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Delivery of adeno-associated viral vectors (AAVs) to cerebrospinal fluid (CSF) has emerged as a promising approach to achieve widespread transduction of the central nervous system (CNS) and peripheral nervous system (PNS), with direct applicability to the treatment of a wide range of neurological diseases, particularly lysosomal storage diseases. Although studies in small animal models have provided proof of concept and experiments in large animals demonstrated feasibility in bigger brains, there is not much information on long-term safety or durability of the effect. Here, we report a 7-year study in healthy beagle dogs after intra-CSF delivery of a single, clinically relevant dose (2 × 1013 vg/dog) of AAV9 vectors carrying the canine sulfamidase, the enzyme deficient in mucopolysaccharidosis type IIIA. Periodic monitoring of CSF and blood, clinical and neurological evaluations, and magnetic resonance and ultrasound imaging of target organs demonstrated no toxicity related to treatment. AAV9-mediated gene transfer resulted in detection of sulfamidase activity in CSF throughout the study. Analysis at tissue level showed widespread sulfamidase expression and activity in the absence of histological findings in any region of encephalon, spinal cord, or dorsal root ganglia. Altogether, these results provide proof of durability of expression and long-term safety for intra-CSF delivery of AAV-based gene transfer vectors encoding therapeutic proteins to the CNS., Graphical abstract, Seven-year follow-up of dogs after intra-CSF administration of AAV9-sulfamidase vectors results in detection of sulfamidase activity in CSF and widespread transgene expression in CNS, PNS, and liver, in the absence of any adverse events. Proof of durability and safety of this gene therapy support its clinical translation to treat CNS diseases.

Objective To determine the cis-atracurium pharmacokinetic data and laudanosine production of a single 1 mg kg–1 cis-atracurium dose in the pig and to compare the pharmacokinetics between two groups of different ages. Study design Prospective experimental study. Animals Sixteen female pigs in two groups. Group A included eight animals aged 2.0–2.5 months and weighed 26.6 ± 3.6 kg. Group B included eight animals aged 4.0–5.0 months and weighed 57.4 ± 8.3 kg. Methods The pigs were anaesthetized and monitored throughout the procedure. Arterial blood samples collected at 0, 0.5, 1, 2, 5, 10, 20, 30, 45, 60, 90, 120 and 180 minutes after cis-atracurium injection were cooled and centrifuged. Plasma was acidified and stored at –20 °C for subsequent cis-atracurium and laudanosine analyses. Results Anaesthetic parameters were within normal ranges throughout the procedure. Plasma cis-atracurium and laudanosine concentrations were measured for the 16 pigs. Elimination rate constant, elimination half-life, area under the curve, mean residence time, distribution volume and total clearance were calculated for each pig. Statistical differences (p Conclusion and clinical relevance Longer duration of plasma cis-atracurium concentrations were observed in younger pigs. Distribution volume was also higher in younger pigs. Conversely, total clearance and area under the curve were similar between the two age groups. No laudanosine production was detected, suggesting a different cis-atracurium metabolism in the pig compared with other species.

[EN] Radiofrequency energy has been used both experimentally and clinically to manage the pancreatic remnant after distal pancreatectomies. Our goal was to determine whether endoluminal radiofrequency (RF) ablation of the main pancreatic duct in large animals would be more efficient than glue occlusion as an exocrine pancreatic atrophy-inducing procedure. Thirty-four Landrace pigs were assigned to either the transpapilar (n = 16) or transection (n = 18) groups. The transection implied the pancreas neck was severed. In each of these groups the remaining distal pancreatic duct was occluded either by RF or by glue. In the transpapilar group complete atrophy was observed in all the RF cases, while atrophy was incomplete in all the members of the glue subgroup. The failure rate of the main pancreatic duct (usually expressed by a pseudocyst) in the transection groups was dramatically higher in the glue subgroup than the RF subgroups (9 out of 9 and 1 out of 9, respectively) and postoperative mortality occurred only in the glue subgroup (3 out of 9). These results show the superiority of endoluminal RF ablation over glue for main pancreatic duct occlusion, as seen by the degree of atrophy and fewer postoperative pancreatic fistulas., This work was supported by the Spanish Ministerio de Economia, Industria y Competitividad under the "Plan Estatal de Investigacion, Desarrollo e Innovacion Orientada a los Retos de la Sociedad" Grant TEC2014-52383-C3-R (TEC2014-52383-C3-1-R and TEC2014-52383-C3-3-R). Elzbieta Ewertowska has a Predoctoral Grant (BES-2015-073285) from the Ministry of Economy, Industry and Competitiveness (Government of Spain).

[EN] Purpose Multiple attempts have been made to manage the pancreatic stump and the pancreatic duct in order to reduce the rate of postoperative pancreatic fistula (POPF) after pancreaticoduodenectomy (PD), however radiofrequency-based technologies could help to achieve this goal. Previous encouraging clinical and experimental results support the use of endoluminal thermal ablation (ETHA) of the main pancreatic duct to reduce pancreatic exocrine secretion and hence POPF. We here describe our initial clinical experience with ETHA of the main pancreatic duct in two cases at high risk of POPF. Methods Two cases underwent PD for malignancy with a high risk of POPF (adenocarcinoma, obese patients, surgical difficulties with heavy intraoperative blood loss, soft pancreas or walled-off pancreatitis and a tight small pancreatic main duct). In both cases, ETHA of the main pancreatic duct was conducted intraoperatively just before Blumgart-type pancreatic-jejunal anastomosis using a ClosureFast catheter (Medtronic, Mansfield, MA, USA) normally used for varicose vein treatment (therefore an off-label use). Results Although a clear radiological POPF was detected in the second case, the clinical postoperative course in both cases was uneventful. Little pancreatic fluid collected in the abdominal drainage with low levels of amylase enzyme, confirming low exocrine pancreatic function. No other procedure-related complications were detected. Conclusion Endoluminal thermal ablation of the main pancreatic duct may be a feasible and safe technique to reduce the adverse effects of POPF after PD., This work was supported by the Spanish Ministerio de Ciencia, Innovacion y Universidades under "Programa Estatal de IthornDthorni Orientada a los Retos de la Sociedad", Grants RTI2018-094357-B-C21 and RTI2018-094357-B-C22.

