Safety and Efficacy of IL-12 Plasmid DNA Transfection into Pig Skin: Supportive Data for Human Clinical Trials on Gene Therapy and Vaccination.

Gene electrotransfer (GET) of plasmids encoding interleukin 12 (IL-12) has already been used for the treatment of various types of tumors in human oncology and as an adjuvant in DNA vaccines. In recent years, we have developed a plasmid encoding human IL-12 (phIL12) that is currently in a phase I clinical study. The aim was to confirm the results of a non-clinical study in mice on pharmacokinetic characteristics and safety in a porcine model that better resembled human skin. The GET of phIL12 in the skin was performed on nine pigs using different concentrations of plasmid phIL12 and invasive (needle) or noninvasive (plate) types of electrodes. The results of our study demonstrate that the GET of phIL-12 with needle electrodes induced the highest expression of IL-12 at the protein level on day 7 after the procedure. The plasmid was distributed to all tested organs; however, its amount decreased over time and was at a minimum 28 days after GET. Based on plasmid copy number and expression results, together with blood analysis, we showed that IL-12 GET is safe in a porcine animal model. Furthermore, we demonstrated that pigs are a valuable model for human gene therapy safety studies.

A survey study on the recognition and treatment of pain in dogs and cats by Slovenian veterinarians.

OBJECTIVE: To obtain information on pain management in dogs and cats by Slovenian veterinarians, and to compare it with reports from other countries. STUDY DESIGN: Prospective online survey. METHODS: The questions were uploaded using open source survey software and pertained to demographic data, self-evaluation of acquired and current knowledge, attitude towards pain and evaluation of actual knowledge, assessment of chronic and acute pain and pain due to surgery or medical conditions, and the use of analgesics. Descriptive statistics were used to summarize frequency distribution. Wilcoxon signed rank test, Mann-Whitney U test and Spearman's rho were applied where appropriate. RESULTS: The response rate was 11%. From 680 registered members of the Veterinary Chamber of Slovenia, 73 veterinarians completed the survey. Most of the respondents were women (71.2%) working in small (60.3%) or mixed (21.9%) animal practices. A more recent year of graduation was positively correlated to the self-reporting of knowledge acquired during undergraduate studies (rs = 0.339, p = 0.003) and to current knowledge about the topic (rs = 0.293, p = 0.012). Cats were assigned higher scores than dogs for signs of acute (p < 0.001) pain. Female respondents assigned higher scores than male respondents for chronic pain in dogs. Dogs were assigned higher pain scores than cats for surgical procedures and medical conditions. The most frequently used non-steroidal anti-inflammatory drugs (NSAIDs) in dogs and cats were meloxicam and carprofen, and butorphanol was the most commonly used opioid. The choice of NSAID was influenced by perceived effectiveness, experience with use and anti-inflammatory effect. Opioid use was influenced by experience with use and perceived effectiveness. CONCLUSIONS AND CLINICAL RELEVANCE: Slovenian veterinarians acknowledged the importance of pain management as reported for other European countries. However, the use of multimodal analgesia is limited, and inadequacies remain in the pain management of cats.

Pharmacokinetics of carprofen in anaesthetized pigs: a preliminary study.

OBJECTIVE: To investigate the pharmacokinetics of carprofen after a single intravenous (IV) dose and multiple oral doses administered to pigs undergoing electroporation of the pancreas. STUDY DESIGN: Prospective experimental study. ANIMALS: A group of eight female pigs weighing 31.74 ± 2.24 kg (mean ± standard deviation). METHODS: Carprofen 4 mg kg-1 was administered IV after placement of a central venous catheter during general anaesthesia with isoflurane. Blood samples were collected 30 seconds before and 5, 10, 20, 30 and 60 minutes and 2, 4, 6, 8, 12 and 24 hours after carprofen administration. Subsequently, the same dose of carprofen was administered orally, daily, for 6 consecutive days and blood collected at 36, 48, 60, 72, 96, 120, 144 and 168 hours after initial carprofen administration. Plasma was analysed using liquid chromatography with mass spectrometry. Standard pharmacokinetic parameters were calculated by compartmental analysis of plasma concentration-time curves. Data are presented as mean ± standard error. RESULTS: The initial plasma concentration of IV carprofen was estimated at 54.57 ± 3.92 µg mL-1 and decreased to 8.26 ± 1.07 µg mL-1 24 hours later. The plasma elimination curve showed a bi-exponential decline: a rapid distribution phase with a distribution half-life of 0.21 ± 0.03 hours and a slower elimination phase with an elimination half-life of 17.31 ± 3.78 hours. The calculated pharmacokinetic parameters were as follows: the area under the plasma concentration-time curve was 357.3 ± 16.73 µg mL-1 hour, volume of distribution was 0.28 ± 0.07 L kg-1 and plasma clearance rate was 0.19 ± 0.009 mL minute-1 kg-1. The plasma concentration of carprofen, administered orally from days 2 to 7, varied from 9.03 ± 1.87 to 11.49 ± 2.15 µg mL-1. CONCLUSIONS AND CLINICAL RELEVANCE: Carprofen can be regarded as a long-acting non-steroidal anti-inflammatory drug in pigs.

