C4;&#x7BA1;&#x4E0A;&#x76AE;&#x7EC6;&#x80DE;&#x589E;&#x6B96;&#x548C;&#x5206;&#x5316;&#x3002;HB &#x57FA;&#x56E0;&#x63A7;&#x5236;&#x57FA;&#x8D28;&#x7EC6;&#x80DE;&#x548C;&#x4E0A;&#x76AE;&#x7EC6;&#x80DE;&#x4E2D;&#x7684;&#x7A7A;&#x95F4;&#x548C;&#x65F6;&#x95F4;&#x7684;&#x65B9;&#x5F0F;&#x3002;&#x4ED6;&#x4EEC;&#x534F;&#x8C03;&#x53D7;&#x6FC0;&#x7D20;&#x548C;&#x7EC6;&#x80DE;&#x5916;&#x57FA;&#x8D28;&#xFF0C;&#x8FD9;&#x8868;&#x660E;&#x8BB8;&#x591A;&#x4FE1;&#x53F7;&#x901A;&#x8DEF;&#x53C2;&#x4E0E;&#x4E86;&#x540C;&#x6E90;&#x76D2;&#x57FA;&#x56E0;&#x529F;&#x80FD;&#x3002;&#x540C;&#x6E90;&#x76D2;&#x57FA;&#x56E0; misexpressed &#x5728;&#x52A8;&#x7269;&#x6A21;&#x578B;&#x4E2D;&#xFF0C;&#x5F53;&#x5728;&#x4E73;&#x817A;&#x53D1;&#x80B2;&#x4E2D;&#x663E;&#x793A;&#x4E0D;&#x540C;&#x7684;&#x7F3A;&#x9677;&#x3002;&#x5F02;&#x5E38;&#x8868;&#x8FBE;&#x7684;&#x540C;&#x6E90;&#x76D2;&#x57FA;&#x56E0;&#xFF0C;&#x5916;&#x57DF;&#x6216;&#x7F16;&#x7801;&#xFF0C;&#x627E;&#x5230;&#x539F;&#x53D1;&#x764C;&#x548C;&#x4E73;&#x817A;&#x764C;&#x3002;Otx1 &#x8840;&#x7EA2;&#x86CB;&#x767D;&#x57FA;&#x56E0;&#x662F;&#x76D1;&#x7BA1;&#x795E;&#x7ECF;&#x7CFB;&#x7EDF;&#x53D1;&#x80B2;&#x80DA;&#x80CE;&#x53D1;&#x751F;&#x8FC7;&#x7A0B;&#x4E2D;&#x7684;&#x4E00;&#x4E2A;&#x7ECF;&#x5178;&#x3002;Otx1 &#x5728;&#x5782;&#x4F53;&#x524D;&#x7684;&#x8F6C;&#x5F55;&#x5E76;&#x5728;&#x6FC0;&#x6D3B;&#x7684;&#x5782;&#x4F53;&#x6FC0;&#x7D20;&#x3001; &#x8F6C;&#x5F55;&#x548C;&#x9020;&#x8840;&#x529F;&#x80FD;&#x3001; &#x589E;&#x5F3A;&#x7684;&#x591A;&#x80FD;&#x5E72;&#x7EC6;&#x80DE;&#x548C;&#x7EA2;&#x7CFB;&#x5206;&#x5316;&#x4E2D;&#x7684;&#x4F5C;&#x7528;&#x3002;Otx1 &#x4ECD;&#x5728;&#x7EA2;&#x7EC6;&#x80DE;&#x548C; megacaryocytic &#x6CBF;&#x88AD;&#x7684;&#x6210;&#x719F;&#x7EC6;&#x80DE;&#x4E2D;&#x68C0;&#x6D4B;&#x5230;&#x3002;&#x5468;&#x671F;&#x6027;&#x53D1;&#x5C55;&#x8FC7;&#x7A0B;&#x7684;&#x4E73;&#x817A;&#x7EC4;&#x7EC7;&#x4E2D;&#xFF0C;Otx1 &#x57FA;&#x56E0;&#x6297;&#x539F;&#x5728;&#x54FA;&#x4E73;&#x671F;&#xFF0C;&#x786E;&#x8BA4;&#x6B64;&#x8F6C;&#x5F55;&#x56E0;&#x5B50;&#x5728;&#x7EC6;&#x80DE;&#x5206;&#x5316;&#x4E2D;&#x7684;&#x4F5C;&#x7528;&#x3002;&#x6700;&#x8FD1;&#x7684;&#x7814;&#x7A76;&#x62A5;&#x544A; Otx1 &#x6297;&#x539F;&#x5728;&#x4E73;&#x817A;&#x764C;&#x3002;&#x5728;&#x80DA;&#x80CE;&#x53D1;&#x751F;&#xFF0C;&#x671F;&#x95F4;&#x8868;&#x793A;&#xFF0C;Otx1&#xFF0C;&#x5B83;&#x764C;&#x53D8;&#xFF0C;&#x8FD9;&#x610F;&#x5473;&#x7740;&#x5176;&#x53EF;&#x80FD;&#x529F;&#x80FD;&#x7684;&#x80BF;&#x7624;&#x7EC6;&#x80DE;&#x5206;&#x5316;&#x8FC7;&#x7A0B;&#x4E2D;&#x518D;&#x6B21;&#x8868;&#x793A;&#x3002;