The "One world, one health" initiative emphasizes the need for new strategies to control human and animal tuberculosis (TB) based on their shared interface. A good example would be the development of novel universal vaccines against Mycobacterium tuberculosis complex (MTBC) infection. This study uses the goat model, a natural TB host, to assess the protective effectiveness of a new vaccine candidate in combination with Bacillus Calmette-Guerin (BCG) vaccine. Thirtythree goat kids were divided in three groups: Group 1) vaccinated with BCG (week 0), Group 2) vaccinated with BCG and boosted 8 weeks later with a recombinant adenovirus expressing the MTBC antigens Ag85A, TB10.4, TB9.8 and Acr2 (AdTBF), and Group 3) unvaccinated controls. Later on, an endobronchial challenge with a low dose of M. caprae was performed (week 15). After necropsy (week 28), the pulmonary gross pathology was quantified using high resolution Computed Tomography. Small granulomatous pulmonary lesions (< 0.5 cm diameter) were also evaluated through a comprehensive qualitative histopathological analysis. M. caprae CFU were counted from pulmonary lymph nodes. The AdTBF improved the effects of BCG reducing gross lesion volume and bacterial load, as well as increasing weight gain. The number of Ag85A-specific gamma interferon-producing memory T-cells was identified as a predictor of vaccine efficacy. Specific cellular and humoral responses were measured throughout the 13-week post-challenge period, and correlated with the severity of lesions. Unvaccinated goats exhibited the typical pathological features of active TB in humans and domestic ruminants, while vaccinated goats showed only very small lesions. The data presented in this study indicate that multi-antigenic adenoviral vectored vaccines boosts protection conferred by vaccination with BCG. © 2013 Pérez de Val et al.

The aim of the study was to determine the distribution of different volumes of methylene blue solution injected into the epidural space in anaesthetized pregnant and non-pregnant sheep, to evaluate its cranial distribution and to compare between them. Fifteen pregnant and fifteen non-pregnant sheep were included in the study. Sheep were anaesthetized and received 0.05, 0.1, or 0.2 mL/kg of a lumbosacral epidural solution containing 0.12% methylene blue in 0.9% saline. Thirty minutes after the epidural injection, the ewes were euthanized. The extension of the dye within the epidural space was measured, and the correlation between the volume of the dye injected and the number of stained vertebrae was evaluated. The cranial migration of the dye between pregnant and non-pregnant sheep was also compared. The results show that the volume of methylene blue injected epidurally into pregnant and non-pregnant sheep correlated directly with its cephalic distribution into the epidural space; and a volume of 0.1 mL/kg or 0.2 mL/kg stained up to the first lumbar segment in pregnant and non-pregnant sheep, respectively. Also, the results suggest that the volume of drugs administered into the epidural space of pregnant sheep should be half the volume that would be used in non-pregnant sheep.

[EN] Purpose: To improve the computer modelling of radiofrequency ablation (RFA) by internally cooled wet (ICW) electrodes with added clinically oriented features. Methods: An improved RFA computer model by ICW electrode included: (1) a realistic spatial distribution of the infused saline, and (2) different domains to distinguish between healthy tissue, saline-infused tumour, and non-infused tumour, under the assumption that infused saline is retained within the tumour boundary. A realistic saline spatial distribution was obtained from an in vivo pig liver study. The computer results were analysed in terms of impedance evolution and coagulation zone (CZ) size, and were compared to the results of clinical trials conducted on 17 patients with the same ICW electrode. Results: The new features added to the model provided computer results that matched well with the clinical results. No roll-offs occurred during the 4-min ablation. CZ transversal diameter (4.10 +/- 0.19 cm) was similar to the computed diameter (4.16 cm). Including the tumour and saline infusion in the model involved (1) a reduction of the initial impedance by 10 - 20 ohm, (2) a delay in roll-off of 20 s and 70 - 100 s, respectively, and (3) 18 - 31% and 22 - 36% larger CZ size, respectively. The saline spatial distribution geometry was also seen to affect roll-off delay and CZ size. Conclusions: Using a three-compartment model and a realistic saline spatial distribution notably improves the match with the outcome of the clinical trials., This work was supported by the Spanish Ministerio de Economia, Industria y Competitividad under the "Plan Estatal de Investigacion, Desarrollo e Innovacion Orientada a los Retos de la Sociedad" Grant TEC2014-52383-C3-R (TEC2014-52383-C3-1-R and TEC2014-52383-C3-3-R). Elzbieta Ewertowska has a Predoctoral Grant (BES-2015-073285) from the Ministry of Economy, Industry and Competitiveness (Government of Spain).

Neuromuscular blocking agents should be included as part of a balanced anaesthetic protocol to improve anaesthetic management, although doses are not always established for each species. Cis-atracurium is a benzylisoquinolinium neuromuscular blocking agent with an intermediate duration of action devoid of significant adverse effects previously used in pigs with a wide dosage range. Cis-atracurium was administered at 1 mg/kg bolus to sixteen pigs to establish its time profile and effects. The pigs were premedicated intramuscularly with 4 mg/kg azaperone, 8 mg/kg ketamine and 0.2 mg/kg morphine IM and maintained with isoflurane in oxygen. After cis-atracurium administration, neuromuscular monitoring via acceleromyography was started until the recovery of the 90% of the train of four ratio. Complete decrease in the train of four ratio was accomplished in eleven pigs. Onset of action was 70 s, with a recovery of the fourth twitch at 26 min and a recovery of a train of four ratio greater than 90% in 60 min. In conclusion, 1 mg/kg intravenous cis-atracurium in the pig allowed for a rapid onset of action and a complete recovery after 60 min although high variability in the time profile is seen.

To assess the validity and reliability of Doppler ultrasonography (DOP) as compared with invasive arterial blood pressure measurements in anaesthetized dogs weighing less than 5 kg.Controlled, prospective, clinical study.A total of 41 privately owned dogs weighing less than 5 kg.The dogs were anaesthetized, and an intra-arterial catheter was placed aseptically in the dorsal pedal artery of the pelvic limb to perform invasive blood pressure (IBP) measurement. The contralateral metatarsal surface of the foot was clipped in order to perform DOP. Both techniques were used to record blood pressure measurements every 5 minutes during surgical procedures. The blood pressure measurements were categorized into two groups: hypotensive [mean arterial pressure (MAP)60 mmHg] and normotensive (MAP between 60 and 120 mmHg). A linear mixed model was used to compare the DOP and IBP values. The results were evaluated according to the requirements of the American College of Veterinary Internal Medicine (ACVIM) guidelines for the validation of devices.DOP provided higher values compared to the systolic arterial blood pressure (SAP) and MAP of IBP measurements. The closest agreement between the two techniques was found for SAP; the bias was 8.8, and limits of agreement (LOA) were -32.9 and 50.4. Similar results were observed when the IBP technique was categorized. The closest agreement was for SAP in animals categorized as normotensive; the bias was 8.2, and LOAs were -32.8 and 49.2. The level of agreement between DOP and IBP did not meet the ACVIM recommendations.Our results suggest there is poor agreement between DOP and IBP measurements in anaesthetized dogs weighing less than 5 kg. Hence, the use of DOP in these animals could be misleading.

