Operating Procedures of the Electrochemotherapy for Treatment of Tumor in Dogs and Cats

Electrochemotherapy (ECT) is a local approach which is used for treating solid tumors of different histologies. Its mechanism is based on cell membrane ...

University of Ljubljana

Clinical and laboratory features of the first detected cases of A. phagocytophila infections in dogs from Slovenia.

Canine babesiosis, caused by intraerythrocytic Babesia spp., is a tick-borne disease of worldwide importance. No information on canine babesiosis has been documented in Slovenia. Therefore, 238 dogs admitted to the Small animal clinic in Ljubljana from the years 2000 to 2002 were tested for the presence of babesial parasites in the blood. Based on clinical, microscopic and molecular investigations, 14 dogs (5.9%) were determined as being infected with babesiae. Clinical signs relating to acute haemolysis, fever, anorexia, depression and haematological abnormalities such as anaemia and thrombocytopenia were noticed in most of the 14 infected dogs. The morphology of the parasites was indicative of Babesia canis infection. Two subspecies were detected, namely B. canis canis (11 dogs, 4.6%) and B. canis vogeli (3 dogs, 1.3%) using PCR and subsequent sequence analysis of portions of nns rRNA gene. In addition, based on nucleotide sequence analysis, the 11 isolates of B. c. canis could be subdivided into three groups, whereas the three B. c. vogeli isolates were genetically identical. The results of this study demonstrate the presence of canine babesiosis due to B. c. canis and B. c. vogeli in Slovenia.

Canine babesiosis in Slovenia: molecular evidence of Babesia canis canis and Babesia canis vogeli.

Effektive Behandlung von perianalen Tumoren bei Hunden mit Elektrochemotherapie.

Electrochemotherapy &egrave; una terapia antitumorale che utilizza corrente continua intensa recapitati localmente, breve impulso elettrico al nodulo tumorale pi&ugrave; chemioterapia. Lo scopo del presente studio era di valutare il trattamento electrochemotherapy dei tumori perianali di diverse dimensioni in cani.

Efficacia trattamento di tumori perianali nei cani con electrochemotherapy.

Electrochemotherapy&#xB294; antitumor &#xCE58;&#xB8CC; &#xC885;&#xC591; &#xACB0; &#xC808; &#xD50C;&#xB7EC;&#xC2A4; &#xD654;&#xD559; &#xC694;&#xBC95;&#xC5D0; &#xB85C;&#xCEEC;&#xB85C; &#xBC30;&#xB2EC;, &#xC9E7;&#xC740; &#xAC15;&#xB82C;&#xD55C; &#xC9C1;&#xB958; &#xC804;&#xAE30; &#xD384;&#xC2A4;&#xB97C; &#xC774;&#xC6A9; &#xD558;&#xB294;. &#xD604;&#xC7AC;&#xC758; &#xC5F0;&#xAD6C;&#xC758; &#xBAA9;&#xD45C;&#xB294; &#xAC1C;&#xC5D0; &#xC788;&#xB294; &#xB2E4;&#xC591; &#xD55C; &#xD06C;&#xAE30;&#xC758; &#xD56D;&#xBB38; &#xC8FC;&#xC704; &#xC885;&#xC591;&#xC758; electrochemotherapy &#xCE58;&#xB8CC;&#xB97C; &#xD3C9;&#xAC00; &#xD588;&#xB2E4;.

Electrochemotherapy &#xAC00;&#xC9C4; &#xAC1C;&#xC5D0;&#xC11C; &#xD56D;&#xBB38; &#xC8FC;&#xC704; &#xC885;&#xC591;&#xC758; &#xD6A8;&#xACFC;&#xC801;&#xC778; &#xCE58;&#xB8CC;.

Electrochemotherapy es una terapia antitumoral que utiliza el impulso el&eacute;ctrico entregado localmente, corta corriente intensa al n&oacute;dulo tumoral m&aacute;s quimioterapia. El objetivo del presente estudio fue evaluar el tratamiento de la electrochemotherapy de tumores perianales de diferentes tama&ntilde;os en perros.

Tratamiento efectivo de tumores perianales en los perros con electrochemotherapy.

&#x96FB;&#x6C17;&#x5316;&#x5B66;&#x7642;&#x6CD5;&#x306B;&#x3088;&#x308B;&#x816B;&#x760D;&#x7D50;&#x7BC0;&#x306B;&#x52A0;&#x3048;&#x3066;&#x5316;&#x5B66;&#x7642;&#x6CD5;&#x306B;&#x30ED;&#x30FC;&#x30AB;&#x30EB;&#x306B;&#x914D;&#x4FE1;&#x3055;&#x308C;&#x308B;&#x3001;&#x77ED;&#x3044;&#x5F37;&#x70C8;&#x306A;&#x76F4;&#x6D41;&#x96FB;&#x6C17;&#x30D1;&#x30EB;&#x30B9;&#x3092;&#x5229;&#x7528;&#x3057;&#x3066;&#x3001;&#x6297;&#x816B;&#x760D;&#x7642;&#x6CD5;&#x3067;&#x3059;&#x3002;&#x672C;&#x7814;&#x7A76;&#x306E;&#x76EE;&#x7684;&#x306F;&#x3001;&#x7570;&#x306A;&#x308B;&#x30B5;&#x30A4;&#x30BA;&#x306E;&#x72AC;&#x306E;&#x809B;&#x9580;&#x5468;&#x56F2;&#x816B;&#x760D;&#x306E;&#x96FB;&#x6C17;&#x5316;&#x5B66;&#x7642;&#x6CD5;&#x306B;&#x3088;&#x308B;&#x6CBB;&#x7642;&#x3092;&#x8A55;&#x4FA1;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B;&#x3060;&#x3063;&#x305F;&#x3002;

&#x96FB;&#x6C17;&#x5316;&#x5B66;&#x7642;&#x6CD5;&#x306B;&#x3088;&#x308B;&#x72AC;&#x306E;&#x809B;&#x9580;&#x5468;&#x56F2;&#x816B;&#x760D;&#x306E;&#x52B9;&#x679C;&#x7684;&#x306A;&#x6CBB;&#x7642;&#x3002;