Clinical and histologic tissue responses of skin incisions closed with glycomer 631 and lactomer 9-1 using a subcuticular technique in pigs.

BACKGROUND: Glycomer 631 and lactomer 9-1 are absorbable suture materials indicated for soft tissue approximation in non-infected wounds. Pigs are often used as surgical models in translational research; however, reports of tissue reactions to both suture materials in pigs are limited. The aim of this study was to assess clinical and histologic responses of skin incisions closed with a subcuticular technique using glycomer 631 and lactomer 9-1 in pigs. RESULTS: Skin incisions on 17 pigs were closed with glycomer 631 and lactomer 9-1, and a clinical reactive score (CRS) including erythema, swelling, discharge and dehiscence was calculated on postoperative days 7 and 14. Subcuticular tissue reaction was assessed histologically on postoperative day 14 (the presence of extravascular neutrophils, macrophages, multinucleated giant cells, lymphocytes, fibroblasts, bacterial colonies and the overall severity of the inflammatory response to the suture material), and the cumulative score of the variables was calculated as an aggregate tissue irritation score (ATIS). Tissue samples were examined for suture extrusion and evaluated microbiologically. The clinical reactive score did not differ between the suture materials. Only one ATIS variable, namely the overall severity of the inflammatory response, was lower (p = 0.029) when glycomer 631 was used. Suture extrusion was found in 10/17 of the incisions closed by glycomer 631 and in 7/13 of the incisions closed by lactomer 9-1. Trueperella pyogenes was isolated from the skin and from the area of tissue reaction in six pigs. CONCLUSIONS: No difference in CRS between the suture materials was observed, and thus both materials may be used for the subcuticular technique in pigs. Glycomer 631 induced less tissue reaction only in terms of the overall severity of the inflammatory response. Suture extrusion was observed in more than 50% of incisions regardless of the suture material, possibly due to a large amount of suture material in the wound. Trueperella pyogenes was the only pathogen isolated from the tissue surrounding the suture material.

Antioxidant capacity of lipid- and water-soluble antioxidants in dogs with subclinical myxomatous mitral valve degeneration anaesthetised with propofol or sevoflurane.

BACKGROUND: Antioxidants located in both the hydrophilic and lipophilic compartments of plasma act as a defence system against reactive oxygen species (ROS). Excessive production of ROS during anaesthesia affects the antioxidant capacity of plasma and may result in oxidative stress. The aim of this study was to evaluate the antioxidant capacity of lipid- (ACL) and water-soluble (ACW) antioxidants in client-owned dogs diagnosed with periodontal disease and early-stage myxomatous mitral valve degeneration (MMVD) and anaesthetised for a dental procedure with propofol and sevoflurane or with propofol only. RESULTS: Dogs with MMVD were anaesthetised with propofol and sevoflurane (MMVD/PS, n = 8) or with propofol only (MMVD/P, n = 10). Dogs with no evidence of MMVD (PS, n = 12) were anaesthetised with propofol and sevoflurane. Blood samples for determination of ACL and ACW were collected before and 5 min, 60 min and 6 h after induction to anaesthesia. In MMVD/PS dogs, ACL was significantly higher at all sampling times when compared to PS dogs. Compared to basal values, only anaesthesia maintained with propofol significantly increased ACL at 60 min in dogs with MMVD. In MMVD/P dogs, ACW increased after induction to anaesthesia and remained elevated up to 6 h after anaesthesia. Compared to basal values, anaesthesia maintained with sevoflurane significantly increased ACW only at 60 min in both dogs with and without MMVD. The only difference between propofol and propofol/sevoflurane anaesthesia in dogs with MMVD was significantly higher ACW at 60 min after induction to anaesthesia in the propofol group. CONCLUSIONS: Regarding antioxidant capacity, propofol could be a better choice than sevoflurane for anaesthesia of dogs with early-stage MMVD, although further studies are necessary to clarify the advantage of this antioxidant capacity.