[EN] Introduction: Endoluminal sealing of the pancreatic duct by glue or sutures facilitates the management of the pancreatic stump. Our objective was to develop a catheter-based alternative for endoluminal radiofrequency (RF) sealing of the pancreatic duct. Materials and methods: We devised a novel RF ablation technique based on impedance-guided catheter pullback. First, bench tests were performed on ex vivo models to tune up the technique before the in vivo study, after which endoluminal RF sealing of a similar to 10 cm non-transected pancreatic duct was conducted on porcine models using a 3 Fr catheter. After 30 days, sealing effectiveness was assessed by a permeability test and a histological analysis. Results: The RF technique was feasible in all cases and delivered similar to 5 W of power on an initial impedance of 308 +/- 60 ohm. Electrical impedance evolution was similar in all cases and provided guidance for modulating the pullback speed to avoid tissue sticking and achieve a continuous lesion. During the follow-up the animals rate of weight gain was significantly reduced (p < 0.05). Apart from signs of exocrine atrophy, no other postoperative complications were found. At necropsy, the permeability test failed and the catheter could not be reintroduced endoluminally, confirming that sealing had been successful. The histological analysis revealed a homogeneous exocrine atrophy along the ablated segment in all the animals. Conclusions: Catheter-based RF ablation could be used effectively and safely for endoluminal sealing of the pancreatic duct. The findings suggest that a fully continuous lesion may not be required to obtain complete exocrine atrophy., This work was supported by the Spanish Ministerio de Economia, Industria y Competitividad under "Plan Estatal de Investigacion, Desarrollo e Innovacion Orientada a los Retos de la Sociedad" (Grant TEC2014-52383-C3-R) and by the Spanish Ministerio de Ciencia, Innovacion y Universidades under "Programa Estatal de I+D+i Orientada a los Retos de la Sociedad" (Grant: RTI2018-094357-B-C21 and -C22). Elzbieta Ewertowska has a Predoctoral Grant (BES-2015-073285) from the Ministry of Economy, Industry and Competitiveness (Government of Spain). Also funding was received from Secretaria de Estado de Investigacion, Desarrollo e Innovacion.

Objective To investigate the diagnostic and prognostic value over time of plasma iron compared with the inflammatory markers albumin, C-reactive protein (CRP), and fibrinogen in dogs with systemic inflammatory response syndrome (SIRS). Design Prospective observational study of sequentially enrolled dogs. Setting ICU of a veterinary teaching hospital. Animals One hundred and sixteen client-owned dogs: 54 dogs with SIRS or sepsis, 42 with focal inflammation, and 20 clinically healthy dogs. Measurements and Main Results Blood samples were obtained on admission in all study groups, and then on alternate days until discharge or death in both inflammation groups. On admission, dogs with SIRS had significantly lower plasma iron (65 ± 5.8 μg/dL, P = 0.001) concentrations than dogs with focal inflammation (89.5 ± 6.2 μg/dL, P = 0.001). Plasma iron, albumin, and CRP effectively discriminated the SIRS/sepsis group from those presenting with focal inflammation with areas under the curve for the receiver operating curves of 0.679, 0.834, and 0.704, respectively. The admission values for these variables did not discriminate survivors from nonsurvivors within the SIRS/sepsis group. However, the magnitude of increase in iron concentration and the decrease in CRP concentration from admission to hospital discharge was higher in survivors than in nonsurvivors within the SIRS/septic group (22.8 vs. 2.51 μg/dL, respectively, P = 0.021 for iron; −67.1 vs. −4.1 mg/L, respectively, P = 0.002 for CRP), resulting in iron and CRP concentrations at hospital discharge for survivors similar to those in the focal inflammation group. Conclusion Hypoferremia is a sensitive marker of systemic inflammation in dogs. In this study, the increase in iron concentrations during the hospitalization period of SIRS/septic dogs was associated with a better prognosis, suggesting that plasma iron in combination with CRP and albumin concentrations might be used to monitor dogs with inflammatory disease processes.

To determine the cis-atracurium pharmacokinetic data and laudanosine production of a single 1 mg kgProspective experimental study.Sixteen female pigs in two groups. Group A included eight animals aged 2.0-2.5 months and weighed 26.6 ± 3.6 kg. Group B included eight animals aged 4.0-5.0 months and weighed 57.4 ± 8.3 kg.The pigs were anaesthetized and monitored throughout the procedure. Arterial blood samples collected at 0, 0.5, 1, 2, 5, 10, 20, 30, 45, 60, 90, 120 and 180 minutes after cis-atracurium injection were cooled and centrifuged. Plasma was acidified and stored at -20 °C for subsequent cis-atracurium and laudanosine analyses.Anaesthetic parameters were within normal ranges throughout the procedure. Plasma cis-atracurium and laudanosine concentrations were measured for the 16 pigs. Elimination rate constant, elimination half-life, area under the curve, mean residence time, distribution volume and total clearance were calculated for each pig. Statistical differences (p0.05) in the elimination rate constant, elimination half-life, mean residence time and distribution volume values were observed between the two groups. Elimination half-life, mean residence time and distribution volume values were higher and elimination rate constant lower in younger pigs than in older pigs. No plasma laudanosine concentrations were detected in any pig.Longer duration of plasma cis-atracurium concentrations were observed in younger pigs. Distribution volume was also higher in younger pigs. Conversely, total clearance and area under the curve were similar between the two age groups. No laudanosine production was detected, suggesting a different cis-atracurium metabolism in the pig compared with other species.

[Background]: Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder characterized by progressive muscle weakness, paralysis and death. There is no effective treatment for ALS and stem cell therapy has arisen as a potential therapeutic approach., [Methods]: SOD1 mutant mice were used to study the potential neurotrophic effect of bone marrow cells grafted into quadriceps femoris muscle., [Results]: Bone marrow intramuscular transplants resulted in increased longevity with improved motor function and decreased motoneuron degeneration in the spinal cord. Moreover, the increment of the glial-derived neurotrophic factor and neurotrophin 4 observed in the grafted muscles suggests that this partial neuroprotective effect is mediated by neurotrophic factor release at the neuromuscular junction level. Finally, certain neurodegeneration and muscle disease-specific markers, which are altered in the SOD1G93A mutant mouse and may serve as molecular biomarkers for the early detection of ALS in patients, have been studied with encouraging results., [Conclusions]: This work demonstrates that stem cell transplantation in the muscle prolonged the lifespan, increased motoneuron survival and slowed disease progression, which was also assessed by genetic expression analysis., This study was supported by the Spanish State Research Agency “Severo Ochoa” Program for Centers of Excellence in R&D (SEV-2013-0317) and Instituto de Salud Carlos III. Red de Terapia Celular, TERCEL (RD12/0019/0024). Project “RD16/001/0010”, funded by Instituto de Salud Carlos III and co-funded by European Union (ERDF/ESF, “Investing in your future”)

A finite element model of radiofrequency tumor ablation with an internally cooled wet electrode was built and its computer results were compared to those reported in a clinical study involving 17 patients with hepatocellular carcinoma. The model whose results best matched the clinical results was based on three compartments and assumed a hydrated tumor: computed transversal diameter of 39.2 mm vs. reported diameter of 33.7 ± 7.1 mm. The impedances of the model (47.9 Ω) and the clinical trials (46 ± 6 Ω) were also similar, both at 140 W. The results showed the advantage of a three-compartment over a two-compartment model when simulating the real effects of treatment on cancerous tissue.

