an-optimized-protocol-for-electrophoretic-mobility-shift-assay-using

An Optimized Protocol for Electrophoretic Mobility Shift Assay Using Infrared Fluorescent Dye-labeled Oligonucleotides

Electrophoretic Mobility Shift Assays (EMSA) are an instrumental tool to characterize the interactions between proteins and their target DNA sequences. ...

University of Illinois at Chicago

&#x5728;&#x8FD9;&#x91CC;&#x62A5;&#x544A;&#x7684;&#x4E34;&#x5E8A;&#x524D;&#x7814;&#x7A76;&#xFF0C;VX2 &#x6279;&#x91CF;&#x8D85;&#x58F0;&#x6D88;&#x878D;&#x96C7;&#x7528;&#x5C0F;&#x578B;&#x5316;&#x7684;&#x6CBB;&#x7597;&#x5154;&#x809D;&#x5185;&#x7684;&#x764C;&#x56FE;&#x50CF;-ablate &#x9635;&#x5217;&#x3002;&#x9635;&#x5217;&#x63A2;&#x6D4B;&#x5668;&#x5EFA;&#x9020;&#x4E86;&#x4E0E; 32 &#x4E2D; 2.3 &times; 20 mm(2) &#x5149;&#x5708;&#x3001; &#x5305;&#x88C5;&#x5728;&#x51B7;&#x5374;&#x548C;&#x8026;&#x5408;&#x6C14;&#x7403;&#x4F53;&#x5185;&#x4F7F;&#x7528; 3.1 &#x6BEB;&#x7C73;&#x4E0D;&#x9508;&#x94A2;&#x7BA1;&#x5185;&#x7684;&#x5143;&#x7D20;&#x3002;&#x63A2;&#x6D4B;&#x5668;&#x6D4B;&#x91CF;&#x80FD;&#x529B;&#x7684; 50%&#x5206;&#x6570;&#x5E26;&#x5BBD;&#x7684;&#x8109;&#x51B2;&#x56DE;&#x6CE2;&#x6210;&#x50CF; &#xFF08;&#x4E2D;&#x5FC3;&#x9891;&#x7387; 4.4 MHz) &#x4E0E; &gt; 110 W/cm(2) &#x8868;&#x9762;&#x5F3A;&#x5EA6;&#x8D85;&#x58F0;&#x6CE2;&#x9891;&#x7387; 3.5 &#x548C; 4.8 MHz.B-&#x626B;&#x63CF;&#x6210;&#x50CF;&#x9635;&#x5217;&#x7684;&#x6027;&#x80FD;&#x8861;&#x91CF;&#x53EF;&#x5AB2;&#x7F8E;&#x5927;&#x8BCA;&#x65AD;&#x7684;&#x7EBF;&#x6027;&#x9635;&#x5217;&#xFF0C;&#x5C3D;&#x7BA1;&#x8FD1;&#x573A;&#x56FE;&#x50CF;&#x8D28;&#x91CF;&#x964D;&#x4F4E;&#x4E86;&#x632F;&#x94C3;&#x9879;&#x76EE;&#x516C;&#x5F00;&#x53D1;&#x552E;&#x3002;&#x8FDB;&#x884C;&#x4E86;&#x4E00;&#x7CFB;&#x5217;&#x7684;&#x4F53;&#x5185;&#x6D88;&#x878D;&#x6CBB;&#x7597;&#x7A0B;&#x5E8F;&#xFF0C;&#x4F7F;&#x7528;&#x66DD;&#x5149;&#x5DE5;&#x671F; 20 70.5 s &#x4E0E;&#x539F;&#x4F4D;&#x7A7A;&#x95F4;&#x5E73;&#x5747;&#x3001; &#x65F6;&#x7A7A;&#x5E73;&#x5747;&#x5F3A;&#x5EA6;&#x4E3A; 4.8 MHz &#x53D1;&#x5C04; 22.4 38.5 W/cm(2) &#x91CD;&#x70B9;&#x7684; 32 &#x5143;&#x5B54;&#x5F84;&#x3002;&#x5B8C;&#x6574;&#x7684;&#x80BF;&#x7624;&#x6A2A;&#x622A;&#x9762;&#x7684;&#x6D88;&#x878D;&#x8BC1;&#x5B9E;&#x5728; 12 &#x66DD;&#x5149;&#xFF0C;&#x5E26;&#x56DB;&#x4E2A;&#x98CE;&#x9669;&#x655E;&#x53E3;&#x70E7;&#x8680;&#x8FFD;&#x52A0;&#x4FDD;&#x8BC1;&#x91D1;&#x7684;&#x4E03;&#x4E2A;&#x91CD;&#x8981;&#x67D3;&#x8272; &gt; &#x4EE5;&#x5916;&#x6240;&#x6709;&#x65B9;&#x5411;&#x80BF;&#x7624; 1 &#x6BEB;&#x7C73;&#x3002;&#x5206;&#x6790;&#x8868;&#x660E;&#xFF0C;&#x9608;&#x503C;&#x6D88;&#x878D;&#x6548;&#x679C;&#xFF0C;&#x4E0E;&#x7167;&#x5C04;&#x80BF;&#x7624;&#x5256;&#x9762;&#x4E0E;&#x4EA4;&#x4ED8;&#x7684;&#x5B8C;&#x5168;&#x6D88;&#x878D; &gt; 838 J &#x58F0;&#x6CE2;&#x7684;&#x80FD;&#x91CF;&#x3002;&#x7ED3;&#x679C;&#x663E;&#x793A;&#x53EF;&#x884C;&#x6027;&#x4F53;&#x5185;&#x809D;&#x764C;&#x6D88;&#x878D;&#x4F7F;&#x7528;&#x5C0F;&#x578B;&#x5316;&#x7684;&#x56FE;&#x50CF;-ablate &#x9635;&#x5217;&#x9002;&#x5408;&#x95F4;&#x8D28;&#x90E8;&#x7F72;&#x3002;