Neurotoxicity of intraneural injection of bupivacaine liposome injectable suspension versus bupivacaine hydrochloride in a porcine model.

OBJECTIVE: To test whether neurotoxic effects of a bupivacaine liposome injectable suspension differ from those of a standard formulation of bupivacaine hydrochloride (HCl) after intraneural injection into the sciatic nerves in pigs. STUDY DESIGN: Prospective, randomized study. ANIMALS: Fifteen pigs, hybrids of Landrace and Large White. METHODS: After the National Ethics Committee approval, 15 pigs were randomly allocated to three groups (n = 5/group) to receive intraneural injections of 4 mL of 1.33% bupivacaine liposome injectable suspension, 0.5% bupivacaine HCl or normal saline. Serial neurologic examinations were conducted to detect sensory and motor response to noxious stimuli using a modified Thalhammer's scale at 2 hour intervals for the first 12 hours after injection and daily thereafter for 2 weeks. Fiber characteristics (density) of the harvested sciatic nerves were measured during histomorphometric analysis. Inflammatory response was studied using immunohistochemical analysis. Data were tested using analyses of variance; p values for paired comparisons were Bonferroni adjusted. RESULTS: Compared with bupivacaine HCl, bupivacaine liposome injectable suspension provided longer sensory (11.2 ± 1.8 hours versus 3.2 ± 1.1 hours, respectively, p < 0.0001) and motor (10.0 ± 2.0 hours versus 4.0 ± 1.4 hours respectively, p < 0.0001) blockade. Histomorphometric parameters were similar among the groups. No changes in axonal density or myelin structure indicative of injury to the sciatic nerves were observed in any of the groups. Number of immunopositive cells did not differ between the bupivacaine liposome injectable suspension (23 ± 6 cells per mm2) and the bupivacaine HCl groups (21 ± 4 cells per mm2), p > 0.90. CONCLUSIONS AND CLINICAL RELEVANCE: Intraneural injections of bupivacaine liposome injectable suspension or bupivacaine HCl in our porcine model did not result in evidence of neurotoxicity.

Cardiac troponin I in dogs anaesthetized with propofol and sevoflurane: the influence of medetomidine premedication and inspired oxygen fraction.

OBJECTIVE: To investigate changes in serum cardiac troponin I (cTnI) concentrations in dogs in which medetomidine was used for sedation or for premedication prior to anaesthesia with propofol and sevoflurane. STUDY DESIGN: Prospective clinical study. ANIMALS: A total of 66 client-owned dogs. METHODS: The dogs were sedated with medetomidine (0.04 mg kg-1) intravenously (IV) (group M; n = 20) and left to breath room air or anaesthetized with propofol (6.5 ± 0.76 mg kg-1 IV) and sevoflurane (4.5% vaporizer setting) in oxygen (group P + S; n = 20) or with medetomidine (0.04 mg kg-1 IV), propofol (1.92 ± 0.63 mg kg-1) and sevoflurane (3% vaporizer setting) in oxygen (group M + P + S; n = 26), respectively. After 35 minutes, medetomidine was antagonized with atipamezole (0.1 mg kg-1 intramuscularly). Blood samples for serum cTnI determination were taken before sedation or anaesthesia, 6 and 12 hours and 4 days thereafter. Serum cTnI concentrations were measured with the Architect STAT Troponin-I assay. RESULTS: Before sedation or anaesthesia, cTnI concentrations were above the detection limit in 22 out of 66 (33%) of dogs. Compared to basal values, cTnI concentrations significantly increased at 6 and 12 hours in all groups and at day 4 in group M. There were no differences in cTnI concentration between groups at baseline, at 6 hours and at 4 days. At 12 hours, cTnI concentrations were significantly higher in groups M and P + S, respectively, compared to group M + P + S. CONCLUSIONS AND CLINICAL RELEVANCE: Oxygenation during anaesthesia and reduction of propofol and sevoflurane dose due to the sparing effects of medetomidine might have played a role in alleviation of myocardial hypoxic injury as indicated by the less severe and short-lived increase of cTnI in the M + P + S group.