Irreversible electroporation (IRE) has recently gained in popularity as an ablative technique, however little is known about its oncological long-term outcomes. To determine the long-time survival of animals treated with a high dose of IRE and which histological changes it induces in tumoral tissue, IRE ablation was performed in forty-six athymic-nude mice with KM12C tumors implanted in the liver by applying electric current with different voltages (2000 V/cm, 1000 V/cm). The tumors were allowed to continue to grow until the animals reached the end-point criteria. Histology was harvested and the extent of tumor necrosis was semi-quantitatively assessed. IRE treatment with the 2000 V/cm protocol significantly prolonged median mouse survival from 74.3 ± 6.9 days in the sham group to 112.5 ± 15.2 days in the 2000 V/cm group. No differences were observed between the mean survival of the 1000 V/cm and the sham group (83.2 ± 16.4 days, p = 0.62). Histology revealed 63.05% ± 23.12 of tumor necrosis in animals of the 2000 V/cm group as compared to 17.50% ± 2.50 in the 1000 V/cm group and 25.6% ± 22.1 in the Sham group (p = 0.001). IRE prolonged the survival of animals treated with the highest electric field (2000 V/cm). The animals in this group showed significantly higher rate of tumoral necrosis. This research was supported by the spanish government under grants TEC2011-27133-c02 and TEC2010-17285, and by the spanish “plan estatal de investigación, desarrollo e innovación orientada a los retos de la sociedad” under grant TEC2014–52383-c3-r (TEC2014–52383-c3-2-r and TEC2014–52383-c3-3-r).

Metastases in the liver frequently grow as scattered tumor nodules that neither can be removed by surgical resection nor focally ablated. Previously, we have proposed a novel technique based on irreversible electroporation that may be able to simultaneously treat all nodules in the liver while sparing healthy tissue. The proposed technique requires increasing the electrical conductivity of healthy liver by injecting a hypersaline solution through the portal vein. Aiming to assess the capability of increasing the global conductivity of the liver by means of hypersaline fluids, here, it is presented a mathematical model that estimates the NaCl distribution within the liver and the resulting conductivity change. The model fuses well-established compartmental pharmacokinetic models of the organ with saline injection models used for resuscitation treatments, and it considers changes in sinusoidal blood viscosity because of the hypertonicity of the solution. Here, it is also described a pilot experimental study in pigs in which different volumes of NaCl 20% (from 100 to 200 mL) were injected through the portal vein at different flow rates (from 53 to 171 mL/minute). The in vivo conductivity results fit those obtained by the model, both quantitatively and qualitatively, being able to predict the maximum conductivity with a 14.6% average relative error. The maximum conductivity value was 0.44 second/m, which corresponds to increasing 4 times the mean basal conductivity (0.11 second/m). The results suggest that the presented model is well suited for predicting on liver conductivity changes during hypertonic saline injection.

For most lysosomal storage diseases (LSDs) affecting the CNS, there is currently no cure. The BBB, which limits the bioavailability of drugs administered systemically, and the short half-life of lysosomal enzymes, hamper the development of effective therapies. Mucopolysaccharidosis type IIIA (MPS IIIA) is an autosomic recessive LSD caused by a deficiency in sulfamidase, a sulfatase involved in the stepwise degradation of glycosaminoglycan (GAG) heparan sulfate. Here, we demonstrate that intracerebrospinal fluid (intra-CSF) administration of serotype 9 adenoassociated viral vectors (AAV9s) encoding sulfamidase corrects both CNS and somatic pathology in MPS IIIA mice. Following vector administration, enzymatic activity increased throughout the brain and in serum, leading to whole body correction of GAG accumulation and lysosomal pathology, normalization of behavioral deficits, and prolonged survival. To test this strategy in a larger animal, we treated beagle dogs using intracisternal or intracerebroventricular delivery. Administration of sulfamidase-encoding AAV9 resulted in transgenic expression throughout the CNS and liver and increased sulfamidase activity in CSF. High-titer serum antibodies against AAV9 only partially blocked CSF-mediated gene transfer to the brains of dogs. Consistently, anti-AAV antibody titers were lower in CSF than in serum collected from healthy and MPS IIIA-affected children. These results support the clinical translation of this approach for the treatment of MPS IIIA and other LSDs with CNS involvement.

Diabetes is associated with severe secondary complications, largely caused by poor glycemic control. Treatment with exogenous insulin fails to prevent these complications completely, leading to significant morbidity and mortality. We previously demonstrated that it is possible to generate a “glucose sensor” in skeletal muscle through coexpression of glucokinase and insulin, increasing glucose uptake and correcting hyperglycemia in diabetic mice. Here, we demonstrate long-term efficacy of this approach in a large animal model of diabetes. A one-time intramuscular administration of adeno-associated viral vectors of serotype 1 encoding for glucokinase and insulin in diabetic dogs resulted in normalization of fasting glycemia, accelerated disposal of glucose after oral challenge, and no episodes of hypoglycemia during exercise for >4 years after gene transfer. This was associated with recovery of body weight, reduced glycosylated plasma proteins levels, and long-term survival without secondary complications. Conversely, exogenous insulin or gene transfer for insulin or glucokinase alone failed to achieve complete correction of diabetes, indicating that the synergistic action of insulin and glucokinase is needed for full therapeutic effect. This study provides the first proof-of-concept in a large animal model for a gene transfer approach to treat diabetes.