&#x7535;&#x5316;&#x5B66;&#x7597;&#x6CD5;&#x662F;&#x5229;&#x7528;&#x672C;&#x5730;&#x4F20;&#x9012;&#x3001; &#x77ED;&#x6FC0;&#x70C8;&#x7684;&#x76F4;&#x6D41;&#x7535;&#x6D41;&#x7535;&#x8109;&#x51B2;&#x5BF9;&#x80BF;&#x7624;&#x7ED3;&#x6838;&#x5316;&#x7597;&#x7684;&#x80BF;&#x7624;&#x6CBB;&#x7597;&#x3002;&#x672C;&#x7814;&#x7A76;&#x7684;&#x76EE;&#x7684;&#x662F;&#x8BC4;&#x4EF7;&#x72AC;&#x4E0D;&#x540C;&#x5927;&#x5C0F;&#x7684;&#x809B;&#x5468;&#x80BF;&#x7624;&#x7535;&#x5316;&#x5B66;&#x6CBB;&#x7597;&#x3002;

&#x6709;&#x6548;&#x6CBB;&#x7597;&#x809B;&#x5468;&#x80BF;&#x7624;&#x7535;&#x5316;&#x5B66;&#x6CBB;&#x7597;&#x72AC;&#x3002;

Electrochemotherapy is an antitumor therapy that utilizes locally-delivered, short intense direct current electric pulse to the tumor nodule plus chemotherapy. The aim of the present study was to evaluate the electrochemotherapy treatment of perianal tumors of different sizes in dogs.

Effective treatment of perianal tumors in dogs with electrochemotherapy.

Electrochemotherapy est une th&eacute;rapie antitumorale qui utilise le courant intense offerts localement, courte impulsion &eacute;lectrique au nodule de tumeur plus chimioth&eacute;rapie. Le but de cette &eacute;tude &eacute;tait d&#39;&eacute;valuer le traitement electrochemotherapy des tumeurs p&eacute;rianales de diff&eacute;rentes tailles chez les chiens.

Traitement efficace des tumeurs p&eacute;rianales chez les chiens atteints d&#39;electrochemotherapy.

&#x8FD9;&#x9879;&#x7814;&#x7A76;&#x7684;&#x76EE;&#x7684;&#x662F;&#x624B;&#x672F;&#x7684;&#x8BC4;&#x4F30;&#x4E0E;&#x987A;&#x94C2;&#x7684;&#x7535;&#x5316;&#x5B66; (ECT) &#x6CBB;&#x7597;&#x5C40;&#x90E8;&#x6CBB;&#x7597;&#x7684;&#x6210;&#x6548;&#xFF0C;&#x5E76;&#x4E0E;&#x80A5;&#x5927;&#x7EC6;&#x80DE;&#x80BF;&#x7624; (MCT) &#x7684;&#x72D7;&#x6CBB;&#x7597;&#x6548;&#x679C;&#x8FDB;&#x884C;&#x6BD4;&#x8F83;&#x3002;

&#x7535;&#x5316;&#x5B66;&#x6CBB;&#x7597;&#x72AC;&#x80A5;&#x5927;&#x7EC6;&#x80DE;&#x80BF;&#x7624;&#x7684;&#x624B;&#x672F;&#x6CBB;&#x7597;&#x76F8;&#x6BD4;&#x3002;

The aim of this study was to evaluate the effectiveness of local treatment electrochemotherapy (ECT) with cisplatin and to compare it with effectiveness of surgery for treatment of mast cell tumours (MCT) in dogs.

Electrochemotherapy compared to surgery for treatment of canine mast cell tumours.

Le but de cette &eacute;tude &eacute;tait d&#39;&eacute;valuer l&#39;efficacit&eacute; du traitement local electrochemotherapy (ECT) avec le cisplatine et de la comparer avec l&#39;efficacit&eacute; de la chirurgie pour le traitement des tumeurs des cellules de m&acirc;t (MCT) chez le chien.

Electrochemotherapy par rapport &agrave; la chirurgie pour le traitement des tumeurs des cellules de m&acirc;t canine.

Ziel dieser Studie war, die Wirksamkeit der lokalen Behandlung Electrochemotherapy (ECT) mit Cisplatin bewerten und vergleichen es mit der Wirksamkeit der Chirurgie zur Behandlung von Mastzelle Tumoren (MCT) bei Hunden.

Electrochemotherapy im Vergleich zu Chirurgie f&uuml;r die Behandlung von eckzahn Mastzelle Tumoren.

Lo scopo di questo studio era di valutare l&#39;efficacia del trattamento locale electrochemotherapy (ECT) con cisplatino e di confrontarlo con l&#39;efficacia di un intervento chirurgico per il trattamento dei tumori delle cellule dell&#39;albero (MCT) in cani.

Electrochemotherapy rispetto alla chirurgia per il trattamento di tumori canini mast cell.

&#x672C;&#x7814;&#x7A76;&#x306E;&#x76EE;&#x7684;&#x306F;&#x5C40;&#x6240;&#x6CBB;&#x7642;&#x3067;&#x96FB;&#x6C17;&#x5316;&#x5B66;&#x7642;&#x6CD5; &#xFF08;ECT&#xFF09; &#x3068;&#x30B7;&#x30B9;&#x30D7;&#x30E9;&#x30C1;&#x30F3;&#x4F75;&#x7528;&#x7642;&#x6CD5;&#x306E;&#x6709;&#x52B9;&#x6027;&#x3092;&#x8A55;&#x4FA1;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B;&#x3001;&#x72AC;&#x306E;&#x80A5;&#x6E80;&#x7D30;&#x80DE;&#x816B; (MCT) &#x306E;&#x6CBB;&#x7642;&#x306E;&#x305F;&#x3081;&#x624B;&#x8853;&#x306E;&#x6709;&#x52B9;&#x6027;&#x3092;&#x6BD4;&#x8F03;&#x3059;&#x308B;.