&#x6CBB;&#x7597;&#x5154;&#x809D;&#x80BF;&#x7624;&#x5728;&#x4F53;&#x5185;&#x4F7F;&#x7528;&#x5C0F;&#x578B;&#x5316;&#x7684;&#x56FE;&#x50CF; ablate &#x8D85;&#x58F0;&#x9635;&#x5217;&#x3002;

In the preclinical studies reported here, VX2 cancer within rabbit liver has been treated by bulk ultrasound ablation employing miniaturized image-ablate arrays. Array probes were constructed with 32 elements in a 2.3 Ã— 20 mm(2) aperture, packaged within a 3.1 mm stainless steel tube with a cooling and coupling balloon for in vivo use. The probes were measured capable of 50% fractional bandwidth for pulse-echo imaging (center frequency 4.4 MHz) with >110 W/cm(2) surface intensity available at sonication frequencies 3.5 and 4.8 MHz. B-scan imaging performance of the arrays was measured to be comparable to larger diagnostic linear arrays, although nearfield image quality was reduced by ringdown artifacts. A series of in vivo ablation procedures was performed using an unfocused 32-element aperture firing at 4.8 MHz with exposure durations 20-70.5 s and in situ spatial average, temporal average intensities 22.4-38.5 W/cm(2). Ablation of a complete tumor cross-section was confirmed by vital staining in seven of 12 exposures, with four exposures ablating an additional margin >1 mm beyond the tumor in all directions. Analysis suggests a threshold ablation effect, with complete ablation of tumor cross-sections for exposures with delivery of >838 J acoustic energy. The results show feasibility for in vivo liver cancer ablation using miniaturized image-ablate arrays suitable for interstitial deployment.

Treatment of rabbit liver cancer in vivo using miniaturized image-ablate ultrasound arrays.

Dans les &eacute;tudes pr&eacute;cliniques rapport&eacute;es ici, VX2 cancer dans le foie de lapin a &eacute;t&eacute; trait&eacute; en vrac &eacute;chographie ablation employant miniaturis&eacute;s image-ablation tableaux. Tableau des sondes ont &eacute;t&eacute; construits avec 32 &eacute;l&eacute;ments dans une ouverture de mm(2) 2,3 &times; 20, emball&eacute; dans un tube d&#39;acier inoxydable de 3,1 mm avec un refroidissement et un ballon de couplage pour utilisation in vivo. Les sondes ont &eacute;t&eacute; mesur&eacute;s capables de 50 % de bande passante fractionnelle d&#39;imagerie de l&#39;impulsion de l&#39;&eacute;cho (fr&eacute;quence centrale 4,4 MHz) avec &gt; 110 W/cm(2) surface intensit&eacute; disponible &agrave; des fr&eacute;quences de sonication 3,5 &agrave; 4,8 MHz. B-analyse des performances d&#39;imagerie des tableaux a &eacute;t&eacute; mesur&eacute; pour &ecirc;tre comparable aux plus grandes baies lin&eacute;aires diagnostiques, bien que peu &eacute;loign&eacute;e des images de qualit&eacute; a &eacute;t&eacute; r&eacute;duite par les artefacts de sonnerie. Une s&eacute;rie de proc&eacute;dures d&#39;ablation in vivo a &eacute;t&eacute; r&eacute;alis&eacute;e &agrave; l&#39;aide d&#39;une ouverture de 32 &eacute;l&eacute;ments floue tir &agrave; 4,8 MHz avec exposition dur&eacute;e 20-70,5 s et de l&#39;intensit&eacute; spatiale moyenne, temporelle moyenne in situe 22,4-38,5 W/cm(2). Ablation d&#39;un &eacute;ventail complet de tumeur a &eacute;t&eacute; confirm&eacute;e par une coloration vitale dans sept des 12 expositions, avec quatre expositions ablation une marge additionnelle &gt; 1 mm au-del&agrave; de la tumeur dans toutes les directions. Analyse sugg&egrave;re un effet de seuil ablation, avec ablation compl&egrave;te des sections transversales de tumeur pour les expositions avec livraison de &gt; 838 &eacute;nergie acoustique de J. Les r&eacute;sultats montrent la faisabilit&eacute; pour un cancer du foie in vivo ablation &agrave; l&#39;aide de miniaturis&eacute;s image-ablation tableaux adapt&eacute;s pour un d&eacute;ploiement interstitiel.