Comparison of high definition oscillometric and Doppler ultrasound devices with invasive blood pressure in anaesthetized dogs.

OBJECTIVE: To use the American College of Veterinary Internal Medicine (ACVIM) validation criteria to evaluate the performance of high definition oscillometric (HDO) and Doppler blood pressure measurement techniques against invasive blood pressure measurements in anaesthetized dogs. STUDY DESIGN: Prospective clinical study. ANIMALS: Twenty client-owned dogs. MATERIALS AND METHODS: Invasive blood pressure was measured using a catheter inserted into a pedal artery and an electronic transducer. The sites of cuff placement for the HDO measurements were the mid antebrachium or the proximal tail and, for the Doppler technique, the distal tibia. Agreement between invasive and non-invasive blood pressure measurements was estimated by the Bland-Altman method. RESULTS: Only 10% and 34% of Doppler measurements were within 10 and 20 mmHg of invasive blood pressure values, respectively. The Doppler device failed to meet the ACVIM validation criteria for blood pressure measurement devices. The best agreement between HDO and invasive blood pressure measurement technique was observed for mean arterial blood pressure (MAP); 67% and 95% of readings were within 10 and 20 mmHg of invasive blood pressure values respectively. In addition, 52% and 87% of diastolic arterial blood pressure (DAP) measurements were within 10 and 20 mmHg of invasive readings. High definition oscillometric readings did not meet ACVIM recommended limits for SAP. CONCLUSION AND CLINICAL RELEVANCE: The Doppler technique overestimated and the HDO device showed limited agreement with invasive blood pressure measurement in anaesthetized dogs. High definition oscillometry met most of the ACVIM requirements for MAP and DAP while the Doppler technique did not.

Evaluation of antioxidant status and lipid peroxidation in dogs with myxomatous mitral valve degeneration stage B1.

Myxomatous mitral valve degeneration (MMVD) is the most common naturally occurring heart disease in dogs. There is a lack of data on antioxidant status and oxidative damage in dogs with MMVD stage B1 according to the American College of Veterinary Internal Medicine (ACVIM B1). The aim of this study was to investigate antioxidant status (plasma vitamin E, lipid-standardized vitamin E (LS-VitE), antioxidant capacity of lipid-(ACL) and water-soluble antioxidants, whole blood glutathione peroxidase and erythrocyte superoxide dismutase), and lipid peroxidation [malondialdehyde (MDA)] in dogs with MMVD ACVIM B1. Serum cholesterol and triglyceride concentrations were measured to calculate LS-VitE. Fourteen dogs with MMVD ACVIM B1 and 12 control dogs were included in the study. Dogs with MMVD had significantly higher vitamin E, ACL, MDA, and cholesterol concentrations and significantly higher LS-VitE values than control dogs. No significant correlations between MDA and antioxidant parameters were determined in either group. In conclusion, oxidative damage to lipids is already present and the antioxidant status is altered but not depleted in dogs with MMVD ACVIM B1. The antioxidant response to increased oxidative damage consists mainly of the activation of fat-soluble antioxidants. Further research is needed to evaluate the efficacy and targets of early antioxidant supplementation to prevent or ameliorate oxidative stress and mitigate disease progression in dogs with early-stage MMVD.

Histologic changes of porcine portal vein anastomosis after electrochemotherapy with bleomycin.

Electrochemotherapy (ECT1) is used for treatment of unresectable abdominal malignancies. This study aims to show that ECT of porcine portal vein anastomosis is safe and feasible in order to extend the indications for margin attenuation after resection of locally advanced pancreatic carcinoma. No marked differences were found between the control group and ECT treated groups. Electroporation thus caused irreversible damage to the vascular smooth muscle cells in tunica media that could bedue to the narrow irreversible electroporation zone that may occur near the electrodes, or due to vasa vasorum thrombosis in the tunica externa. Based on the absence of vascular complications, and similar histological changes in lienal veinanastomosis, we can conclude that ECT of portal vein anastomosis is safe and feasible.