&#x96FB;&#x6C17;&#x5316;&#x5B66;&#x7642;&#x6CD5;&#x306B;&#x3088;&#x308B;&#x72AC;&#x80A5;&#x6E80;&#x7D30;&#x80DE;&#x816B;&#x306E;&#x6CBB;&#x7642;&#x306E;&#x305F;&#x3081;&#x624B;&#x8853;&#x3068;&#x6BD4;&#x8F03;&#x3057;&#x3066;&#x3002;

&#xC774; &#xC5F0;&#xAD6C;&#xC758; &#xBAA9;&#xC801;&#xC740; cisplatin&#xC73C;&#xB85C; &#xB85C;&#xCEEC; &#xCE58;&#xB8CC; electrochemotherapy (&#xC694;&#xBC95;)&#xC758; &#xD6A8;&#xACFC;&#xB97C; &#xD3C9;&#xAC00; &#xD558; &#xACE0; &#xAC1C;&#xC5D0;&#xC11C; &#xB3DB;&#xB300; &#xC138;&#xD3EC; &#xC885;&#xC591; (MCT)&#xC758; &#xCE58;&#xB8CC;&#xB97C; &#xC704;&#xD574; &#xC218;&#xC220;&#xC758; &#xD6A8;&#xACFC;&#xC640; &#xBE44;&#xAD50; &#xD558;&#xB294;.

&#xAC1C;&#xACFC;&#xC758; &#xB9C8;&#xC2A4;&#xD2B8; &#xC138;&#xD3EC; &#xC885;&#xC591;&#xC758; &#xCE58;&#xB8CC;&#xB97C; &#xC704;&#xD574; &#xC218;&#xC220;&#xC5D0; &#xBE44;&#xD574; Electrochemotherapy.

El objetivo de este estudio fue evaluar la efectividad del tratamiento local electrochemotherapy (ECT) con cisplatino y compararla con la efectividad de la cirug&iacute;a para el tratamiento de tumores de la c&eacute;lula del m&aacute;stil (MCT) en perros.

Electrochemotherapy en comparaci&oacute;n con la cirug&iacute;a para el tratamiento de tumores de caninos mastocitos.

&#x8F6F;&#x7EC4;&#x7EC7;&#x8089;&#x7624;&#x662F;&#x6311;&#x6218;&#x6210;&#x529F;&#x6CBB;&#x7597;&#x4E0E;&#x5E38;&#x89C4;&#x6CBB;&#x7597;&#x65B9;&#x6CD5;&#xFF0C;&#x56E0;&#x6B64;&#xFF0C;&#x8F83;&#x65B0;&#x7684;&#x6CBB;&#x7597;&#x65B9;&#x6CD5;&#x88AB;&#x8BA4;&#x4E3A;&#x3002;&#x5728;&#x8FD9;&#x9879;&#x7814;&#x7A76;&#xFF0C;&#x6211;&#x4EEC;&#x8BC4;&#x4EF7;&#x7EA4;&#x7EF4;&#x8089;&#x7624; SA 1 &#x5C0F;&#x9F20;&#x767D;&#x7EC6;&#x80DE;&#x4ECB;&#x7D20;-12 electrogene &#x6CBB;&#x7597; (EGT) &#x7684;&#x6297;&#x80BF;&#x7624;&#x6548;&#x679C;&#x3002;&#x6CBB;&#x7597;&#x8D28;&#x7C92;&#x5DF2;&#x6CE8;&#x5165;&#x6216;&#x8005;&#x4F53;&#x5185;&#xFF0C;&#x5230;&#x76AE;&#x4E0B; SA 1 &#x7ED3;&#x8282;&#x6216; intradermally &#x7624;&#x5468;&#x533A;&#x57DF;&#x3002;&#x5355;&#x4E00;&#x7684;&#x8D28;&#x7C92; DNA &#x7684;&#x5E94;&#x7528;&#xFF0C;&#x4E0E;&#x91CD;&#x5927;&#x7624;&#x5185;&#x548C; IL-12 &#x548C; IFNgamma &#x7684;&#x7CFB;&#x7EDF;&#x751F;&#x4EA7;&#x540E;&#xFF0C;&#x6211;&#x4EEC;&#x53D6;&#x5F97;&#x4E86;&#x5C40;&#x90E8;&#x548C;&#x5168;&#x8EAB;&#x6297;&#x80BF;&#x7624;&#x6548;&#x679C;&#x663E;&#x8457;&#x4E0E;&#x8FD9;&#x4E24;&#x79CD;&#x65B9;&#x6CD5;&#x3002;&#x7624;&#x5185;&#x767D;&#x7EC6;&#x80DE;&#x4ECB;&#x7D20;-12 EGT &#x5BFC;&#x81F4; 90%&#x4EE5;&#x4E0A;&#x6709; 60%&#x7684;&#x6297;&#x6027;&#x6311;&#x6218;&#x4E0E; SA 1 &#x80BF;&#x7624;&#x7EC6;&#x80DE;&#x88AB;&#x6CBB;&#x6108;&#x5C0F;&#x9F20;&#x6CBB;&#x7597;&#x80BF;&#x7624;&#x7684;&#x5B8C;&#x6574;&#x7684;&#x54CD;&#x5E94;&#x7387;&#x3002;&#x7624;&#x5468; EGT &#x5BFC;&#x81F4;&#x8F83;&#x4F4E;&#x7684;&#x5B8C;&#x6574;&#x7684;&#x54CD;&#x5E94;&#x7387; &#xFF08;16%&#xFF09;&#xFF0C;&#x5176;&#x4F59;&#x80BF;&#x7624;&#x7684;&#x663E;&#x8457;&#x589E;&#x957F;&#x6EDE;&#x3002;&#x8FD9;&#x4E24;&#x79CD;&#x7597;&#x6CD5;&#x8FD8;&#x5BFC;&#x81F4;&#x663E;&#x8457;&#x6291;&#x5236;&#x7684;&#x672A;&#x7ECF;&#x5904;&#x7406;&#x7684;&#x80BF;&#x7624;&#xFF0C;&#x5728;&#x8FDC;&#x5904;&#x7AD9;&#x70B9;&#x540C;&#x65F6;&#x65E5;&#x76CA;&#x589E;&#x957F;&#x3002;&#x8FD9;&#x4E9B;&#x6570;&#x636E;&#x8868;&#x660E;&#x767D;&#x7EC6;&#x80DE;&#x4ECB;&#x7D20;-12 EGT &#x53EF;&#x80FD;&#x6709;&#x7528;&#x7684;&#x8F6F;&#x7EC4;&#x7EC7;&#x8089;&#x7624;&#x6CBB;&#x7597;&#x4E2D;&#x53D1;&#x6325;&#x5C40;&#x90E8;&#x548C;&#x5168;&#x8EAB;&#x7684;&#x6297;&#x80BF;&#x7624;&#x4F5C;&#x7528;&#x3002;

