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Abstract Purpose Volumetric assessment of the liver regularly yields discrepant results between pre- and intraoperatively determined volumes. Nevertheless, the main factor responsible for this discrepancy remains still unclear. The aim of this study was to systematically determine the difference between in vivo CT-volumetry and ex vivo volumetry in a pig animal model. Material and Methods Eleven pigs were studied. Liver density assessment, CT-volumetry and water displacement volumetry was performed after surgical removal of the complete liver. Known possible errors of volume determination like resection or segmentation borders were eliminated in this model. Regression analysis was performed and differences between CT-volumetry and water displacement determined. Results Median liver density was 1.07 g/ml. Regression analysis showed a high correlation of r2 = 0.985 between CT-volumetry and water displacement. CTvolumetry was found to be 13% higher than water displacement volumetry (p < 0.0001). Conclusion In this study the only relevant factor leading to the difference between in vivo CT-volumetry and ex vivo water displacement volumetry seems to be blood perfusion of the liver. The systematic difference of 13 percent has to be taken in account when dealing with those measures.

Liver cells cultured in 3D bioreactors is an interesting option for temporary extracorporeal liver support in the treatment of acute liver failure and for animal models for preclinical drug screening. Bioreactor capacity to eliminate drugs is generally used for assessing cell metabolic competence in different bioreactors or to scale-up bioreactor design and performance for clinical or preclinical applications. However, drug adsorption and physical transport often disguise the intrinsic drug biotransformation kinetics and cell metabolic state. In this study, we characterized the intrinsic kinetics of lidocaine elimination and adsorption by porcine liver cells cultured in 3D four-compartment hollow fiber membrane network perfusion bioreactors. Models of lidocaine transport and biotransformation were used to extract intrinsic kinetic information from response to lidocaine bolus of bioreactor versus adhesion cultures. Different from 2D adhesion cultures, cells in the bioreactors are organized in liver-like aggregates. Adsorption on bioreactor constituents significantly affected lidocaine elimination and was effectively accounted for in kinetic analysis. Lidocaine elimination and cellular monoethylglicinexylidide biotransformation featured first-order kinetics with near-to-in vivo cell-specific capacity that was retained for times suitable for clinical assist and drug screening. Different from 2D cultures, cells in the 3D bioreactors challenged with lidocaine were exposed to close-to-physiological lidocaine and monoethylglicinexylidide concentration profiles. Kinetic analysis suggests bioreactor technology feasibility for preclinical drug screening and patient assist and that drug adsorption should be accounted for to assess cell state in different cultures and when laboratory bioreactor design and performance is scaled-up to clinical use or toxicological drug screening.

Systemic diseases characterized by elevated levels of C-reactive protein (CRP), such as sepsis or systemic inflammatory response syndrome, are usually associated with hardly controllable haemodynamic instability. We therefore investigated whether CRP itself influences blood pressure and heart rate. Immediately after intravenous injection of purified human CRP (3.5 mg CRP/kg body weight) into anesthetized rabbits, blood pressure dropped critically in all animals, while control animals injected with bovine serum albumin showed no response. Heart rate did not change in either group. Approaching this impact on a cellular level, we investigated the effect of CRP in cell lines expressing adrenoceptors (CHO-α1A and DU-145). CRP caused a Ca 2+ signaling being dependent on the CRP dose. After complete activation of the adrenoceptors by agonists, CRP caused additional intracellular Ca 2+ mobilization. We assume that CRP interacts with hitherto unknown structures on the surface of vital cells and thus interferes with the desensitization of adrenoceptors., (Copyright © 2020 Bock, Vogt, Mattecka, Yapici, Brunner, Fimpel, Unger and Sheriff.)

Hepatocyte transplantation (HcTx) is a promising approach for the treatment of metabolic diseases in newborns and children. The most common application route is the portal vein, which is difficult to access in the newborn. Transfemoral access to the splenic artery for HcTx has been evaluated in adults, with trials suggesting hepatocyte translocation from the spleen to the liver with a reduced risk for thromboembolic complications. Using juvenile Göttingen minipigs, we aimed to evaluate feasibility of hepatocyte transplantation by transfemoral splenic artery catheterization, while providing insight on engraftment, translocation, viability, and thromboembolic complications. Four Göttingen Minipigs weighing 5.6 kg to 12.6 kg were infused with human hepatocytes (two infusions per cycle, 1.00E08 cells per kg body weight). Immunosuppression consisted of tacrolimus and prednisolone. The animals were sacrificed directly after cell infusion ( n =2), 2 days ( n =1), or 14 days after infusion ( n =1). The splenic and portal venous blood flow was controlled via color-coded Doppler sonography. Computed tomography was performed on days 6 and 18 after the first infusion. Tissue samples were stained in search of human hepatocytes. Catheter placement was feasible in all cases without procedure-associated complications. Repetitive cell transplantations were possible without serious adverse effects associated with hepatocyte transplantation. Immunohistochemical staining has proven cell relocation to the portal venous system and liver parenchyma. However, cells were neither present in the liver nor the spleen 18 days after HcTx. Immunological analyses showed a response of the adaptive immune system to the human cells. We show that interventional cell application via the femoral artery is feasible in a juvenile large animal model of HcTx. Moreover, cells are able to pass through the spleen to relocate in the liver after splenic artery infusion. Further studies are necessary to compare this approach with umbilical or transhepatic hepatocyte administration.