&#x4E73;&#x817A;&#x7EC4;&#x7EC7;&#x548C; Otx1 &#x7684;&#x540C;&#x6E90;&#x76D2;&#x57FA;&#x56E0;&#x3002;

The mammary gland, the unique organ that primarily form at puberty, is an ideal model to study the functions of homeobox (HB) genes in both development and tumorigenesis. HB genes comprise a large family of developmental regulators that have a critical role in cell growth and differentiation. In the normal mammary gland, homeobox genes are involved in ductal formation, epithelial branching, and lobulo-alveolar development by regulating epithelial proliferation and differentiation. The HB genes are controlled in a spatial and temporal manner in both stromal and epithelial cells. They are coordinately regulated by hormones and extracellular matrix, suggesting that many signaling pathways are involved in homeobox gene functions. When homeobox genes are misexpressed in animal models, different defects are displayed in mammary gland development. Aberrant expression of homeobox genes, overexpressed or downregulated, is found in primary carcinomas and in breast cancer. The Otx1 HB gene is a classic regulatory of nervous system development during embryogenesis. Postnatally Otx1 is transcribed in the anterior pituitary gland, where activates transcription of the pituitary hormones, and plays a role in hematopoiesis, enhancing pluripotent cells, and erythroid differentiation. Otx1 can still be detected in mature cells of the erythroid and megacaryocytic lineage. During cyclical development of mammary gland, the Otx1 gene is overexpressed in lactation, confirming a role of this transcription factor in cell differentiation. Recent studies report that Otx1 is overexpressed in breast cancer. Otx1 is expressed during embryogenesis, and it is expressed again during carcinogenesis, implying its possible function in differentiation of neoplastic cells.

The mammary gland and the homeobox gene Otx1.

La glande mammaire, l&#39;unique organe qui forment principalement &agrave; la pubert&eacute;, est un mod&egrave;le id&eacute;al pour &eacute;tudier les fonctions des g&egrave;nes homeobox (HB) dans le d&eacute;veloppement et la tumorigen&egrave;se. G&egrave;nes HB forment une grande famille de r&eacute;gulateurs du d&eacute;veloppement qui ont un r&ocirc;le essentiel dans la croissance cellulaire et la diff&eacute;renciation. Dans la glande mammaire normale, homeobox g&egrave;nes sont impliqu&eacute;s dans la formation de ductal, ramification &eacute;pith&eacute;liales et d&eacute;veloppement lobulo-alv&eacute;olaire en r&eacute;gulant la diff&eacute;renciation et la prolif&eacute;ration &eacute;pith&eacute;liale. Les g&egrave;nes de HB sont contr&ocirc;l&eacute;s de fa&ccedil;on spatiale et temporelle en tant les cellules &eacute;pith&eacute;liales du stroma. Coordonner, ils sont r&eacute;glement&eacute;s par les hormones et la matrice extracellulaire, ce qui sugg&egrave;re que les nombreuses voies de signalisation sont impliqu&eacute;s dans les fonctions du g&egrave;ne hom&eacute;otique. Lorsque homeobox g&egrave;nes sont misexpressed dans des mod&egrave;les animaux, diff&eacute;rents d&eacute;fauts sont affich&eacute;es dans le d&eacute;veloppement de la glande mammaire. Expression aberrante de homeobox g&egrave;nes surexprim&eacute;s ou r&eacute;prim&eacute;s, se trouve dans les carcinomes primaires et dans le cancer du sein. Le g&egrave;ne Otx1 HB est un classique r&eacute;glementaire du d&eacute;veloppement du syst&egrave;me nerveux au cours de l&#39;embryogen&egrave;se. Apr&egrave;s la naissance Otx1 est transcrit dans l&#39;hypophyse ant&eacute;rieure, o&ugrave; active la transcription des hormones hypophysaires, et joue un r&ocirc;le dans l&#39;h&eacute;matopo&iuml;&egrave;se, renforcement des cellules pluripotentes et la diff&eacute;renciation &eacute;rythro&iuml;de. Otx1 peut encore &ecirc;tre d&eacute;tect&eacute; dans les cellules matures de la lign&eacute;e &eacute;rythro&iuml;de et megacaryocytic. Au cours du d&eacute;veloppement cyclique de la glande mammaire, le g&egrave;ne Otx1 est surexprim&eacute; dans la lactation, confirmant le r&ocirc;le de ce facteur de transcription dans la diff&eacute;renciation cellulaire. Des &eacute;tudes r&eacute;centes rapportent que Otx1 est surexprim&eacute;e dans le cancer du sein. Otx1 est exprim&eacute;e au cours de l&#39;embryogen&egrave;se, et il s&#39;exprime encore une fois au cours de la canc&eacute;rogen&egrave;se, ce qui implique son r&ocirc;le possible dans la diff&eacute;renciation des cellules n&eacute;oplasiques.