&#x5C40;&#x90E8;&#x548C;&#x5168;&#x8EAB;&#x6297;&#x80BF;&#x7624;&#x7624;&#x5185;&#x548C;&#x7624;&#x5468;&#x767D;&#x7EC6;&#x80DE;&#x4ECB;&#x7D20;-12 electrogene &#x6CBB;&#x7597;&#x5BF9;&#x5C0F;&#x9F20;&#x8089;&#x7624;&#x3002;

Soft tissue sarcomas pose a challenge for successful treatment with conventional therapeutic methods, therefore newer therapeutic approaches are considered. In this study, we evaluated the antitumor effect of IL-12 electrogene therapy (EGT) on murine SA-1 fibrosarcoma. The therapeutic plasmid was injected either intratumorally into subcutaneous SA-1 nodules or intradermally into the peritumoral region. We achieved a remarkable local and systemic antitumor effect with both approaches after single plasmid DNA application, with significant intratumoral and systemic production of IL-12 and IFNgamma. Intratumoral IL-12 EGT resulted in over 90% complete response rate of the treated tumors with 60% of cured mice being resistant to challenge with SA-1 tumor cells. Peritumoral EGT resulted in a lower complete response rate (16%), with significant growth delay of remaining tumors. Both therapies also resulted in significant inhibition of growth of untreated tumors, growing simultaneously at a distant site. These data suggest that IL-12 EGT may be useful in the treatment of soft tissue sarcomas, exerting a local and systemic antitumor effect.

Local and systemic antitumor effect of intratumoral and peritumoral IL-12 electrogene therapy on murine sarcoma.

Sarcomes des tissus mous sont un d&eacute;fi pour la r&eacute;ussite du traitement avec les m&eacute;thodes th&eacute;rapeutiques conventionnelles, par cons&eacute;quent, des approches th&eacute;rapeutiques nouvelles sont consid&eacute;r&eacute;s. Dans cette &eacute;tude, nous avons &eacute;valu&eacute; l&#39;effet antitumoral de IL-12 ont th&eacute;rapie (EGT) sur murine Fibrosarcome SA-1. Le plasmide th&eacute;rapeutique a &eacute;t&eacute; inject&eacute; soit intratumorally dans les nodules sous-cutan&eacute;s SA-1, ou par voie intradermique dans la r&eacute;gion de p&eacute;ri-tumoral. Nous avons atteint un remarquable effet antitumoral local et syst&eacute;mique avec les deux approches apr&egrave;s application d&#39;ADN de plasmide, avec intratumorale importante et la production d&#39;IL-12 et IFNgamma syst&eacute;mique. EGT intratumorale IL-12 a donn&eacute; lieu &agrave; plus de 90 % des taux de r&eacute;ponse compl&egrave;te des tumeurs trait&eacute;es avec 60 % des souris polym&eacute;ris&eacute;es r&eacute;sistantes &agrave; la provocation par des cellules tumorales SA-1. P&eacute;ritumoral EGT aboutit &agrave; un plus faible taux de r&eacute;ponse compl&egrave;te (16 %), avec un retard de croissance significative des tumeurs restantes. Les deux traitements ont &eacute;galement entra&icirc;n&eacute; une inhibition significative de la croissance des tumeurs non trait&eacute;es, de plus en plus en m&ecirc;me temps sur un site distant. Ces donn&eacute;es sugg&egrave;rent que EGT d&#39;IL-12 peut &ecirc;tre utile dans le traitement des sarcomes des tissus mous, exer&ccedil;ant un effet antitumoral local et syst&eacute;mique.

Locale et syst&eacute;mique effet antitumoral intratumorale et p&eacute;ri-tumoral IL-12 ont th&eacute;rapie sur sarcome murin.

Weichteilsarkomen eine Herausforderung f&uuml;r eine erfolgreiche Behandlung mit konventionellen Behandlungsmethoden darstellen, gelten daher als neuere therapeutische Ans&auml;tze. In dieser Studie haben wir die antitumor-Wirkung von IL-12 Electrogene Therapie (EGT) auf murinen SA-1 Fibrosarkom ausgewertet. Die therapeutische Plasmid wurde entweder Intratumorally in subkutane Kn&ouml;tchen der SA-1 oder intradermal in der Region Peritumorales eingespritzt. Wir erzielten einen bemerkenswerten lokalen und systemischen antitumor-Effekt mit beiden Ans&auml;tzen nach einzelnen Plasmid DNA-Anwendung mit erheblichen intratumorale und systemische Produktion von IL-12 und IFNgamma. Intratumorale IL-12 EGT f&uuml;hrte zu &uuml;ber 90 % komplette Ansprechrate der behandelten Tumoren mit 60 % des ausgeh&auml;rteten M&auml;use Wesen best&auml;ndig Herausforderung mit SA-1-Tumorzellen. Peritumorales EGT ergab eine niedrigere Rate der komplette Remission (16 %), mit bedeutende Wachstumsverz&ouml;gerung der &uuml;brigen Tumoren. Beide Therapien f&uuml;hrte ebenfalls signifikante Hemmung des Wachstums von unbehandelten Tumoren, w&auml;chst gleichzeitig an einem entfernten Standort. Diese Daten legen nahe, dass IL-12 EGT n&uuml;tzlich bei der Behandlung von Weichteilsarkomen, m&ouml;glicherweise eine lokale und systemische antitumor-Wirkung auszu&uuml;ben.

Lokale und systemische antitumor Effekt intratumorale und Peritumorales IL-12 Electrogene Therapie auf murinen Sarkom.