Acute kidney injury (AKI) corrupts the outcome of about 50% of all critically ill patients. We investigated the possible contribution of the pathology acidemia on the development of AKI. Pigs were exposed to acidemia, acidemia plus hypoxemia or a normal acid-base balance in an experimental setup, which included mechanical ventilation and renal replacement therapy to facilitate biotrauma caused by extracorporeal therapies. Interestingly, extensive histomorphological changes like a tubular loss of cell barriers occurred in the kidneys after just 5 hours exposure to acidemia. The additional exposure to hypoxemia aggravated these findings. These 'early' microscopic pathologies opposed intra vitam data of kidney function. They did not mirror cellular or systemic patterns of proinflammatory molecules (like TNF-α or IL 18) nor were they detectable by new, sensitive markers of AKI like Neutrophil gelatinase-associated lipocalin. Instead, the data suggest that the increased renal proton excretion during acidemia could be an 'early' first hit in the multifactorial pathogenesis of AKI., Competing Interests: All authors declare no further competing interests, especially concerning employment, consultancy, patents, products in development or marketed products. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

Over the past 50 years, image-guided procedures have been established for a wide range of applications. The development and clinical translation of new treatment regimens necessitate the availability of suitable animal models. The juvenile Göttingen minipig presents a favourable profile as a model for human infants. However, no information can be found regarding the vascular system of juvenile minipigs in the literature. Such information is imperative for planning the accessibility of target structures by catheterization. We present here a complete mapping of the arterial system of the juvenile minipig based on contrast-enhanced computed tomography. Four female animals weighing 6.13 ± 0.72 kg were used for the analyses. Imaging was performed under anaesthesia, and the measurement of the vascular structures was performed independently by four investigators. Our dataset forms a basis for future interventional studies in juvenile minipigs, and enables planning and refinement of future experiments according to the 3R (replacement, reduction and refinement) principles of animal research.

Investigation: A novel active wound dressing (AWD) concept based on a microporous hollow fiber membrane network was investigated in an animal model. It provides a local solution-perfused environment for regenerative cell nutrition, wound irrigation, debris removal, electrolyte balancing, pH regulation, and topical antibiosis. The device is capable of supplying soluble factors, as tested experimentally for the recombinant human growth and differentiation factor-5 (rhGDF-5)., Methods: Following in vitro studies for rhGDF-5 using primary human keratinocytes and dermal fibroblasts, we employed a porcine partial thickness wound model with five distinct wounds on each back of n=8 pigs. Four wound groups were perfused differently over 9 days and compared with a negative control wound without perfusion: (1) 1% trehalose solution, pH 5.5; (2) rhGDF-5 (150 ng/ml) in 1% trehalose solution, pH 5.5; (3) nutrition solution; and (4) rhGDF-5 (150 ng/ml) in nutrition solution with 1% trehalose, pH 5.5., Results: Promoted wound healing was observed within group 1 and more pronounced within group 2. Groups 3 and 4, with nutrition solution, showed significant adverse effects on wound healing (p<0.05)., Conclusions: The investigated AWD concept appears to be an interesting therapeutic tool to study further wound healing support. Additionally, topical application of rhGDF-5 could be promising., (Copyright © 2014 Elsevier Ltd and ISBI. All rights reserved.)