Traitement du cancer de foie de lapin in vivo utilisant miniaturis&eacute;s image-ablation baies de l&#39;&eacute;chographie.

In der pr&auml;klinischen Studien hier berichtet VX2 Krebs innerhalb Kaninchen Leber behandelt worden ist, durch den Massenimport Ultraschall Ablation Einsatz miniaturisierte Bild-entfernen Arrays. Array-Sensoren wurden mit 32 Elemente in einem 2,3 &times; 20 mm(2) Blende, verpackt in ein 3,1 mm Edelstahlrohr mit K&uuml;hlung und Kupplung Ballon f&uuml;r in-vivo Gebrauch konstruiert. Die Sonden wurden von 50 % gebrochene Bandbreite f&uuml;r Impuls-Echo-Bildgebung gemessen (Mittenfrequenz 4,4 MHz) mit &gt; 110 W/cm(2) Oberfl&auml;che Intensit&auml;t unter Anwendung von Ultraschall-Frequenzen 3,5 und 4,8 MHz. B-Scan Abbildungsleistung der Arrays gemessen wurde, um gr&ouml;&szlig;ere diagnostische lineare Arrays, vergleichbar zu sein, obwohl Opische Bildqualit&auml;t wurde durch Ringdown Artefakte reduziert. Eine Reihe von in-vivo-Ablation Verfahren wurde mithilfe einer unscharf 32-Element-Blende feuern bei 4,8 MHz mit Exposition Dauer 20-70,5 s und in-situ r&auml;umliche durchschnittliche, zeitliche durchschnittliche Intensit&auml;ten 22,4-38,5 W/cm(2) durchgef&uuml;hrt. Ablation eine komplette Tumor-Querschnitt von lebenswichtigen F&auml;rbung in sieben der 12 Forderungen, mit vier Forderungen Abtragen eine zus&auml;tzliche Marge best&auml;tigt &gt; 1 mm &uuml;ber den Tumor in alle Richtungen. Analyse zufolge ein Schwellwert Abtragen, mit kompletten Ablation der Tumor Querschnitte f&uuml;r Forderungen mit der Lieferung von &gt; 838 J akustische Energie. Die Ergebnisse zeigen die Machbarkeit f&uuml;r in-vivo Leberkrebs Ablation mit miniaturisierte Bild-entfernen Arrays f&uuml;r interstitielle Deployment.

Behandlung von Kaninchen Leber Krebs in Vivo mit miniaturisierte Bild-Ultraschall-Arrays entfernen.

Negli studi preclinici qui riportati, VX2 cancro all&#39;interno del fegato di coniglio &egrave; stato trattato impiegando bulk ecografia ablazione miniaturizzati immagine-ablazione matrici. Sonde array sono state costruite con 32 elementi in un&#39;apertura mm(2) 2,3 &times; 20, confezionato all&#39;interno di un tubo di acciaio inox 3,1 mm con un palloncino di accoppiamento per l&#39;uso in vivo e di raffreddamento. Le sonde sono state misurate in grado di 50% di banda frazionale per imaging pulse-echo (centro frequenza MHz 4,4) con &amp;gt; 110 W/cm(2) superficie intensit&agrave; disponibile a frequenze di sonicazione 3,5 e 4,8 MHz. B-scan imaging prestazioni delle matrici &egrave; stata misurata per essere paragonabile ai pi&ugrave; grandi matrici lineari diagnostici, sebbene nearfield la qualit&agrave; dell&#39;immagine &egrave; stata ridotta da ringdown artefatti. Una serie di procedure di ablazione in vivo &egrave; stata eseguita utilizzando un diaframma 32-elemento unfocused sparando a 4,8 MHz con esposizione durate 20-70.5 s e in situ spaziale medio, temporale media intensit&agrave; 22,4-38,5 W/cm(2). Ablazione di una sezione trasversale completa del tumore &egrave; stata confermata dalla colorazione vitale in sette delle 12 esposizioni, con quattro esposizioni un margine supplementare di ablazione &amp;gt; 1 mm oltre il tumore in tutte le direzioni. L&#39;analisi suggerisce un effetto di ablazione di soglia, con ablazione completa di sezioni trasversali del tumore per le esposizioni con la consegna di &amp;gt; 838 energia acustica J. I risultati mostrano la fattibilit&agrave; per il cancro del fegato in vivo ablazione utilizzando miniaturizzati immagine-ablazione matrici adatti per distribuzione interstiziale.

Trattamento del cancro del fegato di coniglio in vivo utilizzando miniaturizzati immagine-ablazione matrici ad ultrasuoni.