Sarcomi dei tessuti molli rappresentano una sfida per il successo del trattamento con metodi terapeutici convenzionali, pertanto sono considerati pi&ugrave; recenti approcci terapeutici. In questo studio abbiamo valutato l&#39;effetto antitumorale della terapia di electrogene IL-12 (EGT) su fibrosarcoma murino di SA-1. Il plasmide terapeutico &egrave; stato iniettato o intratumorally in noduli sottocutanei SA-1, o per via intradermica nella regione peritumoral. Abbiamo ottenuto un notevole effetto antitumorale locale e sistemico con entrambi gli approcci dopo l&#39;applicazione di DNA del plasmide singolo, con significativo intratumoral e produzione sistemica di IL-12 e IFN. IL Intratumoral-12 EGT portato in oltre il 90% tasso di risposta completa dei tumori trattati con il 60% dei topi curati essendo resistenti alla sfida con cellule tumorali SA-1. Peritumoral EGT ha portato in un basso tasso di risposta completa (16%), con un ritardo di crescita significativa di tumori rimanenti. Entrambe le terapie ha portato anche in inibizione significativa della crescita dei tumori non trattati, cresce simultaneamente in un sito distante. Questi dati suggeriscono che IL-12 EGT pu&ograve; essere utile nel trattamento dei sarcomi dei tessuti molli, esercitando un effetto antitumorale locale e sistemico.

Locale e sistemico effetto antitumorale della terapia di electrogene IL-12 intratumoral e peritumoral sul sarcoma murino.

&#x8EDF;&#x90E8;&#x7D44;&#x7E54;&#x8089;&#x816B;&#x306F;, &#x5F93;&#x6765;&#x306E;&#x6CBB;&#x7642;&#x6CD5;&#x3068;&#x6CBB;&#x7642;&#x6210;&#x529F;&#x306E;&#x305F;&#x3081;&#x306E;&#x6311;&#x6226;&#x30DD;&#x30FC;&#x30BA;&#x3001;&#x3057;&#x305F;&#x304C;&#x3063;&#x3066;&#x65B0;&#x3057;&#x3044;&#x6CBB;&#x7642;&#x306E;&#x30A2;&#x30D7;&#x30ED;&#x30FC;&#x30C1;&#x3068;&#x898B;&#x306A;&#x3055;&#x308C;&#x307E;&#x3059;&#x3002;&#x672C;&#x7814;&#x7A76;&#x3067;&#x306F;, &#x30DE;&#x30A6;&#x30B9;&#x306E; SA 1 &#x7DDA;&#x7DAD;&#x8089;&#x816B;&#x3067; IL 12 electrogene &#x7642;&#x6CD5; (EGT) &#x306E;&#x6297;&#x816B;&#x760D;&#x52B9;&#x679C;&#x3092;&#x8A55;&#x4FA1;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x6CBB;&#x7642;&#x306E;&#x30D7;&#x30E9;&#x30B9;&#x30DF;&#x30C9;&#x3069;&#x3061;&#x3089; intratumorally &#x76AE;&#x4E0B; SA 1 &#x7D50;&#x7BC0;&#x3084;&#x63BB;&#x304D;&#x816B;&#x760D;&#x5468;&#x56F2;&#x9818;&#x57DF;&#x306B;&#x6CE8;&#x5165;&#x3055;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;&#x5358;&#x4E00;&#x306E;&#x30D7;&#x30E9;&#x30B9;&#x30DF;&#x30C9; DNA &#x30A2;&#x30D7;&#x30EA;&#x30B1;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x306B;&#x91CD;&#x8981;&#x306A;&#x816B;&#x760D;&#x3068;&#x5168;&#x8EAB;&#x751F;&#x7523; il-12 &#x3068; IFNgamma &#x306E;&#x5F8C;&#x3001;&#x9855;&#x8457;&#x306A;&#x5168;&#x8EAB;&#x53CA;&#x3073;&#x80BA;&#x5C40;&#x6240;&#x6297;&#x816B;&#x760D;&#x52B9;&#x679C;&#x306E;&#x4E21;&#x65B9;&#x306E;&#x30A2;&#x30D7;&#x30ED;&#x30FC;&#x30C1;&#x3092;&#x9054;&#x6210;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x816B;&#x760D;&#x5185; IL 12 EGT &#x7D50;&#x679C;&#x3067; 90% &#x4EE5;&#x4E0A;&#x6271;&#x308F;&#x308C;&#x305F;&#x816B;&#x760D;&#x306E;&#x5B8C;&#x5168;&#x594F;&#x52B9;&#x7387; 60 &#xFF05; &#x306E;&#x6CBB;&#x7652;&#x3057;&#x305F;&#x30DE;&#x30A6;&#x30B9;&#x306B;&#x5BFE;&#x3057;&#x3066;&#x62B5;&#x6297;&#x3059;&#x308B;&#x30C1;&#x30E3;&#x30EC;&#x30F3;&#x30B8; SA 1 &#x816B;&#x760D;&#x7D30;&#x80DE;&#x3092;&#x3002;&#x816B;&#x760D;&#x5468;&#x56F2; EGT &#x306F;&#x4F4E;&#x3044;&#x5B8C;&#x5168;&#x594F;&#x52B9;&#x7387; &#xFF08;16 &#xFF05;&#xFF09;&#x3001;&#x6B8B;&#x308A;&#x306E;&#x816B;&#x760D;&#x306E;&#x5927;&#x5E45;&#x306A;&#x6210;&#x9577;&#x9045;&#x5EF6;&#x3092;&#x3082;&#x305F;&#x3089;&#x3057;&#x305F;&#x3002;&#x4E21;&#x65B9;&#x306E;&#x6CBB;&#x7642;&#x3082;&#x540C;&#x6642;&#x306B;&#x9060;&#x3044;&#x30B5;&#x30A4;&#x30C8;&#x306B;&#x6210;&#x9577;&#x3057;&#x3066;&#x3001;&#x672A;&#x51E6;&#x7406;&#x306E;&#x816B;&#x760D;&#x306E;&#x6210;&#x9577;&#x3092;&#x6709;&#x610F;&#x306B;&#x963B;&#x5BB3;&#x306E;&#x7D50;&#x679C;&#x3002;&#x3053;&#x308C;&#x3089;&#x306E;&#x30C7;&#x30FC;&#x30BF; IL 12 EGT &#x5168;&#x8EAB;&#x53CA;&#x3073;&#x80BA;&#x5C40;&#x6240;&#x6297;&#x816B;&#x760D;&#x52B9;&#x679C;&#x3092;&#x767A;&#x63EE;&#x3001;&#x8EDF;&#x90E8;&#x7D44;&#x7E54;&#x8089;&#x816B;&#x306E;&#x6CBB;&#x7642;&#x306B;&#x6709;&#x7528;&#x306A;&#x3053;&#x3068;&#x304C;&#x793A;&#x5506;&#x3055;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;