La glande mammaire et le g&egrave;ne hom&eacute;otique Otx1.

Der Brustdr&uuml;se, die einzigartige Orgel, die vor allem in der Pubert&auml;t, zu bilden ist ein ideales Modell, um die Funktionen der (HB) Hox-Gene sowohl die Entwicklung als auch die Tumorentstehung zu studieren. HB-Gene umfassen eine gro&szlig;e Familie von Entwicklungsst&ouml;rungen Regulatoren, die eine entscheidende Rolle im Zellwachstum und Differenzierung. In der normalen Brustdr&uuml;se sind Hox-Gene in duktale Formation, epitheliale verzweigen und Lobulo-alveol&auml;re Entwicklung beteiligt, durch die Regulierung epitheliale Proliferation und Differenzierung. Die HB-Gene werden in einer r&auml;umlichen und zeitlichen Weise in sowohl stromal und epitheliale Zellen gesteuert. Sie sind coordinately geregelt durch Hormone und extrazellul&auml;re Matrix, was darauf hindeutet, dass viele Signalwege in Homeobox Genfunktionen beteiligt sind. Wenn Hox-Gene in Tiermodellen misexpressed sind, werden verschiedene M&auml;ngel in der Brustdr&uuml;se Entwicklung angezeigt. Aberrante Expression von Hox-Gene, Pleuramesotheliom oder Downregulated, befinden sich in prim&auml;re Karzinome sowie bei Brustkrebs. Das Otx1 HB-gen ist ein Klassiker, der Regulierung des Nervensystems Entwicklung w&auml;hrend der Embryogenese. Postnatal ist Otx1 in den vorderen Hypophyse, transkribiert wo Transkription der Hypophyse Hormone aktiviert, und spielt eine Rolle bei der Blutbildung, Verbesserung pluripotenten Zellen und Erythroid Differenzierung. Otx1 k&ouml;nnen noch im Reifen Zellen der Erythroid und Megacaryocytic Abstammung nachgewiesen werden. W&auml;hrend der zyklischen Entwicklung der Brustdr&uuml;se ist das Otx1-gen in der Stillzeit, best&auml;tigt eine Rolle dieser Transkriptionsfaktor in Zelldifferenzierung Pleuramesotheliom. Neuere Studien berichten, dass Otx1 bei Brustkrebs Pleuramesotheliom ist. Otx1 wird w&auml;hrend der Embryogenese exprimiert, und wieder w&auml;hrend der Karzinogenese, was seine m&ouml;gliche Funktion in der Differenzierung der neoplastischen Zellen ausgedr&uuml;ckt wird.

Der Brustdr&uuml;se und der Hom&ouml;obox-gen Otx1.

La ghiandola mammaria, l&#39;unico organo che formano principalmente durante la pubert&agrave;, &egrave; un modello ideale per studiare le funzioni dei geni homeobox (HB) lo sviluppo e la tumorigenesi. Geni HB costituiscono una grande famiglia di regolatori dello sviluppo che hanno un ruolo critico nella crescita cellulare e la differenziazione. Nella ghiandola mammaria normale, homeobox geni sono coinvolti nella formazione duttale, ramificazione epiteliale e sviluppo lobulo-alveolare regolando la differenziazione e la proliferazione epiteliale. I geni di HB sono controllati in modo spaziale e temporale in cellule stromali ed epiteliali. Coordinately sono regolati da ormoni e matrice extracellulare, suggerendo che nelle funzioni del gene homeobox sono coinvolte molte vie di segnalazione. Quando i geni dell&#39;homeobox sono misexpressed in modelli animali, vengono visualizzati diversi difetti nello sviluppo della ghiandola mammaria. Aberrante espressione di geni homeobox, overexpressed o downregolata, &egrave; trovato in carcinomi primari e nel cancro al seno. Il gene Otx1 HB &egrave; un classico regolamentazione dello sviluppo del sistema nervoso durante l&#39;embriogenesi. Dopo la nascita &egrave; trascritto Otx1 nella ghiandola pituitaria anteriore, dove attiva la trascrizione degli ormoni ipofisari, e svolge un ruolo nella differenziazione eritroide ematopoiesi e migliorare le cellule pluripotenti. Otx1 pu&ograve; ancora essere rilevato in cellule mature del lignaggio eritroidi e megacaryocytic. Durante lo sviluppo ciclico della ghiandola mammaria, il gene Otx1 &egrave; overexpressed in lattazione, confermando un ruolo di questo fattore di trascrizione nel differenziamento cellulare. Studi recenti riportano che Otx1 &egrave; overexpressed nel cancro al seno. Otx1 &egrave; espresso durante l&#39;embriogenesi e si esprime ancora una volta durante la carcinogenesi, implicando la sua possibile funzione di differenziazione delle cellule neoplastiche.

La ghiandola mammaria e il homeobox gene Otx1.