&#x30A2;&#x30EC;&#x30A4;&#x753B;&#x50CF;&#x713C;&#x707C; VX2 &#x764C;&#x5BB6;&#x514E;&#x809D;&#x5185;&#x30D0;&#x30EB;&#x30AF;&#x8D85;&#x97F3;&#x6CE2;&#x30A2;&#x30D6;&#x30EC;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x5C0F;&#x578B;&#x63A1;&#x7528;&#x306B;&#x3088;&#x308A;&#x6CBB;&#x7642;&#x3055;&#x308C;&#x3066;&#x3044;&#x308B;, &#x3053;&#x3053;&#x306B;&#x5831;&#x544A;&#x524D;&#x81E8;&#x5E8A;&#x7814;&#x7A76;&#x3002;&#x30A2;&#x30EC;&#x30A4; &#x30D7;&#x30ED;&#x30FC;&#x30D6;&#x306F;&#x3001;2.3 &times; 20 mm(2) &#x53E3;&#x5F84;&#x306E; 3.1 mm &#x30B9;&#x30C6;&#x30F3;&#x30EC;&#x30B9;&#x92FC;&#x7BA1;&#x51B7;&#x5374;&#x3068; in vivo &#x3067;&#x306E;&#x4F7F;&#x7528;&#x306E;&#x305F;&#x3081;&#x306E;&#x30AB;&#x30C3;&#x30D7;&#x30EA;&#x30F3;&#x30B0; &#x30D0;&#x30EB;&#x30FC;&#x30F3;&#x5185;&#x306E;&#x30D1;&#x30C3;&#x30B1;&#x30FC;&#x30B8;&#x306E; 32 &#x8981;&#x7D20;&#x3067;&#x5EFA;&#x8A2D;&#x3055;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;&#x30D7;&#x30ED;&#x30FC;&#x30D6;&#x306F; 50% &#x306E;&#x6BD4;&#x5E2F;&#x57DF;&#x5E45;&#x30D1;&#x30EB;&#x30B9;-&#x30A8;&#x30B3;&#x30FC; &#x30A4;&#x30E1;&#x30FC;&#x30B8;&#x30F3;&#x30B0;&#x306E;&#x305F;&#x3081;&#x306E;&#x53EF;&#x80FD;&#x306A;&#x6E2C;&#x5B9A;&#x3057;&#x305F; (&#x4E2D;&#x5FC3;&#x5468;&#x6CE2;&#x6570; 4.4 MHz) &#x3068;&#x76EE; 110 W/cm(2) &#x8868;&#x9762;&#x30A4;&#x30F3;&#x30C6;&#x30F3;&#x30B7;&#x30C6;&#x30A3; 3.5 &#x3068; 4.8 MHz&#x3002; B &#x30A4;&#x30E1;&#x30FC;&#x30B8;&#x30F3;&#x30B0; &#x30A2;&#x30EC;&#x30A4;&#x306E;&#x30D1;&#x30D5;&#x30A9;&#x30FC;&#x30DE;&#x30F3;&#x30B9;&#x306F;&#x3001;&#x8FD1;&#x8DDD;&#x96E2;&#x753B;&#x50CF;&#x304C;&#x54C1;&#x8CEA;&#x8A3A;&#x65AD;&#x306E;&#x5927;&#x898F;&#x6A21;&#x306E;&#x7DDA;&#x5F62;&#x30A2;&#x30EC;&#x30A4;&#x306B;&#x5339;&#x6575;&#x3059;&#x308B;&#x3068;&#x6E2C;&#x5B9A;&#x3055;&#x308C;&#x305F;&#x30B9;&#x30AD;&#x30E3;&#x30F3; &#x30EA;&#x30F3;&#x30B0;&#x30C0;&#x30A6;&#x30F3; &#x30A2;&#x30FC;&#x30C6;&#x30A3;&#x30D5;&#x30A1;&#x30AF;&#x30C8;&#x306B;&#x3088;&#x3063;&#x3066;&#x524A;&#x6E1B;&#x3055;&#x308C;&#x305F;&#x8D85;&#x97F3;&#x6CE2;&#x5468;&#x6CE2;&#x6570;&#x3067;&#x5229;&#x7528;&#x3067;&#x304D;&#x307E;&#x3059;&#x3002;&#x4E00;&#x9023;&#x306E; in vivo &#x3067;&#x306E;&#x30AB;&#x30C6;&#x30FC;&#x30C6;&#x30EB;&#x30A2;&#x30D6;&#x30EC;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x306E;&#x624B;&#x9806;&#x306F;&#x3001;4.8 MHz &#x3067;&#x66B4;&#x9732;&#x671F;&#x9593; 20-70.5 s &#x53CA;&#x3073; in situ &#x7A7A;&#x9593;&#x5E73;&#x5747;&#x3001;&#x6642;&#x9593;&#x7684;&#x5E73;&#x5747;&#x5F37;&#x5EA6;&#x3068;&#x767A;&#x706B; 22.4 38.5 W/cm(2) &#x3084;&#x308A;&#x5834;&#x306E; 32 &#x8981;&#x7D20;&#x7D5E;&#x308A;&#x3092;&#x7528;&#x3044;&#x3066;&#x884C;&#x3063;&#x305F;&#x3002;&#x30A2;&#x30D6;&#x30EC;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x816B;&#x760D;&#x5B8C;&#x5168;&#x65AD;&#x9762;&#x306E; 12 &#x306E;&#x30A8;&#x30AF;&#x30B9;&#x30DD;&#x30FC; &#x30B8;&#x30E3;&#x30FC;&#x3001;&#x8FFD;&#x52A0;&#x30DE;&#x30FC;&#x30B8;&#x30F3;&#x3092;&#x30A2;&#x30D6;&#x30EC;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x3060; 4 &#x3064;&#x306E;&#x30A8;&#x30AF;&#x30B9;&#x30DD;&#x30FC; &#x30B8;&#x30E3;&#x30FC;&#x306E; 7 &#x3064;&#x306E;&#x751F;&#x4F53;&#x67D3;&#x8272;&#x306B;&#x3088;&#x3063;&#x3066;&#x78BA;&#x8A8D;&#x3055;&#x308C;&#x305F;&#x76EE; 1 &#x30DF;&#x30EA;&#x30E1;&#x30FC;&#x30C8;&#x30EB;&#x3092;&#x8D85;&#x3048;&#x3066;&#x3059;&#x3079;&#x3066;&#x306E;&#x65B9;&#x5411;&#x306B;&#x816B;&#x760D;&#x3002;&#x5206;&#x6790;&#x306F;&#x3001;&#x816B;&#x760D;&#x306E;&#x65AD;&#x9762;&#x306E;&#x30A8;&#x30AF;&#x30B9;&#x30DD;&#x30FC; &#x30B8;&#x30E3;&#x30FC;&#x306E;&#x305F;&#x3081;&#x306E;&#x5B8C;&#x5168;&#x306A;&#x30A2;&#x30D6;&#x30EC;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x306E;&#x914D;&#x4FE1;&#x3068;&#x306E;&#x3057;&#x304D;&#x3044;&#x5024;&#x30A2;&#x30D6;&#x30EC;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x52B9;&#x679C;&#x3092;&#x793A;&#x5506;&#x76EE; 838 J &#x97F3;&#x97FF;&#x30A8;&#x30CD;&#x30EB;&#x30AE;&#x30FC;&#x3002;&#x7D50;&#x679C;&#x5B9F;&#x73FE;&#x53EF;&#x80FD;&#x6027;&#x3092;&#x793A;&#x3059; in vivo &#x3067;&#x306E;&#x809D;&#x81D3;&#x304C;&#x3093;&#x306E;&#x305F;&#x3081;&#x5C0F;&#x578B;&#x30A2;&#x30D6;&#x30EC;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x3092;&#x7528;&#x3044;&#x305F;&#x753B;&#x50CF;-&#x713C;&#x707C;&#x30A2;&#x30EC;&#x30A4;&#x9593;&#x8CEA;&#x6027;&#x306E;&#x5C55;&#x958B;&#x306E;&#x305F;&#x3081;&#x306B;&#x9069;&#x3057;&#x305F;&#x3002;