Cardiopulmonary Effects and Pharmacokinetics of Dexmedetomidine Used as an Adjunctive Analgesic to Regional Anesthesia of the Oral Cavity with Levobupivacaine in Dogs.

This study investigated the cardiopulmonary effects and pharmacokinetics of dexmedetomidine (DEX) used as an adjunctive analgesic for regional anesthesia of the oral cavity with levobupivacaine in anesthetized dogs. Forty dogs were randomly assigned to four groups of 10 dogs. All dogs received levobupivacaine (4 blocks) with DEX IO (infraorbital block, n = 10) or IA (inferior alveolar block, n = 10) or placebo (PLC; n = 10) or DEX (n = 10) was injected intravenously (IV) after administration of levobupivacaine. The dose of DEX was always 0.5 µg/kg. Cardiopulmonary parameters were recorded, and blood was drawn for the quantification of DEX in plasma using LC-MS/MS. Heart rate was lower in all LB + DEX groups, while mean arterial pressure (MAP) was higher in the LB + DEX IV and LB + DEX IA groups compared to the LB + PLC IV group. Compared to DEX IV, IO and IA administration resulted in lower MAP up to 2 min after application. Absorption of DEX was faster at IO administration (Cmax and Tmax were 0.47 ± 0.08 ng/mL and 7.22 ± 1.28 min and 0.76 ± 0.09 ng/mL and 7.50 ± 1.63 min for the IO and IA block, respectively). The IA administration resulted in better bioavailability and faster elimination (t1/2 was 63.44 ± 24.15 min and 23.78 ± 3.78 min for the IO and IA block, respectively). Perineural administration of DEX may be preferable because of the less pronounced cardiovascular response compared to IV administration.

Transurethral catheterization of female domestic pigs with a Foley catheter.

This report describes transurethral catheterization with a Foley catheter in 19 anaesthetized, laterally recumbent female pigs with a chest circumference of 72.1 ± 4.2 cm and weighing approximately 40 kg. Catheterization within five minutes was successful in 78.9% of the pigs. Catheterization depth, measured as the distance between the bladder neck and the mucocutaneous junction of the vulva, was 14.5 ± 1.3 cm and correlated with chest circumference but not body length. Measurement of catheterization depth aids in optimal positioning and may prevent inflation of the Foley catheter in the urethra.

The pharmacokinetics of levobupivacaine 0.5% after infraorbital or inferior alveolar block in anesthetized dogs.

Introduction: Data are lacking on the pharmacokinetic profile and safety of levobupivacaine (LB) used for regional anesthesia of the maxilla and mandibles in dogs. Methods: Infraorbital block (n = 10), inferior alveolar block (n = 10) or both infraorbital and inferior alveolar blocks (n = 10) were administered to dogs undergoing dental surgery under isoflurane anesthesia. The dose of LB was calculated as 0.11 ml/kg2/3 for the infraorbital block and 0.18 ml/kg2/3 for the inferior alveolar block. Blood samples were collected before and immediately after administration of the oral blocks, and 3, 4, 7, 12, 17, 32, 47, 62, 92, and 122 min thereafter. Quantification of LB in plasma was performed by LC-MS/MS. Results and discussion: The results are presented as median and interquartile range. In dogs in which all four quadrants of the oral cavity were desensitized with LB, the C max was 1,335 (1,030-1,929) ng/ml, the T max was 7 (4-9.5) min, and the AUC(0 → 120) was 57,976 (44,954-96,224) ng min/ml. Plasma concentrations of LB were several times lower than the reported toxic concentrations, and no signs of cardiovascular depression or neurotoxicity were observed in any of the dogs, suggesting that the occurrence of severe adverse effects after administration of LB at the doses used in this study is unlikely.

Numerical mesoscale tissue model of electrochemotherapy in liver based on histological findings.