A six-month-old male entire golden retriever was presented with an umbilical mass noted by the owners four days before. Clinical examination revealed fever (39.7°C) and an umbilical fistulised abscess. Abdominal palpation revealed a solid mass in the caudal abdomen. The ultrasonographic study confirmed the presence of an abdominal mass (7 cm in diameter) adhered to several abdominal organs, mild localised peritonitis and an umbilical defect. The mass was compatible with an abscess or a granuloma potentially associated with an urachal remnant. An exploratory laparotomy confirmed the suspicion. The mass was removed and the urachal remnant corrected. Histopathology revealed the presence of a granuloma with multiple necrotic areas and basophilic structures suggestive of bacterial infection. Few days after surgery, all clinical signs resolved completely. Urachal remnants are rarely reported in small animals. To the authors’ knowledge, this is the first report describing such a severe presentation with complete resolution.

Background Spread hepatic tumours are not suitable for treatment either by surgery or conventional ablation methods. The aim of this study was to evaluate feasibility and safety of selectively increasing the healthy hepatic conductivity by the hypersaline infusion (HI) through the portal vein. We hypothesize this will allow simultaneous safe treatment of all nodules by irreversible electroporation (IRE) when applied in a transhepatic fashion. Material and methods Sprague Dawley (Group A, n = 10) and Athymic rats with implanted hepatic tumour (Group B, n = 8) were employed. HI was performed (NaCl 20%, 3.8 mL/Kg) by trans-splenic puncture. Deionized serum (40 mL/Kg) and furosemide (2 mL/Kg) were simultaneously infused through the jugular vein to compensate hypernatremia. Changes in conductivity were monitored in the hepatic and tumour tissue. The period in which hepatic conductivity was higher than tumour conductivity was defined as the therapeutic window (TW). Animals were monitored during 1-month follow-up. The animals were sacrificed and selective samples were used for histological analysis. Results The overall survival rate was 82.4% after the HI protocol. The mean maximum hepatic conductivity after HI was 2.7 and 3.5 times higher than the baseline value, in group A and B, respectively. The mean maximum hepatic conductivity after HI was 1.4 times higher than tumour tissue in group B creating a TW to implement selective IRE. Conclusions HI through the portal vein is safe when the hypersaline overload is compensated with deionized serum and it may provide a TW for focused IRE treatment on tumour nodules.

Purpose: To evaluate the impact of using monopolar thermal coagulation based on radiofrequency (RF) currents on intraoperative blood loss during liver resection. Materials and methods: A prospective randomized controlled trial was planned. Patients undergoing hepatectomy were randomized into two groups. In the Control group (n=10), hemostasis was obtained with a combination of stitches, vessel-sealing bipolar RF systems, sutures or clips. In the monopolar radiofrequency coagulation (MRFC) group (n=18), hemostasis was mainly obtained using an internally cooled monopolar RF electrode. Results: No differences in demographic or clinical characteristics were found between groups. Mean blood loss during liver resection in the Control group was more than twice that of the MRFC group (556±471 ml vs. 225±313 ml, p=0.02). The adjusted mean bleeding/transection area was also significantly higher in the Control group (7.0±3.3 ml/cm2 vs. 2.8±4.0 ml/cm2 , p=0.006). No significant differences were observed in the rate of complications between the groups. Conclusions: The findings suggest that the monopolar electrocoagulation created with an internally-cooled RF electrode considerably reduces intraoperative blood loss during liver resection., This work was supported by a medical research grant from the Spanish Government (FIS PI080934) and by Spanish "Programa Estatal de Investigacion, Desarrollo e Innovacion Orientada a los Retos de la Sociedad" under Grant TEC2014-52383-C3-R (TEC2014-52383-C3-1-R and TEC2014-52383-C3-3-R).

The purpose of this study was to evaluate plasma concentrations and pharmacokinetic parameters of buprenorphine in dogs following intravenous (IV) administration of clinical doses of the opioid. An IV bolus of 0.02 mg/kg buprenorphine was administered to six healthy Beagles and blood samples were collected through a jugular catheter before and at 1, 5, 10, 15, 20, 30 and 45 min, and 1, 2, 4, 6, 8 and 12 h after administration. Plasma buprenorphine concentrations, measured using a commercial radioimmunoassay (RIA), decreased following a three-exponential curve. The two distribution and the elimination half-lives were 2.9 ± 1.8 min, 16.5 ± 3.7 min, and 266.6 ± 82.0 min, respectively; the clearance was 329.6 ± 62.2 mL/min, and the steady state volume of distribution was 83.7 ± 26.5 L. The results demonstrated the feasibility of the RIA assay to analyse buprenorphine in dog plasma samples. Following IV administration buprenorphine showed a three-compartment kinetic profile, as has been described previously in humans, rabbits and cats. The relationship between plasma concentrations and dynamic effects in dogs remains to be established.

Etomidate is an intravenous (IV) hypnotic agent characterised by its cardiovascular stability. Although etomidate has been satisfactorily used in veterinary and human obstetrics, little is known about its effects on the fetus. This study determined the cardiovascular and acid-base effects of etomidate administration in the pregnant ewe and her fetus. The effects of etomidate were evaluated in two separate studies. In the first study, etomidate was administered as a 1mg/kg IV bolus; in the second, the drug was administered as a continuous infusion of 100 microg/kg/min for 1h, preceded by a 1mg/kg IV bolus. Etomidate administration did not depress cardiovascular function in the pregnant ewe or fetus. When administered as a continuous infusion, maternal heart rate and blood pressure increased during the second half of the infusion and the initial stages of recovery. Acid-base alterations led to transient but slight respiratory depression in both mother and fetus, probably reflecting the combined effects of etomidate on respiration and the positioning of the animal.

Etomidate (ETO) is a short-acting intravenous (IV) anaesthetic characterised by cardiopulmonary stability and favourable pharmacokinetics. Although ETO has been used satisfactorily in obstetrical anaesthesia, little is known about placental transfer and the drug’s pharmacokinetics in the fetus. Placental transfer in pregnant ewes has been evaluated following the administration of an IV bolus of 1 mg/kg ETO; and after a 1-h infusion of 100 μg/kg min −1 ETO preceded by an IV bolus of 1 mg/kg. In ewes, ETO concentration and AUC were higher than those found in fetuses. After the ETO bolus dose, the fetus:ewe AUC ratio was 0.45 ± 0.32, and the mean residence time (MRT) was 20 ± 7 min for dams and 22 ± 3 min for the fetuses. After ETO infusion, the AUC ratio was 0.37 ± 0.08, and MRT was 46 ± 12 min for ewes and 46 ± 22 min for fetuses. Although ETO crosses the placenta very rapidly and reaches the fetus in high amounts, a certain placental barrier effect limits its transfer. There is no evidence of cumulative effects of the drug in the fetus as fetal ETO elimination was as rapid as in the dam.