&#x4E3B;&#x306B;&#x601D;&#x6625;&#x671F;&#x306B;&#x306F;&#x3001;&#x30D5;&#x30A9;&#x30FC;&#x30E0;&#x56FA;&#x6709;&#x306E;&#x81D3;&#x5668;&#x306E;&#x4E73;&#x817A;&#x306E;&#x958B;&#x767A;&#x3068;&#x816B;&#x760D;&#x767A;&#x751F;&#x306B;&#x304A;&#x3051;&#x308B;&#x30DB;&#x30E1;&#x30AA; &#x30DC;&#x30C3;&#x30AF;&#x30B9; (HB) &#x907A;&#x4F1D;&#x5B50;&#x6A5F;&#x80FD;&#x7814;&#x7A76;&#x306B;&#x7406;&#x60F3;&#x7684;&#x306A;&#x30E2;&#x30C7;&#x30EB;&#x3067;&#x3059;&#x3002;HB &#x907A;&#x4F1D;&#x5B50;&#x306F;&#x7D30;&#x80DE;&#x5897;&#x6B96;&#x3068;&#x5206;&#x5316;&#x306B;&#x91CD;&#x8981;&#x306A;&#x5F79;&#x5272;&#x304C;&#x3042;&#x308B;&#x767A;&#x9054;&#x306E;&#x30EC;&#x30AE;&#x30E5;&#x30EC;&#x30FC;&#x30BF;&#x306E;&#x5927;&#x5BB6;&#x65CF;&#x3092;&#x69CB;&#x6210;&#x3057;&#x307E;&#x3059;&#x3002;&#x6B63;&#x5E38;&#x4E73;&#x817A;&#x306E;&#x30DB;&#x30E1;&#x30AA; &#x30DC;&#x30C3;&#x30AF;&#x30B9;&#x907A;&#x4F1D;&#x5B50;&#x4E73;&#x7BA1;&#x5F62;&#x6210;, &#x4E0A;&#x76AE;&#x5206;&#x5C90;&#x3068; lobulo &#x6B6F;&#x69FD;&#x958B;&#x767A;&#x4E0A;&#x76AE;&#x306E;&#x5897;&#x6B96;&#x3068;&#x5206;&#x5316;&#x3092;&#x8ABF;&#x7BC0;&#x3059;&#x308B;&#x3053;&#x3068;&#x306B;&#x3088;&#x3063;&#x3066;&#x304B;&#x304B;&#x308F;&#x3063;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;HB &#x907A;&#x4F1D;&#x5B50;&#x9593;&#x8CEA;&#x7D30;&#x80DE;&#x306A;&#x3089;&#x3073;&#x306B;&#x4E0A;&#x76AE;&#x7D30;&#x80DE;&#x3067;&#x306E;&#x6642;&#x7A7A;&#x7684;&#x5236;&#x5FA1;&#x3055;&#x308C;&#x307E;&#x3059;&#x3002;&#x30DB;&#x30EB;&#x30E2;&#x30F3;&#x3068;&#x7D30;&#x80DE;&#x5916;&#x30DE;&#x30C8;&#x30EA;&#x30C3;&#x30AF;&#x30B9;&#x306B;&#x3088;&#x3063;&#x3066;&#x591A;&#x304F;&#x306E;&#x30B7;&#x30B0;&#x30CA;&#x30EB;&#x4F1D;&#x9054;&#x7D4C;&#x8DEF;&#x30DB;&#x30E1;&#x30AA; &#x30DC;&#x30C3;&#x30AF;&#x30B9;&#x907A;&#x4F1D;&#x5B50;&#x306E;&#x6A5F;&#x80FD;&#x306B;&#x95A2;&#x4E0E;&#x3057;&#x3066;&#x3044;&#x308B;&#x3053;&#x3068;&#x3092;&#x793A;&#x5506;&#x5F7C;&#x3089;&#x30EA;&#x30BA;&#x30DF;&#x30AB;&#x30EB;&#x898F;&#x5236;&#x3055;&#x308C;&#x307E;&#x3059;&#x3002;&#x30DB;&#x30E1;&#x30AA; &#x30DC;&#x30C3;&#x30AF;&#x30B9;&#x907A;&#x4F1D;&#x5B50;&#x304C;&#x52D5;&#x7269;&#x30E2;&#x30C7;&#x30EB;&#x3067; misexpressed &#x3044;&#x308B;&#x3068;&#x304D;&#x306B;&#x3001;&#x7570;&#x306A;&#x308B;&#x6B20;&#x9665;&#x4E73;&#x817A;&#x306E;&#x958B;&#x767A;&#x3067;&#x8868;&#x793A;&#x3055;&#x308C;&#x307E;&#x3059;&#x3002;&#x7570;&#x5E38;&#x767A;&#x73FE;&#x306E;&#x30DB;&#x30E1;&#x30AA; &#x30DC;&#x30C3;&#x30AF;&#x30B9;&#x907A;&#x4F1D;&#x5B50;&#x306E;&#x904E;&#x5270;&#x767A;&#x73FE;&#x307E;&#x305F;&#x306F; downregulated&#x3001;&#x30D7;&#x30E9;&#x30A4;&#x30DE;&#x30EA;&#x764C;&#x3068;&#x4E73;&#x764C;&#x3092;&#x767A;&#x898B;&#x3057;&#x307E;&#x3059;&#x3002;Otx1 HB &#x907A;&#x4F1D;&#x5B50;&#x30AF;&#x30E9;&#x30B7;&#x30C3;&#x30AF;&#x306A;&#x80DA;&#x5F62;&#x6210;&#x904E;&#x7A0B;&#x306B;&#x304A;&#x3051;&#x308B;&#x795E;&#x7D4C;&#x7CFB;&#x958B;&#x767A;&#x306E;&#x898F;&#x5236;&#x3067;&#x3059;&#x3002;&#x5C3F;&#x8DEF;&#x30C9;&#x30EC;&#x30CA;&#x30FC;&#x30B8; Otx1 &#x8133;&#x4E0B;&#x5782;&#x4F53;&#x524D;&#x8449;&#x3001;&#x8EE2;&#x5199;&#x3055;&#x308C;&#x308B;&#x5834;&#x6240;&#x3001;&#x4E0B;&#x5782;&#x4F53;&#x30DB;&#x30EB;&#x30E2;&#x30F3;&#x306E;&#x8EE2;&#x5199;&#x3092;&#x6D3B;&#x6027;&#x5316;&#x3057;&#x3001;&#x9020;&#x8840;&#x3001;&#x5F37;&#x5316;&#x591A;&#x80FD;&#x6027;&#x7D30;&#x80DE;&#x3001;&#x8D64;&#x8840;&#x7403;&#x5206;&#x5316;&#x306B;&#x304A;&#x3051;&#x308B;&#x5F79;&#x5272;&#x3092;&#x679C;&#x305F;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;Otx1 &#x306F;&#x3001;&#x307E;&#x3060;&#x8D64;&#x82BD;&#x7403;&#x304A;&#x3088;&#x3073; megacaryocytic &#x306E;&#x7CFB;&#x7D71;&#x306E;&#x6210;&#x719F;&#x7D30;&#x80DE;&#x3067;&#x691C;&#x51FA;&#x3067;&#x304D;&#x307E;&#x3059;&#x3002;&#x4E73;&#x817A;&#x306E;&#x5FAA;&#x74B0;&#x306E;&#x958B;&#x767A;&#x4E2D;&#x306F;&#x3001;Otx1 &#x907A;&#x4F1D;&#x5B50;&#x3092;&#x904E;&#x5270;&#x767A;&#x73FE;&#x6388;&#x4E73;&#x671F;&#x3001;&#x3053;&#x306E;&#x7D30;&#x80DE;&#x306E;&#x5206;&#x5316;&#x306B;&#x304A;&#x3051;&#x308B;&#x8EE2;&#x5199;&#x56E0;&#x5B50;&#x306E;&#x5F79;&#x5272;&#x3092;&#x78BA;&#x8A8D;&#x3057;&#x307E;&#x3059;&#x3002;&#x6700;&#x8FD1;&#x306E;&#x7814;&#x7A76;&#x306F;&#x3001;Otx1 &#x304C;&#x4E73;&#x764C;&#x3067;&#x904E;&#x5270;&#x306B;&#x767A;&#x73FE;&#x5831;&#x544A;&#x3057;&#x307E;&#x3059;&#x3002;Otx1 &#x80DA;&#x767A;&#x751F;&#x4E2D;&#x306B;&#x8868;&#x73FE;&#x3055;&#x308C;&#x3001;&#x518D;&#x3073;&#x767A;&#x304C;&#x3093;, &#x305D;&#x306E;&#x816B;&#x760D;&#x7D30;&#x80DE;&#x306E;&#x5206;&#x5316;&#x306B;&#x304A;&#x3051;&#x308B;&#x53EF;&#x80FD;&#x306A;&#x6A5F;&#x80FD;&#x3092;&#x610F;&#x5473;&#x3057;&#x3066;&#x4E2D;&#x3092;&#x8868;&#x660E;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;