&#x5168;&#x8EAB;&#x53CA;&#x3073;&#x80BA;&#x5C40;&#x6240;&#x30DE;&#x30A6;&#x30B9;&#x8089;&#x816B;&#x306E;&#x6297;&#x816B;&#x760D;&#x52B9;&#x679C;&#x306E;&#x816B;&#x760D;&#x3068;&#x816B;&#x760D;&#x5468;&#x56F2;&#x306E; IL-12 electrogene &#x7642;&#x6CD5;&#x3002;

&#xC5F0; &#xC870;&#xC9C1; sarcomas &#xAE30;&#xC874;&#xC758; &#xCE58;&#xB8CC; &#xBC29;&#xBC95;&#xC73C;&#xB85C; &#xC131;&#xACF5;&#xC801;&#xC778; &#xCE58;&#xB8CC;&#xC5D0; &#xB300; &#xD55C; &#xB3C4;&#xC804; &#xD3EC;&#xC988;, &#xB530;&#xB77C;&#xC11C; &#xC0C8;&#xB85C;&#xC6B4; &#xCE58;&#xB8CC; &#xC811;&#xADFC;&#xC774; &#xAC04;&#xC8FC; &#xB429;&#xB2C8;&#xB2E4;. &#xC774; &#xC5F0;&#xAD6C;&#xC5D0;&#xC11C; &#xC6B0;&#xB9AC;&#xAC00; murine SA 1 fibrosarcoma antitumor IL-12 electrogene &#xC694;&#xBC95; (EGT) &#xD6A8;&#xACFC; &#xD3C9;&#xAC00;. &#xD53C;&#xD558; SA 1 &#xD639; &#xB610;&#xB294; intradermally peritumoral &#xC9C0;&#xC5ED;&#xC5D0; &#xCE58;&#xB8CC; &#xD50C;&#xB77C;&#xC2A4; &#xBBF8;&#xB4DC; &#xC911; intratumorally&#xB294; &#xC8FC;&#xC785; &#xD588;&#xB2E4;. &#xC6B0;&#xB9AC;&#xB294; &#xB2E8;&#xC77C; &#xD50C;&#xB77C;&#xC2A4; &#xBBF8;&#xB4DC; DNA &#xC751;&#xC6A9; &#xD504;&#xB85C;&#xADF8;&#xB7A8;&#xC744; &#xC911;&#xC694; &#xD55C; intratumoral &#xBC0F; IL-12&#xC640; Ifngamma&#xC758; &#xCCB4;&#xACC4;&#xC801;&#xC778; &#xC0DD;&#xC0B0; &#xD6C4; &#xB180;&#xB77C;&#xC6B4; &#xB85C;&#xCEEC; &#xBC0F; &#xC870;&#xC9C1;&#xC758; antitumor &#xD6A8;&#xACFC; &#xB450; &#xAC00;&#xC9C0; &#xBC29;&#xBC95; &#xBAA8;&#xB450; &#xD568;&#xAED8; &#xB2EC;&#xC131;. Intratumoral IL-12 EGT &#xACB0;&#xACFC; 90% &#xC774;&#xC0C1; &#xCE58;&#xC720; &#xC950; &#xC800;&#xD56D; SA 1 &#xC885;&#xC591; &#xC138;&#xD3EC;&#xC5D0; &#xB300; &#xD55C; &#xB3C4;&#xC804;&#xC758; 60% &#xCE58;&#xB8CC; &#xC885;&#xC591;&#xC758; &#xC644;&#xC804; &#xD55C; &#xC751;&#xB2F5; &#xC18D;&#xB3C4;. Peritumoral EGT &#xB0AE;&#xC740; &#xC644;&#xC804; &#xD55C; &#xC751;&#xB2F5; &#xC18D;&#xB3C4; (16%), &#xB0A8;&#xC544; &#xC788;&#xB294; &#xC885;&#xC591;&#xC758; &#xC911;&#xC694; &#xD55C; &#xC131;&#xC7A5; &#xC9C0;&#xC5F0; &#xACB0;&#xACFC;. &#xB450; &#xC694;&#xBC95; &#xB610;&#xD55C; &#xC0C1;&#xB2F9;&#xD55C; &#xBA3C; &#xC0AC;&#xC774;&#xD2B8;&#xC5D0; &#xB3D9;&#xC2DC;&#xC5D0; &#xC131;&#xC7A5; &#xCE58;&#xB8CC; &#xC885;&#xC591;&#xC758; &#xC131;&#xC7A5; &#xC5B5;&#xC81C; &#xADC0;&#xCC29;&#xB418; &#xC5C8; &#xB2E4;. &#xC774;&#xB7EC;&#xD55C; &#xB370;&#xC774;&#xD130; IL-12 EGT &#xAC83;&#xC740; &#xBD80;&#xB4DC;&#xB7EC;&#xC6B4; &#xC870;&#xC9C1; sarcomas, &#xCE58;&#xB8CC;&#xC5D0; &#xC720;&#xC6A9; &#xD558; &#xAC8C; &#xB85C;&#xCEEC; &#xC870;&#xC9C1;&#xC758; antitumor &#xD6A8;&#xACFC; &#xBC1C;&#xD718; &#xD558;&#xB294; &#xAC83;&#xC774; &#xC88B;&#xC2B5;&#xB2C8;&#xB2E4;.

&#xB85C;&#xCEEC; &#xBC0F; &#xC870;&#xC9C1;&#xC758; antitumor &#xD6A8;&#xACFC;&#xC758; intratumoral &#xBC0F; peritumoral IL-12 electrogene &#xCE58;&#xB8CC; murine &#xC721; &#xC885;&#xC5D0;.