Purpose: To assess the capacity of two methods of surgical pancreatic stump closure in terms of reducing the risk of pancreatic fistula formation (POPF): radiofrequencyinduced heating versus mechanical stapler. Materials and Methods: Sixteen pigs underwent a laparoscopic transection of the neck of the pancreas. Pancreatic anastomosis was always avoided in order to work with an experimental model prone to pancreatic fistula formation (POPF). Pancreatic stump closure was conducted either by stapler (ST Group, n=8) or radiofrequency energy (RF Group, n=8). Both groups were compared for incidence of POPF and histopathologic alterations of the pancreatic remnant. Results: Six animals (75%) in the ST Group and one (14%) in the RF Group were diagnosed of POPF (p=0.019). One animal in the RF Group and three animals in the ST Group had a pseudocyst in close contact with both pancreas stumps. On day 30 postoperative, almost complete atrophy of the exocrine distal pancreas was observed when the main pancreatic duct was efficiently sealed. Conclusions: Our findings suggest that RF-induced heating is more effective at closing the pancreatic stump than mechanical stapler and leads to the complete atrophy of the distal remnant pancreas., This work was supported by the Spanish 'Programa Estatal de Investigacion, Desarrollo e Innovacion Orientada a los Retos de la Sociedad' under grant TEC2014-52383-C3-R (TEC2014-52383-C3-3-R). F.B., R.Q. and E.B. declare stock ownership in Apeiron Medical S.L., a company that has a license for the patent US 8.303.584.B2, on which the device tested in this study is based. The other authors report no conflict of interests or financial ties to disclose. The authors alone are responsible for the content and writing of the paper.

Background: Pancreatic duct ligation (PDL) has been used as a model of chronic pancreatitis and as a model to increase β-cell mass. However, studies in mice have demonstrated acinar regeneration after PDL, questioning the long-term validity of the model. We aim to elucidate whether RF-assisted transection (RFAT) of the main pancreatic duct is a reliable PDL model, both in short (ST, 1-month) and long-term (LT, 6-months) follow-ups. Methods: Eleven pigs were subjected to RFAT. Biochemical (serum/peripancreatic amylase and glucose) and histological changes (including a semiautomatic morphometric study of over 1000 images/pancreas and IHC analysis) were evaluated after ST or LT follow-up and also in fresh pancreas specimens that were used as controls for 1 (n=4) and 6 months (n=6). Results: The distal pancreas in the ST was characterized by areas of acinar-to-ductal metaplasia (56%) which were significantly reduced at LT (21%) by fibrotic replacement and adipose tissue. The endocrine mass showed a normal increase. Conclusion: RFAT in the pig seems to be an appropriate PDL model without restoration of pancreatic drainage or reduction of endocrine mass., "This work was supported by the Spanish "Plan Estatal de Investigacion, Desarrollo e Innovacion Orientada a los Retos de la Sociedad" under Grant TEC2014-52383-C3 (TEC2014-52383-C3-3-R). RQ, EB, FB declare stock ownership in Apeiron Medical S.L., a company that has a license for the patent US 8.303.584.B2, on which the device have been employed.

Propofol is an injectable anaesthetic that is currently used both in veterinary and human medicine for the induction and maintenance of anaesthesia. Although little is known about the pharmacokinetics of propofol in fetuses, it is widely used in obstetric procedures, particularly in caesarean section. This study determines the pharmacokinetics of propofol in pregnant ewes in the last third of pregnancy, and placental transfer and pharmacokinetics in fetuses after the administration of a 6 mg/kg intravenous (i.v.) bolus (phase 1) or a 6 mg/kg i.v. bolus followed by continued infusion of 0.4 mg/kg/min. In ewes, the area under the blood concentration–time curve (AUC) and C max (8.6 mg h/mL and 9.5 mg/mL, respectively) was higher than those of the fetus (1.6 mg h/mL and 1.19 mg/mL, respectively). The mean half-life was 0.5 h in the dam and 1.1 h in the fetus.

Electroporation based treatments typically consist in applying high voltage dc pulses. As an undesired side effect, these dc pulses cause electrical stimulation of excitable tissues such as motor nerves. In the present in vivo study, it was explored the use of bursts of sinusoidal voltage in the frequency range from 50 kHz to 2 MHz to induce irreversible electroporation (IRE) whilst avoiding neuromuscular stimulation. Series of 100 dc pulses or sinusoidal bursts, both with an individual duration of 100 μs, were delivered to rabbit liver through thin needles in a monopolar electrode configuration and thoracic movements were recorded with an accelerometer. Tissue samples were harvested three hours after treatment and later postprocessed to determine the dimensions of the IRE lesions. Thermal damage due to Joule heating was ruled out via computer simulations. Sinusoidal bursts with a frequency equal or above 100 kHz did not cause thoracic movements and induced lesions equivalent to those obtained with conventional dc pulses when the applied voltage amplitude was sufficiently high. IRE efficacy dropped with increasing frequency. For 100 kHz bursts, it was estimated that the electric field threshold for IRE is about 1.4 kV/cm whereas that of dc pulses is about 0.5 kV/cm. This work was supported by the Ministry of Economy and Competitiveness of Spain through the grant TEC2014-52383-C3-R (TEC2014-52383-C3-2-R and TEC2014-52383-C3-3-R).

The aim of our study was to assess whether the heat sink effect inside the portal vein could protect its wall from thermal damage close to an internally cooled electrode during RF-assisted resection. Computer simulations were carried out to study the effect of different factors such as device-tissue contact, vessel position and vessel-device distance on temperature distributions and thermal lesion shapes near the portal vein. Our results suggest that the heat sink effect could protect the portal vein wall for distances equal to or greater than 5 mm, regardless of its position and distance with respect to the RF-based device.

Summary Metastases in the liver frequently grow as scattered tumor nodules which neither can be removed by surgical resection nor focally ablated. Previously, we have proposed a novel technique based on irreversible electroporation which may be able to simultaneously treat all nodules in the liver while sparing healthy tissue. The proposed technique requires increasing the electrical conductivity of healthy liver by injecting a hypersaline solution through the portal vein. Aiming to assess the capability of increasing the global conductivity of the liver by means of hypersaline fluids, here it is presented a mathematical model which estimates the NaCl distribution within the liver and the resulting conductivity change. The model fuses well-established compartmental pharmacokinetic models of the organ with saline injection models employed for resuscitation treatments and it considers changes in sinusoidal blood viscosity due to the hypertonicity of the solution. Here it is also described a pilot experimental study in pigs in which different volumes of NaCl 20% (from 100 to 200 ml) were injected through the portal vein at different flow rates (from 53 to 171 ml/min). The in vivo conductivity results fit those obtained by the model, both quantitatively and qualitatively, being able to predict the maximum conductivity with a 14.6% average relative error. The maximum conductivity value was 0.44 S/m which corresponds to increasing four times the mean basal conductivity (0.11 S/m). The results suggest that the presented model is well suited for predicting on liver conductivity changes during hypertonic saline injection. This article is protected by copyright. All rights reserved.