&#x4E73;&#x817A;&#x3068;&#x30DB;&#x30E1;&#x30AA; &#x30DC;&#x30C3;&#x30AF;&#x30B9;&#x907A;&#x4F1D;&#x5B50; Otx1&#x3002;

&#xC720; &#xBC29; &#xC120;, &#xC8FC;&#xB85C; &#xC0AC;&#xCD98;&#xAE30;&#xC5D0; &#xD615;&#xC131; &#xD558;&#xB294; &#xB3C5;&#xD2B9;&#xD55C; &#xAE30;&#xAD00; &#xAC1C;&#xBC1C; &#xBC0F; tumorigenesis (HB) homeobox &#xC720;&#xC804;&#xC790;&#xC758; &#xAE30;&#xB2A5; &#xC5F0;&#xAD6C;&#xC5D0; &#xC774;&#xC0C1;&#xC801;&#xC778; &#xBAA8;&#xB378;&#xC785;&#xB2C8;&#xB2E4;. HB &#xC720;&#xC804;&#xC790; &#xC138;&#xD3EC; &#xC131;&#xC7A5;&#xACFC; &#xBD84;&#xD654;&#xC5D0; &#xC911;&#xC694; &#xD55C; &#xC5ED;&#xD560;&#xC744;&#xAC00;&#xC9C0;&#xACE0; &#xBC1C;&#xB2EC; &#xB808; &#xADE4; &#xB808;&#xC774; &#xD130;&#xC758; &#xD070; &#xAC00;&#xC871;&#xC73C;&#xB85C; &#xAD6C;&#xC131; &#xB429;&#xB2C8;&#xB2E4;. &#xC77C;&#xBC18; &#xC720;&#xC120; &#xC120; homeobox &#xC720;&#xC804;&#xC790;&#xB294; &#xC0C1;&#xD53C; &#xC99D;&#xC2DD;&#xACFC; &#xBD84;&#xD654;&#xB97C; &#xC870;&#xC808; &#xD558; &#xC5EC; ductal &#xD615;&#xC131;, &#xC0C1;&#xD53C; &#xBD84;&#xAE30; &#xBC0F; lobulo &#xCE58;&#xACBD; &#xAC1C;&#xBC1C;&#xC5D0;&#xC11C; &#xD3EC;&#xD568; &#xB41C;&#xB2E4;. HB &#xC720;&#xC804;&#xC790;&#xB294; &#xAE30;&#xC9C8;&#xACFC; &#xC0C1;&#xD53C; &#xC138;&#xD3EC;&#xC5D0;&#xC11C; &#xACF5;&#xAC04; &#xBC0F; &#xC2DC;&#xAC04; &#xBC29;&#xC2DD;&#xC73C;&#xB85C; &#xC81C;&#xC5B4; &#xB429;&#xB2C8;&#xB2E4;. &#xADF8;&#xB4E4;&#xC740; coordinately &#xC81C;&#xC548; &#xD558;&#xB294; &#xB9CE;&#xC740; &#xC2E0;&#xD638; &#xACBD;&#xB85C; homeobox &#xC720;&#xC804;&#xC790; &#xAE30;&#xB2A5;&#xC5D0; &#xAD00;&#xB828; &#xB41C; &#xD638;&#xB974;&#xBAAC;&#xACFC; &#xC138;&#xD3EC; &#xC678; &#xB9E4;&#xD2B8;&#xB9AD;&#xC2A4;&#xC5D0; &#xC758;&#xD574; &#xD1B5;&#xC81C; &#xB41C;&#xB2E4;. Homeobox &#xC720;&#xC804;&#xC790;&#xB294; &#xB3D9;&#xBB3C; &#xBAA8;&#xB378;&#xC5D0;&#xC11C; misexpressed&#xB294;, &#xC720;&#xC120; &#xBC1C;&#xB2EC;&#xC5D0; &#xB2E4;&#xB978; &#xACB0;&#xD568;&#xC774; &#xD45C;&#xC2DC; &#xB429;&#xB2C8;&#xB2E4;. Overexpressed homeobox &#xC720;&#xC804;&#xC790; &#xB610;&#xB294; downregulated, &#xD0C8; &#xC120; &#xC2DD; &#xC8FC; carcinomas &#xBC0F; &#xC720;&#xBC29;&#xC554; &#xBC1C;&#xACAC; &#xB41C;&#xB2E4;. Otx1 HB &#xC720;&#xC804;&#xC790; embryogenesis &#xC911; &#xC2E0; &#xC2DC;&#xC2A4;&#xD15C; &#xAC1C;&#xBC1C;&#xC758; &#xADDC;&#xC81C; &#xACE0;&#xC804; &#xC774;&#xB2E4;. Postnatally Otx1 &#xC55E;&#xCABD;&#xC5D0; &#xB1CC; &#xD558; &#xC218; &#xCCB4;&#xC5D0;&#xC11C; &#xBCC0;&#xD558;&#xAC8C; &#xB418; &#xC788;&#xB294; &#xB1CC; &#xD558; &#xC218; &#xCCB4; &#xD638;&#xB974;&#xBAAC;&#xC758; &#xC804;&#xC0AC;&#xB97C; &#xD65C;&#xC131;&#xD654; &#xD558; &#xACE0; &#xC870;, &#xAC15;&#xD654; pluripotent &#xC140; &#xBC0F; erythroid &#xCC28;&#xBCC4;&#xD654;&#xC5D0; &#xC5ED;&#xD560;&#xC744; &#xD55C;&#xB2E4;. Otx1 erythroid &#xBC0F; megacaryocytic &#xD608;&#xD1B5;&#xC758; &#xC131;&#xC219;&#xD55C; &#xC138;&#xD3EC;&#xC5D0;&#xC11C; &#xC5EC;&#xC804;&#xD788; &#xAC10;&#xC9C0;&#xD560; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC720;&#xC120;&#xC758; &#xC21C;&#xD658;&#xC801;&#xC778; &#xAC1C;&#xBC1C; &#xD558;&#xB294; &#xB3D9;&#xC548; Otx1 &#xC720;&#xC804;&#xC790;&#xB294; &#xC138;&#xD3EC; &#xBD84;&#xD654;&#xC5D0;&#xC774; &#xB179;&#xC74C; &#xBC29;&#xC1A1; &#xC694;&#xC778;&#xC758; &#xC5ED;&#xD560;&#xC744; &#xD655;&#xC778; &#xD558;&#xB294; &#xC218; &#xC720;&#xC5D0; overexpressed&#xC785;&#xB2C8;&#xB2E4;. &#xCD5C;&#xADFC; &#xC5F0;&#xAD6C; &#xBCF4;&#xACE0; Otx1 &#xC720;&#xBC29;&#xC554;&#xC5D0; overexpressed&#xC785;&#xB2C8;&#xB2E4;. Otx1&#xC740; embryogenesis, &#xB3D9;&#xC548; &#xD45C;&#xD604; &#xB418; &#xACE0; &#xBC1C;&#xC554;, &#xC885;&#xC591; &#xC57D; &#xC138;&#xD3EC;&#xC758; &#xBD84;&#xD654;&#xC5D0; &#xAC00;&#xB2A5;&#xD55C; &#xAE30;&#xB2A5;&#xC744; &#xC554;&#xC2DC; &#xD558;&#xB294; &#xB3D9;&#xC548; &#xB2E4;&#xC2DC; &#xD45C;&#xC2DC; &#xB429;&#xB2C8;&#xB2E4;.