Sarcomas de partes blandas representan un desaf&iacute;o para un tratamiento exitoso con m&eacute;todos terap&eacute;uticos convencionales, por lo tanto m&aacute;s nuevos enfoques terap&eacute;uticos se consideran. En este estudio, se evalu&oacute; el efecto antitumoral de la IL-12 aggregaat terapia (EGT) en fibrosarcoma murino de SA-1. El pl&aacute;smido terap&eacute;utico fue inyectado o intratumorally en n&oacute;dulos subcut&aacute;neos de SA-1 o por v&iacute;a intrad&eacute;rmica en la regi&oacute;n de peritumoral. Hemos conseguido un notable efecto antitumoral local y sist&eacute;mico con ambos enfoques despu&eacute;s de la aplicaci&oacute;n de ADN pl&aacute;smido sola, con significativa intratumoral y sist&eacute;mica producci&oacute;n de IL-12 y IFNgamma. EGT intratumoral IL-12 result&oacute; en m&aacute;s del 90% la tasa de respuesta completa de los tumores tratados con 60% de los ratones curados ser resistentes al desaf&iacute;o con las c&eacute;lulas del tumor SA-1. Peritumoral EGT result&oacute; en una menor tasa de respuesta completa (16%), con retraso en el crecimiento significativo de tumores restantes. Ambas terapias tambi&eacute;n produjo una inhibici&oacute;n significativa del crecimiento de tumores no tratados, creciendo al mismo tiempo en un sitio distante. Estos datos sugieren que IL-12 EGT puede ser &uacute;til en el tratamiento de los sarcomas de partes blandas, ejerciendo un efecto antitumoral local y sist&eacute;mico.

Local y sist&eacute;mico efecto antitumoral de la terapia de aggregaat de IL-12 intratumoral y peritumoral en sarcoma murino.

Laboratory and clinical parameters of 149 dogs, exposed to Anaplasma phagocytophilum (A. phagocytophilum), and 19 control dogs were evaluated and compared retrospectively. The aim of our study was to determine statistically significant differences of selected parameters between groups of patients, divided according to the immunofluorescence (IFA) titres, in attempt to improve current diagnostic and treatment criteria. Exposure to A. phagocytophilum was confirmed by IFA and infection by PCR. Based on the results, the dogs were divided into 8 groups (6 groups of seropositive dogs according to the antibody titre, 1 group of PCR positive dogs, and a control group). Selected parameters were compared between groups. Thrombocytopenia was confirmed to be the most prominent haematological change in IFA and/or PCR positive dogs. There were no statistically significant differences in clinical and haematological observations between groups of different IFA titre but clear overall differences between each IFA and PCR positive groups compared to the control group. Our results showed the necessity of introducing additional diagnostic procedures in clinical practice, since antibody titre and haematological parameters are not sufficient to confirm the clinical relevance of exposure to A. phagocytophilum in a particular patient.

Anaplasmosis in dogs: the relation of haematological, biochemical and clinical alterations to antibody titre and PCR confirmed infection.

Mast cell tumors (MCT) are the most common malignant cutaneous tumors in dogs with extremely variable biological behaviour. Different treatment approaches can be used in canine cutaneous MCT, with surgical excision being the treatment of choice. In this study, electrogene therapy (EGT) as a new therapeutic approach to canine MCTs, was established. MATERIALS AND METHODS.: Eight dogs with a total of eleven cutaneous MCTs were treated with intratumoral EGT using DNA plasmid encoding human interleukin-12 (IL-12). The local response to the therapy was evaluated by repeated measurements of tumor size and histological examination of treated tumors. A possible systemic response was assessed by determination of IL-12 and interferon- γ (IFN-γ) in patients' sera. The occurence of side effects was monitored with weekly clinical examinations of treated animals and by performing basic bloodwork, consisting of the complete bloodcount and determination of selected biochemistry parameters.

Electrogene therapy with interleukin-12 in canine mast cell tumors.

Human immunodeficiency virus (HIV) is associated with several renal syndromes including acute and chronic renal failures, but the underlying pathogenic mechanisms are unclear. HIV and feline immunodeficiency virus (FIV) share numerous biological and pathological features, including renal alterations. We investigated and compared the morphological changes of renal tissue of 51 experimentally and 21 naturally infected cats. Compared to the latter, the experimentally infected cats exhibited some mesangial widening and glomerulonephritis, milder proteinuria, and lower tubular and interstitial alterations. The numbers of giant protein tubular casts and tubular microcysts were also lower. In contrast, diffuse interstitial infiltrates and glomerular and interstitial amyloidosis were detected only in naturally infected cats. Similar alterations are found in HIV infected patients, thus supporting the idea of a causative role of FIV infection in renal disease, and underlining the relevance of the FIV and its natural host as an animal model for investigating lentivirus-associated nephropathy.

Renal alterations in feline immunodeficiency virus (FIV)-infected cats: a natural model of lentivirus-induced renal disease changes.

The use of large animals as an experimental model for novel treatment techniques has many advantages over the use of laboratory animals, so veterinary medicine is becoming an increasingly important translational bridge between preclinical studies and human medicine. The results of preclinical studies show that gene therapy with therapeutic gene encoding interleukin-12 (IL-12) displays pronounced antitumor effects in various tumor models. A number of different studies employing this therapeutic plasmid, delivered by either viral or non-viral methods, have also been undertaken in veterinary oncology. In cats, adenoviral delivery into soft tissue sarcomas has been employed. In horses, naked plasmid DNA has been delivered by direct intratumoral injection into nodules of metastatic melanoma. In dogs, various types of tumors have been treated with either local or systemic IL-12 electrogene therapy. The results of these studies show that IL-12 based gene therapy elicits a good antitumor effect on spontaneously occurring tumors in large animals, while being safe and well tolerated by the animals. Hopefully, such results will lead to further investigation of this therapy in veterinary medicine and successful translation into human clinical trials.

IL-12 based gene therapy in veterinary medicine.

