Efficacy and safety of electrochemotherapy combined with peritumoral IL-12 gene electrotransfer of canine mast cell tumours.

Electrochemotherapy combined with peritumoral interleukin-12 (IL-12) gene electrotransfer was used for treatment of mast cell tumours in 18 client-owned dogs. Local tumour control, recurrence rate, as well as safety of combined therapy were evaluated. One month after the therapy, no side effects were recorded and good local tumour control was observed with high complete responses rate which even increased during the observation period to 72%. IL-12 gene electrotransfer resulted in 78% of patients with detectable serum IFN-γ and/or IL-12 levels. In the treated tumours vascular changes as well as minimal T-lymphocytes infiltration was observed. After 1 week, the plasmid DNA was not detected intra- or peritumorally and no horizontal gene transfer was observed. In summary, our study demonstrates high antitumour efficacy of electrochemotherapy combined with IL-12 electrotransfer, which also prevented recurrences or distant metastases, as well as its safety and feasibility in treatment of canine mast cell tumours.

Electrochemotherapy as a single or adjuvant treatment to surgery of cutaneous sarcoid tumours in horses: a 31-case retrospective study.

The aim of our study was to evaluate the efficacy of electrochemotherapy (ECT) with cisplatin as a single or adjuvant treatment for sarcoids in equids. Different treatment options with different success rates were proposed. Thirty-one horses and one donkey with different clinical type, size and location of tumours were treated with ECT as a single treatment (18 animals with 52 tumour nodules) or as adjuvant treatment with marginal surgical excision (14 animals with 18 tumour nodules). In animals treated only with ECT with cisplatin, complete response was obtained in 48/52 (92.3 per cent) nodules and partial response in the other 4 nodules (7.7 per cent). In most cases, one to three sessions, only in two cases four and in one case five sessions, every 4 weeks were needed to obtain the measurable response. During the observation time, only in one case was the recurrence noted 60 months after treatment. Complete response in all 18 tumour nodules treated with surgery and adjuvant ECT was obtained and only one recurrence was noted after 14 months during the observation time. The results of this study show that ECT with cisplatin is an effective, safe, and simple local treatment of sarcoids in equids. According to the tumour size and location, single or combined treatment should be performed.

Plasma and erythrocyte glutathione peroxidase activity, serum selenium concentration, and plasma total antioxidant capacity in cats with IRIS stages I-IV chronic kidney disease.

BACKGROUND: Serum selenium concentrations and the activity of plasma glutathione peroxidase (GPx) decrease with the progression of chronic kidney disease (CKD) in human patients. Selenium is considered a limiting factor for plasma GPx synthesis. Plasma total antioxidant capacity (TAC) is decreased in CKD cats in comparison to healthy cats. HYPOTHESIS: Serum selenium concentrations and plasma and erythrocyte GPx activity in cats with CKD are lower than in healthy cats. Serum selenium concentrations, the activity of enzymes, and plasma TAC progressively decrease with the progression of kidney disease according to IRIS (International Renal Interest Society) classification. ANIMALS: Twenty-six client-owned cats in IRIS stages I-IV of CKD were compared with 19 client-owned healthy cats. METHODS: A CBC, serum biochemical profile, urinalysis, plasma and erythrocyte GPx activity, serum selenium concentration, and plasma TAC were measured in each cat. RESULTS: Cats in IRIS stage IV CKD had a significantly higher (P = .025) activity of plasma GPx (23.44 ± 6.28 U/mL) than cats in the control group (17.51 ± 3.75 U/mL). There were no significant differences in erythrocyte GPx, serum selenium concentration, and plasma TAC, either among IRIS stages I-IV CKD cats or between CKD cats and healthy cats. CONCLUSIONS AND CLINICAL IMPORTANCE: Erythrocyte GPx activity, serum selenium concentration, and plasma TAC do not change in CKD cats compared with healthy cats. Selenium is not a limiting factor in feline CKD. Increased plasma GPx activity in cats with stage IV CKD suggests induction of antioxidant defense mechanisms. Antioxidant defense systems might not be exhausted in CKD in cats.

Electrochemotherapy in veterinary oncology.

Electropermeabilization is a method that uses electric field pulses to induce an electrically mediated reorganization of the plasma membrane of cells. Electrochemotherapy combines local or systemic administration of chemotherapeutic drugs such as bleomycin or cisplatin that have poor membrane permeability with electropermeabilization by direct application of electric pulses to the tumors. Preclinical studies have demonstrated excellent antitumor effectiveness of electrochemotherapy on different animal models and various tumor types, minimal toxicity, and safety of the procedure. Based on results of preclinical studies, clinical studies were conducted in human patients, which demonstrated pronounced antitumor effectiveness of electrochemotherapy with 80-85% objective responses of the treated cutaneous and SC tumors. Clinical studies in veterinary oncology have demonstrated that electrochemotherapy is very effective in the treatment of cutaneous and SC tumors of different histologic types in cats, dogs, and horses. The results of these studies have also demonstrated approximately 80% long-lasting objective responses of tumors treated by electrochemotherapy. Primary tumors of different histologic types were treated. Electrochemotherapy in veterinary oncology has future promise to be highly effective, and could be used to treat primary or recurrent solitary or multiple cutaneous and SC tumors of different histology or as an adjuvant treatment to surgery.

Efficient electrotransfection into canine muscle.