&#xC720;&#xC120; &#xBC0F; homeobox &#xC720;&#xC804;&#xC790; otx1&#xC785;&#xB2C8;&#xB2E4;.

La gl&aacute;ndula mamaria, el &uacute;nico &oacute;rgano que forman sobre todo en la pubertad, es un modelo ideal para estudiar las funciones de los genes homeobox (HB) en desarrollo y de la tumorig&eacute;nesis. Genes HB forman una gran familia de reguladores del desarrollo que tienen un papel fundamental en el crecimiento celular y diferenciaci&oacute;n. En la gl&aacute;ndula mamaria normal, homeobox genes est&aacute;n involucrados en formaci&oacute;n ductal, ramificaci&oacute;n epitelial y desarrollo lobulo-alveolar mediante la regulaci&oacute;n de la diferenciaci&oacute;n y proliferaci&oacute;n epitelial. Los genes de HB son controlados de manera espacial y temporal en las c&eacute;lulas epiteliales y estromales. Coordinadamente se regulan por las hormonas y la matriz extracelular, lo que sugiere que muchas v&iacute;as de se&ntilde;alizaci&oacute;n est&aacute;n implicadas en las funciones de genes homeobox. Cuando genes homeobox se misexpressed en modelos animales, aparecen diferentes defectos en el desarrollo de la gl&aacute;ndula mamaria. Expresi&oacute;n aberrante de genes homeobox, sobreexpresa o reguladas, se encuentra en carcinomas primarios y en c&aacute;ncer de mama. El gen Otx1 HB es un cl&aacute;sico de regulaci&oacute;n del desarrollo del sistema nervioso durante la embriog&eacute;nesis. Postnatal Otx1 se transcribe en la gl&aacute;ndula pituitaria anterior, donde activa la transcripci&oacute;n de las hormonas hipofisarias y desempe&ntilde;a un papel en la diferenciaci&oacute;n eritroide, hematopoyesis y c&eacute;lulas aumento pluripotentes. Otx1 todav&iacute;a pueden detectarse en las c&eacute;lulas maduras del linaje eritroide y megacaryocytic. Durante el desarrollo c&iacute;clico de la gl&aacute;ndula mamaria, el gen Otx1 se sobreexpresa en lactancia, confirmando un papel de este factor de transcripci&oacute;n en la diferenciaci&oacute;n celular. Estudios recientes informan que Otx1 se sobreexpresa en c&aacute;ncer de mama. Otx1 se expresa durante la embriog&eacute;nesis, y se expresa una vez m&aacute;s durante la carcinog&eacute;nesis, lo que implica su posible funci&oacute;n en la diferenciaci&oacute;n de las c&eacute;lulas neopl&aacute;sicas.

La gl&aacute;ndula mamaria y el gen homeobox Otx1.

Proliferative vitreoretinopathy (PVR) is a severe inflammatory complication of retinal detachment. Pathological epiretinal membranes grow on the retina surface leading to contraction, and surgery fails in 5% to 10% of the cases. We evaluated the expression of VEGF-A, Otx1, Otx2, Otx3, and p53 family members from PVR specimens to correlate their role in inducing or preventing the pathology.

Expression of VEGF-A, Otx homeobox and p53 family genes in proliferative vitreoretinopathy.

Vitreoretinal surgery has advanced in numerous directions during recent years. The removal of the vitreous body is one of the main characteristics of this surgical procedure. Several molecules have been tested in the past to fill the vitreous cavity and to mimic its functions. We here review the currently available vitreous substitutes, focusing on their molecular properties and functions, together with their adverse effects. Afterwards we describe the characteristics of the ideal vitreous substitute. The challenges facing every ophthalmology researcher are to reach a long-term intraocular permanence of vitreous substitute with total inertness of the molecule injected and the control of inflammatory reactions. We report new polymers with gelification characteristics and smart hydrogels representing the future of vitreoretinal surgery. Finally, we describe the current studies on vitreous regeneration and cell cultures to create new intraocular gels with optimal biocompatibility and rheological properties. 

Vitreous substitutes: the present and the future.