The purpose of this randomised trial was to compare the effects of alfaxalone and propofol on tear production (STT-1), intraocular pressure (IOP) and globe position (GP) in healthy dogs. Fourteen Beagles were randomly divided into two groups; dogs in one group received alfaxalone (3 mg/kg) (n=7) and dogs in the other group received propofol (6 mg/kg) (n=7), both administered intravenously. IOP and GP were evaluated at basal time (Tb) and T2,5,10,15,20,2530 (minutes after complete drug administration). STT-1 was evaluated at Tb and T10,2030. STT-1 and IOP results were analysed using analysis of variance and GP was analysed using the likelihood ratio χ(2) test. Dogs in the alfaxalone group showed a significant reduction in STT-1 at T1020 (P0.05), while the propofol group did not show statistically significant differences in this parameter over time. Both anaesthetic drugs produced a transient elevation of IOP at T2 (P0.05), which then decreased (P0.01). While alfaxalone caused a ventral globe deviation that lasted from T2 to T10 (P0.05) and was fully recovered at T30, propofol induced a ventral globe deviation from T2 to T5 (P0.05), being restored at T20. These results suggest that both alfaxalone and propofol can be safely used for intraocular surgery, as they significantly reduce IOP. Furthermore, anaesthetic induction with propofol would be especially recommended for dogs with tear deficiencies.

To assess by means of computer simulations whether the heat sink effect inside a large vessel (portal vein) could protect the vessel wall from thermal damage close to an internally cooled electrode during radiofrequency (RF)-assisted resection.First,in vivo experiments were conducted to validate the computational model by comparing the experimental and computational thermal lesion shapes created around the vessels. Computer simulations were then carried out to study the effect of different factors such as device-tissue contact, vessel position, and vessel-device distance on temperature distributions and thermal lesion shapes near a large vessel, specifically the portal vein.The geometries of thermal lesions around the vessels in the in vivo experiments were in agreement with the computer results. The thermal lesion shape created around the portal vein was significantly modified by the heat sink effect in all the cases considered. Thermal damage to the portal vein wall was inversely related to the vessel-device distance. It was also more pronounced when the device-tissue contact surface was reduced or when the vessel was parallel to the device or perpendicular to its distal end (blade zone), the vessel wall being damaged at distances less than 4.25 mm.The computational findings suggest that the heat sink effect could protect the portal vein wall for distances equal to or greater than 5 mm, regardless of its position and distance with respect to the RF-based device.

Amyotrophic lateral sclerosis (ALS) is an adult onset neurodegenerative disease that causes progressive paralysis and death due to degeneration of motoneurons in spinal cord, brainstem and motor cortex. Nowadays, there is no effective therapy and patients die 2-5 years after diagnosis. Resveratrol (trans-3,4',5-trihydroxystilbene) is a natural polyphenol found in grapes, with promising neuroprotective effects since it induces expression and activation of several neuroprotective pathways involving Sirtuin1 and AMPK. The objective of this work was to assess the effect of resveratrol administration on SOD1(G93A) ALS mice. We determined the onset of symptoms by rotarod test and evaluated upper and lower motoneuron function using electrophysiological tests. We assessed the survival of the animals and determined the number of spinal motoneurons. Finally, we further investigated resveratrol mechanism of action by means of western blot and immunohistochemical analysis. Resveratrol treatment from 8 weeks of age significantly delayed disease onset and preserved lower and upper motoneuron function in female and male animals. Moreover, resveratrol significantly extended SOD1(G93A) mice lifespan and promoted survival of spinal motoneurons. Delayed resveratrol administration from 12 weeks of age also improved spinal motoneuron function preservation and survival. Further experiments revealed that resveratrol protective effects were associated with increased expression and activation of Sirtuin 1 and AMPK in the ventral spinal cord. Both mediators promoted normalization of the autophagic flux and, more importantly, increased mitochondrial biogenesis in the SOD1(G93A) spinal cord. Taken together, our findings suggest that resveratrol may represent a promising therapy for ALS.

The purpose of the study was to evaluate the effect of preoperative administration of meloxicam, a non-steroidal anti-inflammatory drug used for pain control, on primary haemostasis in dogs. Twenty healthy female dogs undergoing elective ovariohysterectomy were enrolled in the study. Sixty minutes before pre-anaesthesia, a single dose of meloxicam (0.2 mg/kg) was randomly administered intravenously (IV) to 10 dogs (treatment group) while control dogs received an equivalent volume of saline solution IV. Platelet aggregation, buccal mucosa bleeding time, platelet count and haematological indices were measured at 0, 1, 6 and 24 h after administration of meloxicam. Since significant differences between groups were not observed for any of the measured parameters, preoperative administration of meloxicam may be used for pain control before elective ovariohysterectomy in healthy dogs, without compromising primary haemostasis.

To investigate the diagnostic and prognostic value over time of plasma iron compared with the inflammatory markers albumin, C-reactive protein (CRP), and fibrinogen in dogs with systemic inflammatory response syndrome (SIRS).Prospective observational study of sequentially enrolled dogs.ICU of a veterinary teaching hospital.One hundred and sixteen client-owned dogs: 54 dogs with SIRS or sepsis, 42 with focal inflammation, and 20 clinically healthy dogs.Blood samples were obtained on admission in all study groups, and then on alternate days until discharge or death in both inflammation groups. On admission, dogs with SIRS had significantly lower plasma iron (65 ± 5.8 μg/dL, P = 0.001) concentrations than dogs with focal inflammation (89.5 ± 6.2 μg/dL, P = 0.001). Plasma iron, albumin, and CRP effectively discriminated the SIRS/sepsis group from those presenting with focal inflammation with areas under the curve for the receiver operating curves of 0.679, 0.834, and 0.704, respectively. The admission values for these variables did not discriminate survivors from nonsurvivors within the SIRS/sepsis group. However, the magnitude of increase in iron concentration and the decrease in CRP concentration from admission to hospital discharge was higher in survivors than in nonsurvivors within the SIRS/septic group (22.8 vs. 2.51 μg/dL, respectively, P = 0.021 for iron; -67.1 vs. -4.1 mg/L, respectively, P = 0.002 for CRP), resulting in iron and CRP concentrations at hospital discharge for survivors similar to those in the focal inflammation group.Hypoferremia is a sensitive marker of systemic inflammation in dogs. In this study, the increase in iron concentrations during the hospitalization period of SIRS/septic dogs was associated with a better prognosis, suggesting that plasma iron in combination with CRP and albumin concentrations might be used to monitor dogs with inflammatory disease processes.