Vivo &#x5C0F;&#x578B;&#x5316;&#x3092;&#x7528;&#x3044;&#x305F;&#x30A6;&#x30B5;&#x30AE;&#x809D;&#x81D3;&#x304C;&#x3093;&#x306E;&#x6CBB;&#x7642;&#x306E;&#x30A4;&#x30E1;&#x30FC;&#x30B8;-&#x713C;&#x707C;&#x8D85;&#x97F3;&#x6CE2;&#x30A2;&#x30EC;&#x30A4;&#x3002;

&#xC5EC;&#xAE30;&#xC5D0; &#xBCF4;&#xACE0; &#xD558;&#xB294; &#xC804; &#xC784;&#xC0C1; &#xC5F0;&#xAD6C;&#xC5D0;&#xC11C; VX2 &#xB300;&#xB7C9; &#xCD08;&#xC74C;&#xD30C; &#xC808;&#xC81C; &#xACE0;&#xC6A9; &#xCD95;&#xC18C; &#xC2DC;&#xD0A8; &#xD1A0;&#xB07C; &#xAC04; &#xB0B4; &#xC554; &#xCC98;&#xB9AC; &#xC774;&#xBBF8;&#xC9C0;-&#xC808;&#xC81C; &#xBC30;&#xC5F4;. &#xBC30;&#xC5F4; &#xD504;&#xB85C;&#xBE0C;&#xB294; &#xB0C9;&#xAC01; &#xBC0F; &#xC0DD;&#xCCB4; &#xC870;&#xAC74; &#xC0AC;&#xC6A9;&#xC5D0; &#xB300; &#xD55C; &#xCEE4;&#xD50C;&#xB9C1; &#xD48D;&#xC120; 3.1 m m &#xC2A4;&#xD14C;&#xC778;&#xB808;&#xC2A4; &#xC2A4;&#xD2F8; &#xD29C;&#xBE0C; &#xB0B4;&#xC5D0; &#xD328;&#xD0A4;&#xC9C0;&#xD654; 2.3 &times; 20 mm(2) &#xC870;&#xB9AC;&#xAC1C;&#xC758; &#xC694;&#xC18C;&#xC5D0; 32 &#xAC74;&#xC124; &#xB418;&#xC5C8;&#xB2E4;. &#xD504;&#xB85C;&#xBE0C; &#xD384;&#xC2A4;-&#xC5D0;&#xCF54; &#xC774;&#xBBF8;&#xC9D5;&#xC5D0; &#xB300; &#xD55C; 50% &#xC18C;&#xC218; &#xB300;&#xC5ED;&#xD3ED;&#xC758; &#xCE21;&#xC815; (&#xC8FC;&#xD30C;&#xC218; 4.4 m h z)&#xC640; &amp;gt; 110 W/cm(2) &#xD45C;&#xBA74; &#xAC15;&#xB3C4; 3.5 &#xBC0F; 4.8 m h z. B &#xC2A4;&#xCE94; nearfield &#xC774;&#xBBF8;&#xC9C0; &#xD488;&#xC9C8; &#xBE44;&#xB85D; &#xD070; &#xC9C4;&#xB2E8; &#xC120;&#xD615; &#xBC30;&#xC5F4;&#xACFC; &#xBE44;&#xAD50; &#xB420; &#xBC30;&#xC5F4; &#xC774;&#xBBF8;&#xC9D5; &#xC131;&#xB2A5; &#xCE21;&#xC815; ringdown &#xC544;&#xD2F0;&#xD329;&#xD2B8;&#xC5D0; &#xC758;&#xD574; &#xAC10;&#xC18C; &#xD588;&#xB2E4; sonication &#xC8FC;&#xD30C;&#xC218;&#xC5D0;&#xC11C; &#xC0AC;&#xC6A9;&#xD560; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC77C;&#xB828;&#xC758; &#xC0DD;&#xCCB4; &#xC870;&#xAC74; &#xC81C;&#xAC70; &#xD504;&#xB85C;&#xC2DC;&#xC800;&#xAC00; 22.4 38.5 W/cm(2) &#xD604;&#xC7A5;&#xC5D0;&#xC11C; &#xACF5;&#xAC04; &#xD3C9;&#xADE0;, &#xC2DC;&#xAC04;&#xC801;&#xC778; &#xD3C9;&#xADE0; &#xB18D;&#xB3C4;&#xC640; &#xB178;&#xCD9C; &#xAE30;&#xAC04; 20-70.5 s 4.8 m h Z&#xC5D0;&#xC11C; &#xBC1C;&#xC0AC; &#xC0B0;&#xB9CC;&#xB41C; 32 &#xC694;&#xC18C; &#xC870;&#xB9AC;&#xAC1C;&#xB97C; &#xC0AC;&#xC6A9; &#xD558; &#xC5EC; &#xC218;&#xD589; &#xB418;&#xC5C8;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC644;&#xC804; &#xD55C; &#xC885;&#xC591; &#xD6A1;&#xB2E8;&#xBA74;&#xC758; &#xC808;&#xC81C; 7 4 &#xB178;&#xCD9C; ablating &#xCD94;&#xAC00; &#xB9C8;&#xC9C4; 12 &#xB178;&#xCD9C; &#xC911;&#xC694; &#xD55C; &#xC5BC;&#xB8E9;&#xC5D0; &#xC758;&#xD574; &#xD655;&#xC778; &#xB410;&#xB2E4; &amp;gt; &#xBAA8;&#xB4E0; &#xBC29;&#xD5A5;&#xC5D0;&#xC11C; &#xC885;&#xC591;&#xC744; &#xB118;&#xC5B4; 1 m m. &#xBD84;&#xC11D; &#xC81C;&#xC548; &#xBC30;&#xB2EC;&#xC758; &#xB178;&#xCD9C;&#xC5D0; &#xB300; &#xD55C; &#xD6A1;&#xB2E8;&#xBA74; &#xC885;&#xC591;&#xC758; &#xC644;&#xC804; &#xC808;&#xC81C;&#xC640; &#xC784;&#xACC4;&#xAC12; &#xC808;&#xC81C; &#xD6A8;&#xACFC; &amp;gt; 838 J &#xC5B4;&#xCFE0;&#xC2A4;&#xD2F1; &#xC5D0;&#xB108;&#xC9C0;. &#xACB0;&#xACFC; &#xD0C0;&#xB2F9;&#xC131;&#xC744; &#xBCF4;&#xC5EC; vivo&#xC5D0;&#xC11C; &#xAC04; &#xC554;&#xC5D0; &#xB300; &#xD55C; &#xC808;&#xC81C;&#xB97C; &#xC0AC;&#xC6A9; &#xD558; &#xC5EC; &#xCD95;&#xC18C; &#xC2DC;&#xD0A8; &#xC774;&#xBBF8;&#xC9C0;-&#xC808;&#xC81C; &#xBC30;&#xC5F4; &#xC911;&#xAC04; &#xBC30;&#xD3EC;&#xC5D0; &#xC801;&#xD569; &#xD569;&#xB2C8;&#xB2E4;.

&#xBE44;&#xBCF4;&#xB97C; &#xC0AC;&#xC6A9; &#xD558; &#xC5EC; &#xCD95;&#xC18C; &#xC2DC;&#xD0A8; &#xD1A0;&#xB07C; &#xAC04; &#xC554; &#xCE58;&#xB8CC;&#xC758; &#xC774;&#xBBF8;&#xC9C0;-&#xC808;&#xC81C; &#xCD08;&#xC74C;&#xD30C; &#xBC30;&#xC5F4;.