Neuronal and inducible nitric oxide synthase (nNOS and iNOS) play a protective and damaging role, respectively, on the intestinal neuromuscular function after ischemia-reperfusion (I/R) injury. To uncover the molecular pathways underlying this dichotomy we investigated their possible correlation with the orthodenticle homeobox proteins OTX1 and OTX2 in the rat small intestine myenteric plexus after in vivo I/R. Homeobox genes are fundamental for the regulation of the gut wall homeostasis both during development and in pathological conditions (inflammation, cancer). I/R injury was induced by temporary clamping the superior mesenteric artery under anesthesia, followed by 24 and 48 h of reperfusion. At 48 h after I/R intestinal transit decreased and was further reduced by -propyl-l-arginine hydrochloride (NPLA), a nNOS-selective inhibitor. By contrast this parameter was restored to control values by 1400W, an iNOS-selective inhibitor. In longitudinal muscle myenteric plexus (LMMP) preparations, iNOS, OTX1, and OTX2 mRNA and protein levels increased at 24 and 48 h after I/R. At both time periods, the number of iNOS- and OTX-immunopositive myenteric neurons increased. nNOS mRNA, protein levels, and neurons were unchanged. In LMMPs, OTX1 and OTX2 mRNA and protein upregulation was reduced by 1400W and NPLA, respectively. In myenteric ganglia, OTX1 and OTX2 staining was superimposed with that of iNOS and nNOS, respectively. Thus in myenteric ganglia iNOS- and nNOS-derived NO may promote OTX1 and OTX2 upregulation, respectively. We hypothesize that the neurodamaging and neuroprotective roles of iNOS and nNOS during I/R injury in the gut may involve corresponding activation of molecular pathways downstream of OTX1 and OTX2. Intestinal ischemia-reperfusion (I/R) injury induces relevant alterations in myenteric neurons leading to dismotility. Nitrergic neurons seem to be selectively involved. In the present study the inference that both neuronal and inducible nitric oxide synthase (nNOS and iNOS) expressing myenteric neurons may undergo important changes sustaining derangements of motor function is reinforced. In addition, we provide data to suggest that NO produced by iNOS and nNOS regulates the expression of the vital transcription factors orthodenticle homeobox protein 1 and 2 during an I/R damage.

Nitric oxide regulates homeoprotein OTX1 and OTX2 expression in the rat myenteric plexus after intestinal ischemia-reperfusion injury.

To evaluate optical coherence tomography (OCT) modifications of outer retinal layers as determinants for functional recovery after surgery for idiopathic macular hole (IMH).

Macular hole surgery: the healing process of outer retinal layers to visual acuity recovery.

OTX Homeobox genes are involved in embryonic morphogenesis and in the development of olfactory epithelium in adult. Mutations occurring in the OTX genes are reported to be associated to tumorigenisis in human. No reports correlate the expression of OTX genes and neoplasms of the nasal cavity. Thus, through immunohistochemical and Real-time PCR analysis we investigated OTX1 and OTX2 expression in the more frequent types of nasal and sinonasal tumours. Variable expression of both genes were found in normal sinonasal mucosa and in tumours. Interestingly, no expression of both OTX genes were detected in sinonasal intestinal-type adenocarcinomas; only OTX1 was found in non-intestinal-type adenocarcinomas and OTX2 was selectively expressed in olfactory neuroblastomas. In conclusion, OTX1 and OTX2 genes might have a role in the pathogenesis of different types of sinonasal neoplasms.

OTX1 and OTX2 as possible molecular markers of sinonasal carcinomas and olfactory neuroblastomas.

Chronic Myeloid Leukemia (CML) is a stem cell cancer that arises when t(9;22) translocation occurs in a hematopoietic stem cells. This event results in the expression of the BCR-ABL1 fusion gene, which codes for a constitutively active tyrosine kinase that is responsible for the transformation of a HSC into a CML stem cell, which then gives rise to a clonal myeloproliferative disease. The introduction of Tyrosine Kinase Inhibitors (TKIs) has revolutionized the management of the disease. However, these drugs do not seem to be able to eradicate the malignancy. Indeed, discontinuation trials (STIM; TWISER; DADI) for those patients who achieved a profound molecular response showed 50% relapsing within 12 months. We performed a comparative analysis on 15 CML patients and one B-ALL patient, between the standard quantitative reverse-transcriptase PCR (qRT-PCR) and our genomic DNA patient-specific quantitative PCR assay (gDNA qPCR). Here we demonstrate that gDNA qPCR is better than standard qRT-PCR in disease monitoring after an average follow-up period of 200 days. Specifically, we statistically demonstrated that DNA negativity is more reliable than RNA negativity in indicating when TKIs therapy can be safely stopped.

Giovanni Porta

Department of Medicine and Surgery

University of Insubria

Identification of OTX1 and OTX2 As Two Possible Molecular Markers for Sinonasal Carcinomas and Olfactory Neuroblastomas

Giovanni Porta

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Medicine and Surgery

University of Insubria

Identification of OTX1 and OTX2 As Two Possible Molecular Markers for Sinonasal Carcinomas and Olfactory Neuroblastomas

Department of Medicine and Surgery