The aim of this study was to evaluate the efficacy and safety of electrochemotherapy (ECT) with bleomycin for treatment of squamous cell carcinoma (SCC) in cats. Between March 2008 and October 2011, 11 cats with 17 superficial SCC nodules in different clinical stages (ranging from Tis to T4), located on nasal planum (6/11), pinnae (3/11) and both locations (2/11), were included in a prospective non-randomised study. Sixteen of 17 SCC nodules were treated with ECT (15/16 with single session and in one case with two sessions); one nodule was surgically removed. Altogether, complete response (CR) was achieved for 81.8% (9/11) cats and 87.5% (14/16) nodules, lasting from 2 months up to longer than 3 years. Only 2/9 cats in which CR was initially observed, had recurrence 2 and 8 months after the ECT procedure. In the remaining two cats with highly infiltrative spread into adjacent tissues, progression of the disease was observed, despite ECT, and both were euthanased 4 and 5 months after the procedure. ECT in cats was well tolerated and no evident local or systemic side effects were observed. The results of this study suggest that ECT is a highly effective and safe method of local tumour control of feline cutaneous SCCs. It should be considered as an alternative treatment option, especially when other treatment approaches are not acceptable by the owners, owing to their invasiveness, mutilation or high cost.

Electrochemotherapy with intravenous bleomycin injection: an observational study in superficial squamous cell carcinoma in cats.

Alterations in serum protein profile, presence of circulating immune complexes (CIC) and proteinuria were investigated in a large group of dogs naturally infected with the Anaplasma phagocytophilum bacterium. Our aim was to evaluate the presence of hypergammaglobulinaemia, CIC and proteinuria as a possible result of an immune-mediated disease following infection by or exposure to A. phagocytophilum. Dogs were divided into three groups - IFA positive (188 dogs with confirmed exposure to A. phagocytophilum), PCR positive (31 dogs with confirmed infection), and control (IFA and PCR negative) (19 dogs). Serum and urine protein patterns were determined by electrophoresis and CIC concentrations by absorbance nephelometry. No significant differences in hypergammaglobulinaemia were observed between the different groups, as shown by the presence of acute phase proteins α2 and β1-2 globulins. CIC concentrations in the IFA and PCR positive groups were, on average, higher than in controls by 151.3μg/ml, though the differences were not significant. The proportion of dogs with proteinuria did not differ significantly between groups. Our results confirm the assumption that anaplasmosis in dogs is most probably a disease with an acute course, with a good prognosis under the right treatment.

Serum protein profiles, circulating immune complexes and proteinuria in dogs naturally infected with Anaplasma phagocytophilum.

Mycoplasma (M.) cynos is a proven pathogen of dogs causing respiratory infections including pneumonia. We examined 19 M. cynos strains isolated from different organs of dogs in Austria, Denmark and Israel. All strains agglutinated mammalian and chicken erythrocytes. Using erythrocytes of chickens or dogs as specific ligands we isolated an approximately 65 kDa protein from cell-free supernatants of 3 M. cynos strains, which showed an apparent capacity for haemagglutination. The N-terminal sequence of a 25 kDa fragment of this protein was identified as NNEMTPKVTVEAKSMELLLSVEK. The identical amino acid sequence is encoded by the gene MCYN_0308 in the genome of M. cynos C142. This gene belongs to a family of some 20 genes which encode putative lipoproteins with proline-rich regions (PRR) in the first third of their molecules. We termed the 65 kDa haemagglutinin HapA and sequenced hapA gene homologues of 16 M. cynos strains. Analyses of hapA gene homologues revealed similar but not identical sequences, some having insertions and/or deletions in the PRR. We produced a recombinant HapA protein (rHapA) and also mouse monoclonal antibodies (mAbs) recognizing HapA. However, enzyme immunoassays using native M. cynos colonies and mAbs 5G2 or 3B7 showed variable expression of HapA in all M. cynos strains. This was further confirmed by Western blot analyses which showed different HapA quantities and also size-variation of HapA among strains. Analyses of cDNA of the expressed hapA genes showed that besides the hapA gene cultures of M. cynos (strains 105, 2002, 2297) can also express other forms of hap genes. In addition, in cloned cultures of strain 2297 altered HapA epitopes for mAbs 5G2 and 3B7 with distinct hapA gene mutations that resulted in altered HapA amino acid sequence were found. Most of the dogs examined had serum antibodies to rHapA. In conclusion, we characterized the M. cynos haemagglutinin HapA protein and encoding gene hapA, a factor involved in cytadherence to host cells and therefore important for M. cynos infection, and showed that expression of HapA is varied in M. cynos by two distinct mechanisms; differential gene expression and nucleic acid substitution within hapA homologues.

Molecular characterisation of the Mycoplasma cynos haemagglutinin HapA.

A gene electrotransfer (GET) of interleukin 12 (IL-12) had already given good results when treating tumors in human and veterinary clinical trials. So far, plasmids used in veterinary clinical studies encoded a human or a feline IL-12 and an ampicillin resistance gene, which is not recommended by the regulatory agencies to be used in clinical trials. Therefore, the aim of the current study was to construct the plasmid encoding a canine IL-12 with kanamycin antibiotic resistance gene that could be used in veterinary clinical oncology. The validation of the newly constructed plasmid was carried out on canine malignant melanoma cells, which have not been used in GET studies so far, and on human malignant melanoma cells. Canine and human malignant melanoma cell lines were transfected with plasmid encoding enhanced green fluorescence protein at different pulse parameter conditions to determine the transfection efficiency and cell survival. The IL-12 expression of the most suitable conditions for GET of the plasmid encoding canine IL-12 was determined at mRNA level by the qRT-PCR and at protein level with the ELISpot assay. The obtained results showed that the newly constructed plasmid encoding canine IL-12 had similar or even higher expression capacity than the plasmid encoding human IL-12. Therefore, it represents a promising therapeutic plasmid for further IL-12 gene therapy in clinical studies for spontaneous canine tumors. Additionally, it also meets the main regulatory agencies' (FDA and EMA) criteria.

Natasa Tozon

Clinic for Surgery and Small Animals,
Veterinary Faculty,
Clinic for Surgery and Small Animals, Veterinary Faculty

University of Ljubljana

Operating Procedures of the Electrochemotherapy for Treatment of Tumor in Dogs and Cats

Natasa Tozon

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Clinic for Surgery and Small Animals,Veterinary Faculty,Clinic for Surgery and Small Animals, Veterinary Faculty

University of Ljubljana

Operating Procedures of the Electrochemotherapy for Treatment of Tumor in Dogs and Cats

Clinic for Surgery and Small Animals,
Veterinary Faculty,
Clinic for Surgery and Small Animals, Veterinary Faculty