Two different types of electroporation protocols have been developed for efficient electrotransfer of plasmid DNA into skeletal muscle of experimental animals. At first, only low voltage electric pulses have been used, but lately, a combination of high and low voltage pulses has been suggested as more efficient. Up to date, in dogs, this type of electroporation protocol has never been used for muscle targeted plasmid DNA electrotransfection. In this study, we used two different DNA plasmids, one encoding green fluorescent protein and one encoding human interleukin-12. Five different electroporation protocols were evaluated. Three of them featured different combinations of high and low voltage pulses, and two were performed with delivery of low voltage pulses only. Our study shows that combination of 1 high voltage pulse (600 V/cm, 100 mus), followed by 4 low voltage pulses (80 V/cm, 100 ms, 1 Hz) yielded in the same transfection efficiency as the standard trains of low voltage pulses. However, this protocol is performed quicker and, thus, more suitable for potential use in clinical practice. In addition, it yielded in detectable systemic expression of human interleukin-12. Electrotransfer of either of the plasmids was associated with only mild and transitory local side effects, without clinically detectable systemic side effects. The results indicate that electrotransfection is a feasible, effective, and safe method for muscle targeted gene therapy in dogs, which could have potential for clinical applications in veterinary medicine of small animals.

Pattern of myosin heavy chain isoforms in different fibre types of canine trunk and limb skeletal muscles.

The aim of this study was to determine the pattern of myosin heavy chain (MHC) isoform expressions within the muscle fibres of functionally diverse trunk and limb dog muscles using monoclonal antibodies that are specific to MHC isoforms. We found that three MHC isoforms are expressed in dog skeletal muscles. The pattern of their expressions determined the existence of 'pure' fibres, i.e. I and IIa, both expressing only one MHC isoform, and 'hybrid' fibres, i.e. I/IIa and IIa/x, that co-expressed two MHC isoforms. While the MHCI, MHCIIa and MHCI/IIa fibres corresponded to the myofibrillar ATPase type fibres I, IIA and IIC, respectively, the hybrid MHCIIa/x fibres mostly behaved like the IIDog fibre type in myofibrillar ATPase reaction as described by Latorre et al. No pure MHCIIx fibres were found. Though MHCIIa/x fibres were quite numerous, their presence varied not only within different muscles but within the same muscle of different animals as well. We suggest that the discrepancies in the classification of fibre types according to their myofibrillar ATPase activity between different studies of dog skeletal muscles are probably a consequence of the variable content of the MHCIIa and MHCIIx isoforms in the MHCIIa/x hybrid fibres. Estimating the histochemical metabolic profile of fibres we found that in all fast fibres oxidative-glycolytic metabolism prevailed, whereas in slow fibres oxidative metabolism was more pronounced.

Electrochemotherapy: potentiation of local antitumour effectiveness of cisplatin in dogs and cats.

The aim of this study was to introduce electrochemotherapy with cisplatin into veterinary medicine, where there is a need for inexpensive and effective treatment of cutaneous and subcutaneous tumours of various histological types. The response to treatment was assessed on tumour nodules in 3 cats with mammary adenocarcinoma and fibrosarcoma, and in 7 dogs with mammary adenocarcinoma, cutaneous mast cell tumour, hemangioma, hemangiosarcoma, adenocarcinoma glandulae paranalis and neurofibroma. Twenty-four tumour nodules of different size were treated; 5 with cisplatin injected intratumourally and 19 with electrochemotherapy, i.e. intratumoural administration of cisplatin followed by delivery of electric pulses to the tumour nodule. Electrochemotherapy with cisplatin had a good antitumour effect on all tumours treated. Their average size 4 weeks after treatment was also greatly reduced (0.01 cm3) compared to those treated by intratumoural cisplatin injection alone (3.0 cm3). Altogether, electrochemotherapy- treated tumours responded with 84% objective responses, whereas only one tumourpartially responded to cisplatin treatment alone. Evaluated by contingency table, the response to treatment with electrochemotherapy was significantly better than that of the cisplatin treated group (p=0.014). Furthermore, there was a significant prolongation of the duration of response in electrochemotherapy treated tumours (p = 0.046). This study showed that electrochemotherapy with cisplatin is an effective, safe and simple local treatment of different histological types of cutaneous and subcutaneous tumours in cats and dogs.

Treatment of adult asthma: controlled double-blind clinical trial of oxitropium bromide.

Sixteen young adult sufferers from extrinsic paroxysmal asthma with pollen hypersensitivity took part in a therapeutic trial of the synthetic anticholinergic agent oxitropium bromide administered by a metered dose inhaler. The study comprised three 3-week periods. The first, run-in period was carried out to confirm the ability of the patients to maintain a daily record of symptoms. During the second and third periods, the patient received 3 X 2 inhalations of drug or placebo in a cross-over design. The medical staff was blind to the nature of the aerosol (drug or placebo), which was given in random order. The run-in clinical score was high. Asymptomatic days were relatively infrequent and daily drug consumption was high. Functional studies between the cross-over periods showed flow-rate values close to normal, with an increase in residual volume and functional residual capacity. During treatment either with placebo or oxitropium, there was a statistically significant decrease in clinical scores. Results for oxitropium bromide treatment were significantly better than the run-in values (p less than 0.005) and the placebo period (p less than 0.02). There was no significant change in non-trial drug consumption. Functional values showed no difference in terms of flow rate, although oxitropium did cause a significant improvement in the RV/TLC ratio (p less than 0.05). No adverse reactions were reported.