Introduction Umbilical artery (UA) hemodynamics reflect blood flow and vascular resistance in the placental circulation. We examined non-invasively the hemodynamic effects of propofol, etomidate, and alphaxalone on the placental circulation of a sheep model by means of UA Doppler ultrasonography. Methods Eleven sheep fetuses were examined at 90–109 days of gestation. UA Doppler ultrasound was performed before and after administration of a single intravenous bolus of propofol, etomidate, or alphaxalone. UA Doppler velocities (peak systolic velocity, end diastolic velocity, and mean velocity), vascular indices (pulsatility index, resistance index, and S/D ratio), blood flow, and fetal heart rate were recorded during the experimental period and UA Doppler waveforms were characterized. Results A laminar, parabolic, low resistance flow was observed in the UA of the sheep fetuses. No statistically significant changes were observed in the UA Doppler waveforms or in the UA Doppler hemodynamics after anesthesia induction. Discussion Changes in placental vascular resistance may alter the corresponding UA Doppler waveforms. When resistance in the fetal placenta increases, blood flow in the UA becomes more pulsatile. In the present study, umbilical arteries showed a parabolic flow with low resistance in all cases, as it occurs in normal human pregnancy. The administration of these anesthetics did not cause abnormalities in the normal UA Doppler pattern, inducing no changes in the resistance of the placenta in any case. Conclusion These results suggest that intravenous anesthetic induction with propofol, etomidate, or alphaxalone does not cause significant detrimental effects on the placental circulation of the pregnant ewe.

The objective of this study was to determine the pharmacodynamics effects of the anaesthetic alfaxalone in 2-hydroxypropyl-β-cyclodextrin in pregnant sheep after the intravenous injection of a 2 mg/kg weight dose. Six pregnant Ripollesa sheep, weighing 47.1 ± 4.4 kg, were used. Twenty-four hours after instrumentation, sheep were anaesthetized with intravenous alfaxalone in cyclodextrin. Time to standing from anaesthesia was 30.0 ± 10.81 min. Foetal heart rate increased significantly during the first 5 min after alfaxalone administration. Significant differences were observed in maternal diastolic arterial blood pressure between minute 10 and minutes 90, 120, 150, 180, 210, and 240. Significant differences were observed for foetal systolic arterial blood pressure between 5 and 30 min after alfaxalone administration. Significant differences in foetal pH were detected during the entire study period, whereas maternal pH returned to baseline values by 60 min after alfaxalone administration. The present study indicated that alfaxalone in 2-hydroxypropyl-β-cyclodextrin administered as an intravenous bolus at 2 mg/kg body weight produced minimal adverse effects and an uneventful recovery from anaesthesia in pregnant sheep and their foetus.

[EN] Postoperative pancreatic fistula (PPF) is the most frequent and serious complication after laparoscopic distal pancreatectomy (LDP). Our goal was to compare the performance, in terms of PPF prevention, and safety of a radiofrequency (RF)-assisted transection device versus a stapler device in a porcine LDP model. Thirty-two animals were randomly divided into two groups to perform LDP using a RF-assisted device (RF group; n = 16) and stapler device (ST group; n = 16) and necropsied 4 weeks after surgery. The primary endpoint was the incidence of PPF. Secondary endpoints were surgery/transection time, intra/postoperative complications/deaths, postoperative plasmatic amylase and glucose concentration, peritoneal liquid amylase and interleukin 6 (IL-6) concentrations, weight variations, and histopathological changes. Two clinical and one biochemical PPF were observed in the ST and RF groups respectively. Peritoneal amylase concentration was significantly higher in the RF group 4 days after surgery, but this difference was no longer present at necropsy. Both groups presented a significant decrease in peritoneal IL-6 concentration during the postoperative follow-up, with no differences between the groups. RF group animals showed a higher postoperative weight gain. In the histopathological exam, all RF group animals showed a common pattern of central coagulative necrosis of the parenchymal surface, surrounded by a thick fibrosis, which sealed main and secondary pancreatic ducts and was not found in ST group. The fibrosis caused by an RF-assisted device can be at least as safe and effective as stapler compression to achieve pancreatic parenchyma sealing in a porcine LDP model., This work received financial support from the Spanish "Plan Nacional de I+D+I del Ministerio de Ciencia e Innovacion," Grant No. TEC2011-27133-C02-02.

Purpose: The aim of this study was to assess the feasibility of a hybrid ablative technique based on applying electroporation (EP) pulses just before conducting radiofrequency ablation (RFA). The rationale was that the EP-induced reduction in blood perfusion could be sufficient to reduce the thermal sink effect and hence to increase the coagulation volume in comparison to that created exclusively by RFA. Materials and methods: A modelling study and in vivo experimental study were used. A Cool-tip RF applicator was used both for EP and RFA. Results: Overall, the results did not show any synergy effect from using the hybrid technique. Applying EP pulses prior to RFA did not increase the coagulation zone obtained and the lesions were almost identical. Additional computer simulations provided an explanation for this; the effect of reducing blood perfusion by thermal damage during RFA completely masks the effect of reducing blood perfusion by EP. This is because both thermal damage and EP affect the same zone, i.e. the tissue around the electrode. Conclusions: Our computer modelling and in vivo experimental findings suggest that the combination of EP and RFA with monopolar applicators does not provide an additional benefit over the use of RFA alone., This work received financial support from the Spanish 'Plan Nacional de I + D + I del Ministerio de Ciencia e Innovacion' Grant nos. TEC2011-27133-C02-01,02 and TEC2010-17285, from the European Commission through the Marie Curie IRG grant TAMIVIVE (256376) and from the Universitat Politecnica de Valencia (INNOVA11-01-5502; and PAID-06-11 Ref. 1988). The linguistic revision of this paper was funded by the Universitat Politecnica de Valencia, Spain. The authors alone are responsible for the content and writing of the paper.

Intra-abdominal pressure (IAP) and abdominal perfusion pressure (APP) have shown clinical relevance in monitoring critically ill human beings submitted to abdominal surgery. Only a few studies have been performed in veterinary medicine. The aim of this study was to assess how pregnancy and abdominal surgery may affect IAP and APP in healthy cats. For this purpose, pregnant (n=10) and non-pregnant (n=11) queens undergoing elective spaying, and tomcats (n=20, used as controls) presented for neutering by scrotal orchidectomy were included in the study. IAP, mean arterial blood pressure (MAP), APP, heart rate and rectal temperature (RT) were determined before, immediately after, and four hours after surgery. IAP increased significantly immediately after abdominal surgery in both female groups when compared with baseline (P