En los estudios precl&iacute;nicos divulgados aqu&iacute;, VX2 c&aacute;ncer en el h&iacute;gado del conejo ha sido tratado por ablaci&oacute;n ultrasonido a granel que emplean miniaturizados imagen-ablar matrices. Sondas de matriz se construyeron con 32 elementos en una abertura de mm(2) 2.3 &times; 20, empaquetado dentro de un tubo de acero de 3.1 mm con un enfriamiento y globo de acoplamiento para uso in vivo. Las sondas se midieron la capacidad de ancho de banda fraccionaria de 50% para la proyecci&oacute;n de imagen de pulso-eco (frecuencia de centro 4.4 MHz) con &gt; 110 intensidad de w/cm(2) superficial disponible en frecuencias de sonicaci&oacute;n 3.5 y 4.8 MHz. B-scan de rendimiento de los arreglos de discos se midi&oacute; para ser comparables a arreglos lineales diagn&oacute;sticos m&aacute;s grandes, aunque la calidad de la imagen de campo cercano se redujo en ringdown artefactos. Una serie de procedimientos de ablaci&oacute;n en vivo se realiz&oacute; con una abertura de 32 elementos disparar a 4.8 MHz con exposici&oacute;n duraci&oacute;n 20-70,5 s e intensidades promedio promedio, temporales espaciales in situ 22,4-38.5 W/cm(2). Ablaci&oacute;n de una secci&oacute;n completa del tumor fue confirmada por la coloraci&oacute;n vital en siete de 12 exposiciones, con cuatro exposiciones ablaci&oacute;n un margen adicional &gt; 1 mm m&aacute;s all&aacute; del tumor en todas las direcciones. An&aacute;lisis sugiere un efecto de la ablaci&oacute;n del umbral, con la ablaci&oacute;n completa de cortes transversales del tumor para exposiciones con entrega de &gt; 838 energ&iacute;a ac&uacute;stica de J. Los resultados demuestran la viabilidad para el c&aacute;ncer de h&iacute;gado en vivo ablaci&oacute;n usando miniaturizados imagen-ablar matrices adecuadas para su implementaci&oacute;n intersticial.

Tratamiento del c&aacute;ncer de h&iacute;gado de conejo en vivo usando miniaturizados de imagen-ablar matrices de ultrasonido.

Research into conditions that improve axon regeneration has the potential to open a new door for treatment of brain injury caused by stroke and neurodegenerative diseases of aging, such as Alzheimer, by harnessing intrinsic neuronal ability to reorganize itself. Elucidating the molecular mechanisms of axon regeneration should shed light on how this process becomes restricted in the postnatal stage and in CNS and therefore could provide therapeutic targets for developing strategy to improve axon regeneration in adult CNS. In this review, we first discuss the general view about nerve regeneration and the advantages of using C. elegans as a model system to study axon regeneration. We then compare the conserved regeneration patterns and molecular mechanisms between C. elegans and vertebrates. Lastly, we discuss the power of femtosecond laser technology and its application in axon regeneration research.

C. elegans as a genetic model to identify novel cellular and molecular mechanisms underlying nervous system regeneration.

Left-right asymmetry in anatomical structures and functions of the nervous system is present throughout the animal kingdom. For example, language centers are localized in the left side of the human brain, while spatial recognition functions are found in the right hemisphere in the majority of the population. Disruption of asymmetry in the nervous system is correlated with neurological disorders. Although anatomical and functional asymmetries are observed in mammalian nervous systems, it has been a challenge to identify the molecular basis of these asymmetries. C. elegans has emerged as a prime model organism to investigate molecular asymmetries in the nervous system, as it has been shown to display functional asymmetries clearly correlated to asymmetric distribution and regulation of biologically relevant molecules. Small non-coding RNAs have been recently implicated in various aspects of neural development. Here, we review cases in which microRNAs are crucial for establishing left-right asymmetries in the C. elegans nervous system. These studies may provide insight into how molecular and functional asymmetries are established in the human brain.

microRNA function in left-right neuronal asymmetry: perspectives from C. elegans.

Previous work indicated that ultrasound echo decorrelation imaging can track and quantify changes in echo signals to predict thermal damage during in vitro radiofrequency ablation (RFA). In the in vivo studies reported here, the feasibility of using echo decorrelation imaging as a treatment monitoring tool was assessed. RFA was performed on normal swine liver (N = 5), and ultrasound ablation using image-ablate arrays was performed on rabbit liver implanted with VX2 tumors (N = 2). Echo decorrelation and integrated backscatter were computed from Hilbert transformed pulse-echo data acquired during RFA and ultrasound ablation treatments. Receiver operating characteristic (ROC) curves were employed to assess the ability of echo decorrelation imaging and integrated backscatter to predict ablation. Area under the ROC curves (AUROC) was determined for RFA and ultrasound ablation using echo decorrelation imaging. Ablation was predicted more accurately using echo decorrelation imaging (AUROC = 0.832 and 0.776 for RFA and ultrasound ablation, respectively) than using integrated backscatter (AUROC = 0.734 and 0.494).

In vivo thermal ablation monitoring using ultrasound echo decorrelation imaging.

The assembly of functional neural circuits is critical for complex thoughts, behavior and general brain function. Precise construction of neural circuits requires orderly transition of sequential events from axon outgrowth, pathfinding, branching, to synaptogenesis. Each of these steps is required to be tightly regulated in order to achieve meticulous formation of neuronal connections. MicroRNAs (miRNAs), which silence gene expression post-transcriptionally via either inhibition of translation or destabilization of messenger RNAs, have emerged as key regulators of neuronal connectivity. The expression of miRNAs in neurons is often temporally and spatially regulated, providing critical timing and local mechanisms that prime neuronal growth cones for dynamic responses to extrinsic cues. Here we summarize recent findings of miRNA regulation of neuronal connectivity in a variety of experimental platforms.