Electrochemotherapy (ECT) and irreversible electroporation (IRE) are being investigated for treatment of hepatic tumours. The liver is a highly heterogeneous organ, permeated with a network of macro- and microvasculature, biliary tracts and connective tissue. The success of ECT and IRE depends on sufficient electric field established in whole target tissue; therefore, tissue heterogeneity may affect the treatment outcome. In this study, we investigate electroporation in the liver using a numerical mesoscale tissue model. We numerically reconstructed four ECT experiments in healthy porcine liver and computed the electric field distribution using our treatment planning framework. We compared the computed results with histopathological changes identified on microscopic images after treatment. The mean electric field threshold that best fitted the zone of coagulation necrosis was 1225 V/cm, while the mean threshold that best fitted the zone of partially damaged liver parenchyma attributed to IRE was 805 V/cm. We evaluated how the liver macro- and microstructures affect the electric field distribution. Our results show that the liver microstructure does not significantly affect the electric field distribution on the level needed for treatment planning. However, major hepatic vessels and portal spaces significantly affect the electric field distribution, and should be considered when planning treatments.

Slovenian Pet Owners' Experience, Attitudes, and Predictors Regarding Cannabinoid Use in Dogs and Cats.

The aim of this study was to assess the personal experience and attitudes of Slovenian pet owners regarding cannabinoid (CBD) use and to identify the predictors of the first use and reuse of CBDs in dogs and cats. We hypothesized that positive attitudes toward CBDs, postmodern health values, and personal experience would be significant predictors of CBD use in animals. An open online survey targeted randomly selected Slovenian dog and cat owners, regardless of their experience with cannabis products. The questionnaire consisted of six sections related to demographic data and personal experience with CBD use, information about the participant's animal, experience with CBD use in the participant's animal, reasons for not using CBDs in their animal, attitudes toward CBD use in dogs and cats, and postmodern health values. Descriptive statistics were performed to analyze demographics, personal experience with CBD use, and experience with CBD use in dogs and cats. Hierarchical multiple regression using the enter method was performed to analyze the important predictors of CBD use. A total of 408 completed questionnaires were included in the statistical analysis. A substantial proportion (38.5%) of owners had already used CBDs to treat their animal. Positive attitudes and previous personal experience were significant (p < 0.05) predictors of first use and reuse of CBDs in pets, while postmodern health values were not. In conclusion, the decision to use CBDs for medicinal purposes is based on acquired information and personal experience. Veterinarians should be informed and familiar with CBDs as a treatment option. However, further research is essential to establish the use of CBDs in veterinary medicine. Improved laws and regulations are also needed to ensure that only high-quality medications are prescribed to dogs and cats.

Rescue oxygenation success by cannula or scalpel-bougie emergency front-of-neck access in an anaesthetised porcine model.

In the obese, the evidence for the choice of the optimal emergency front-of-neck access technique is very limited and conflicting. We compared cannula and scalpel-bougie emergency front-of-neck access techniques in an anaesthetised porcine model with thick pretracheal tissue. Cannula and scalpel-bougie cricothyroidotomy techniques were performed in 11 and 12 anaesthetised pigs, respectively. Following successful tracheal access, oxygenation was commenced and continued for 5 min using Rapid-O2 device for cannula and circle breathing system for scalpel-bougie study groups. The primary outcome was a successful rescue oxygenation determined by maintenance of arterial oxygen saturation >90% 5 min after the beginning of oxygenation. Secondary outcomes included success rate of airway device placement, time to successful airway device placement, and trauma to the neck and airway. The success rate of rescue oxygenation was 18% after cannula, and 83% after scalpel-bougie technique (P = 0.003). The success rate of airway device placement was 73% with cannula and 92% with scalpel-bougie technique (P = 0.317). Median (inter-quartile-range) times to successful airway device placement were 108 (30-256) and 90 (63-188) seconds (P = 0.762) for cannula and scalpel-bougie emergency front-of-neck access, respectively. Proportion of animals with iatrogenic trauma additional to the procedure itself was 27% for cannula and 75% for scalpel-bougie technique (P = 0.039). Thus, in the porcine model of obesity, the scalpel-bougie technique was more successful in establishing and maintaining rescue oxygenation than cannula-based technique; however, it was associated with a higher risk of severe trauma.

Case Report: Intoxication in a Pig (Sus Scrofa Domesticus) After Transdermal Fentanyl Patch Ingestion.

An experimental study on the effects of electroporation on pancreatic tissue was performed in pigs, and the fentanyl transdermal patch (FTP) was used postoperatively as part of multimodal pain management. Ingestion of an FTP, which resulted in fentanyl intoxication, was suspected 5 days after placement in one of the experimental pigs. The pig was first dysphoric, running in the stall, panting and vocalizing until it finally became depressed and it remained lying on the floor. Ingestion of an FTP was not observed but the fentanyl plasma concentration on the day of intoxication was 20.7 ng/ml, while at its peak after FTP administration it was only 0.492 ng/ml. The intoxication was successfully treated with a single intramuscular naloxone injection.