The role of microRNAs in regulating neuronal connectivity.

Asymmetries in the nervous system have been observed throughout the animal kingdom. Deviations of brain asymmetries are associated with a variety of neurodevelopmental disorders; however, there has been limited progress in determining how normal asymmetry is established in vertebrates. In the Caenorhabditis elegans chemosensory system, two pairs of morphologically symmetrical neurons exhibit molecular and functional asymmetries. This review focuses on the development of antisymmetry of the pair of amphid wing "C" (AWC) olfactory neurons, from transcriptional regulation of general cell identity, establishment of asymmetry through neural network formation and calcium signaling, to the maintenance of asymmetry throughout the life of the animal. Many of the factors that are involved in AWC development have homologs in vertebrates, which may potentially function in the development of vertebrate brain asymmetry.

Asymmetric neural development in the Caenorhabditis elegans olfactory system.

A molecular link between distinct neuronal asymmetries.

Diversification of neuron classes is essential for functions of the olfactory system, but the underlying mechanisms that generate individual olfactory neuron types are only beginning to be understood. Here we describe a role of the highly conserved HMG-box transcription factor SOX-2 in postmitotic specification and alternative differentiation of the Caenorhabditis elegans AWC and AWB olfactory neurons. We show that SOX-2 partners with different transcription factors to diversify postmitotic olfactory cell types. SOX-2 functions cooperatively with the OTX/OTD transcription factor CEH-36 to specify an AWC "ground state," and functions with the LIM homeodomain factor LIM-4 to suppress this ground state and drive an AWB identity instead. Our findings provide novel insights into combinatorial codes that drive terminal differentiation programs in the nervous system and reveal a biological function of the deeply conserved Sox2 protein that goes beyond its well-known role in stem cell biology.

Postmitotic diversification of olfactory neuron types is mediated by differential activities of the HMG-box transcription factor SOX-2.

Sox2 goes beyond stem cell biology.

The C. elegans AWC olfactory neuron pair communicates to specify asymmetric subtypes AWCOFF and AWCON in a stochastic manner. Intercellular communication between AWC and other neurons in a transient NSY-5 gap junction network antagonizes voltage-activated calcium channels, UNC-2 (CaV2) and EGL-19 (CaV1), in the AWCON cell, but how calcium signaling is downregulated by NSY-5 is only partly understood. Here, we show that voltage- and calcium-activated SLO BK potassium channels mediate gap junction signaling to inhibit calcium pathways for asymmetric AWC differentiation. Activation of vertebrate SLO-1 channels causes transient membrane hyperpolarization, which makes it an important negative feedback system for calcium entry through voltage-activated calcium channels. Consistent with the physiological roles of SLO-1, our genetic results suggest that slo-1 BK channels act downstream of NSY-5 gap junctions to inhibit calcium channel-mediated signaling in the specification of AWCON. We also show for the first time that slo-2 BK channels are important for AWC asymmetry and act redundantly with slo-1 to inhibit calcium signaling. In addition, nsy-5-dependent asymmetric expression of slo-1 and slo-2 in the AWCON neuron is necessary and sufficient for AWC asymmetry. SLO-1 and SLO-2 localize close to UNC-2 and EGL-19 in AWC, suggesting a role of possible functional coupling between SLO BK channels and voltage-activated calcium channels in AWC asymmetry. Furthermore, slo-1 and slo-2 regulate the localization of synaptic markers, UNC-2 and RAB-3, in AWC neurons to control AWC asymmetry. We also identify the requirement of bkip-1, which encodes a previously identified auxiliary subunit of SLO-1, for slo-1 and slo-2 function in AWC asymmetry. Together, these results provide an unprecedented molecular link between gap junctions and calcium pathways for terminal differentiation of olfactory neurons.

SLO BK Potassium Channels Couple Gap Junctions to Inhibition of Calcium Signaling in Olfactory Neuron Diversification.

Left-right asymmetry in the nervous system is observed across species. Defects in left-right cerebral asymmetry are linked to several neurological diseases, but the molecular mechanisms underlying brain asymmetry in vertebrates are still not very well understood. The Caenorhabditis elegans left and right amphid wing 'C' (AWC) olfactory neurons communicate through intercellular calcium signalling in a transient embryonic gap junction neural network to specify two asymmetric subtypes, AWC(OFF) (default) and AWC(ON) (induced), in a stochastic manner. Here, we highlight the molecular mechanisms that establish and maintain stochastic AWC asymmetry. As the components of the AWC asymmetry pathway are highly conserved, insights from the model organism C. elegans may provide a window onto how brain asymmetry develops in humans.This article is part of the themed issue 'Provocative questions in left-right asymmetry'.

Amel Alqadah

Department of Biological Sciences

University of Illinois at Chicago

An Optimized Protocol for Electrophoretic Mobility Shift Assay Using Infrared Fluorescent Dye-labeled Oligonucleotides