Safety and Feasibility of Electrochemotherapy of the Pancreas in a Porcine Model.

OBJECTIVES: The use of thermal ablative therapies in the pancreatic tumors is limited because of the risk of the vessel injury and potential pancreatitis or fistula formation. Electrochemotherapy (ECT) is an ablative therapy with an established role in the treatment of cutaneous and liver tumors. This study was designed to evaluate the safety and feasibility of ECT of the pancreas in a porcine survival model. METHODS: In the first group, 4 animals underwent computed tomography (CT)-guided percutaneous ECT with bleomycin of the pancreatic tail. In the second group (4 animals), the intraoperative ECT with bleomycin of pancreatic tail and head was performed. Animals were followed for 7 days and then killed. Clinical parameters, CT imaging, laboratory, and histologic analysis were performed. RESULTS: All pigs survived the ECT procedure and none of them developed clinical signs of acute pancreatitis or related complications. There were no signs of acute pancreatitis or damage to the large vessels present in the follow-up CT scans. No significant change in laboratory parameters was obtained after procedure. CONCLUSIONS: This study shows that ECT with bleomycin is feasible and safe in the pancreatic parenchyma. Clinical studies are needed to evaluate the efficacy of ECT in pancreatic cancer.

Radiological findings of porcine liver after electrochemotherapy with bleomycin.

Background Radiologic findings after electrochemotherapy of large hepatic blood vessels and healthy hepatic parenchyma have not yet been described. Materials and methods We performed a prospective animal model study with regulatory approval, including nine grower pigs. In each animal, four ultrasound-guided electroporated regions were created; in three regions, electrodes were inserted into the lumen of large hepatic vessels. Two types of electrodes were tested; variable linear- and fixed hexagonal-geometry electrodes. Ultrasonographic examinations were performed immediately and up to 20 minutes after the procedure. Dynamic computed tomography was performed before and at 60 to 90 minutes and one week after the procedure. Results Radiologic examinations of the treated areas showed intact vessel walls and patency; no hemorrhage or thrombi were noted. Ultrasonographic findings were dynamic and evolved from hyperechogenic microbubbles along electrode tracks to hypoechogenicity of treated parenchyma, diffusion of hyperechogenic microbubbles, and hypoechogenicity fading. Contrast-enhanced ultrasound showed decreased perfusion of the treated area. Dynamic computed tomography at 60 to 90 minutes after the procedure showed hypoenhancing areas. The total hypoenhancing area was smaller after treatment with fixed hexagonal electrodes than after treatment with variable linear geometry electrodes. Conclusions Radiologic findings of porcine liver after electrochemotherapy with bleomycin did not show clinically significant damage to the liver, even if a hazardous treatment strategy, such as large vessel intraluminal electrode insertion, was employed, and thus further support safety and clinical use of electrochemotherapy for treatment of hepatic neoplasia.

Large Liver Blood Vessels and Bile Ducts Are Not Damaged by Electrochemotherapy with Bleomycin in Pigs.

The first clinical studies on the use of electrochemotherapy to treat liver tumours that were not amenable to surgery or thermal ablation techniques have recently been published. However, there is still a lack of data on the effects of electrochemotherapy on normal liver tissue. Therefore, we designed a translational animal model study to test whether electrochemotherapy with bleomycin causes clinically significant damage to normal liver tissue, with emphasis on large blood vessels and bile ducts. We performed electrochemotherapy with bleomycin or delivered electric pulses alone using a potentially risky treatment strategy in eight pigs. Two and seven days after treatment, livers were explanted, and histological analysis was performed. Blood samples were collected before treatment and again before euthanasia to evaluate blood biomarkers of liver function and systemic inflammatory response. We found no thrombosis or other clinically significant damage to large blood vessels and bile ducts in the liver. No clinical or laboratory findings suggested impaired liver function or systemic inflammatory response. Electrochemotherapy with bleomycin does not cause clinically significant damage to normal liver tissue. Our study provides further evidence that electrochemotherapy with bleomycin is safe for treatment of patients with tumours near large blood vessels in the liver.