Background: There is substantial evidence that C-reactive protein (CRP) mediates secondary damage of the myocardium after acute myocardial infarction (AMI). The aim of this animal trial in pigs was to specifically deplete CRP from porcine plasma after AMI and to study possible beneficial effects of the reduced CRP concentration on the infarcted area., Methods: Ten pigs received balloon catheter-induced myocardial infarction. CRP was depleted from five animals utilizing a new specific CRP-adsorber, five animals served as controls. The area of infarction was analyzed by cardiovascular magnetic resonance imaging on day 1 and day 14 after AMI. Porcine CRP levels were determined by ELISA., Results: CRP-apheresis resulted in a mean reduction of the CRP levels up to 48.3%. The area of infarction was significantly reduced by 30 ± 6% (P = 0.003) within 14 days in the treatment group, whereas it increased by 19 ± 11% (P = 0.260) in the controls. Fourteen days after infarction, the infarcted area revealed compact, transmural scars in the controls, whereas animals receiving CRP-apheresis showed spotted scar morphology. In the interventional group, a significantly higher left ventricular ejection fraction (LVEF) was observed after 14 days as compared to the controls (57.6 ± 2.4% vs. 46.4 ± 2.7%; P = 0.007)., Conclusions: In a pig model for AMI, we observed that selective CRP-apheresis significantly reduces CRP levels and the volume of the infarction zone after AMI. Additionally, it changes the morphology of the scars and preserves cardiac output (LVEF)., (© 2014 Wiley Periodicals, Inc.)

Experimental data indicate that hypercapnic adidosis has anti-inflammatory effects. These anti-inflammatory effects may even be a beneficial property in case of low tidal volume ventilation with consecutive hypercapnic acidosis. It is unclear whether these anti-inflammatory effects predominate in critically ill patients who suffer from multiple pro- and anti-inflammatory insults like extracorporeal organ support (pro-inflammatory), metabolic acidosis (pro- and anti-inflammatory), as well as hypoxia (pro-inflammatory). Eighteen pigs were randomized into three groups, mechanically ventilated and connected to a continuous veno-venous hemofiltration (CVVH) as pro-inflammatory insult. A reference group with normal acid-base state obtained normoventilation; a normoxemic acidemia group obtained normoxemic, mixed acidemia due to infusion of lactic and hyperchloremic acid and low tidal volume ventilation, and in a hypoxemic acidemia group the mixed acidemia was paralleled by hypoxemia. Lung histology including pulmonary leukocyte invasion, blood gases, blood cell counts, and hemodynamics were examined. The histological examination of the lungs of acidemic pigs showed a suppressed invasion of leukocytes and thinner alveolar walls compared with normoventilated and with hypoxemic pigs. Enhanced congestion and alveolar red blood cells (RBCs) combined with an increase of the pulmonary artery pressure were observed in acidemic pigs in comparison with the reference group. Normoxemic acidemia reduced the pro-inflammatory reaction to the CVVH and mechanical ventilation in the ventilated lung areas in the form of pulmonary leukocyte invasion. However, this did not result in reduced scores for lung injury. Instead, an increased score for criteria which represent lung injury (congestion and alveolar RBCs) was observed in acidemic pigs., (© 2013 Wiley Periodicals, Inc. and International Center for Artificial Organs and Transplantation.)

Intensive therapy demanding diseases (organ failure or sepsis) are assumed to be the etiology behind a decreased biocompatibility of extracorporeal systems for renal replacement therapy (RRT). There are also potential interactions between different components of the overall therapy. Volume substitutes are known to influence hemorheology and coagulation. To define a potential net effect of volume substitutes on the hemocompatibility of an RRT, we chose an animal model without interfering pathophysiologies. According to the problem of early filter failure and coagulation disorders in critically ill patients, we focused on the hypothesized interaction between RRT and different volume substitutes with respect to blood cell counts, coagulation parameters and required heparin dose. Forty-eight pigs were assigned to four groups of fluid therapy with either normal saline (NaCl), 6%HES130kD/0.4 (HES130), 6%HES200kD/0.5 (HES200) or 4%gelatin (GEL). Six pigs of each fluid group underwent continuous venovenous hemofiltration (CVVH), the remaining six served as the control group. Anticoagulation was performed with continuous heparin infusion. CVVH was run in a recirculation-mode for 4.5 h to force hemocompatibility reactions, thereafter in a standard-mode for 2 h. During the CVVH-treatment GEL reduced platelet counts and fibrinogen concentration and additionally lowered ATIII levels. Heparin requirements did not differ between different volume substitutes or CVVH and control groups. Severe pathophysiologies are not the only reason for a reduced hemocompatibility of CVVH treatment. Interaction of a particular volume substitute with CVVH should be considered when interpreting study results and evolving new strategies., (© 2013 The Authors. Therapeutic Apheresis and Dialysis © 2013 International Society for Apheresis.)

Background: A standardized large animal model for controlled ICP manipulation within a relevant range and repetitive ICP measurements is missing. We sought to develop such a model on the base of controlled IPP changes induced by capnoperitoneum., Methods: We utilized six female pigs (mean body weight 59.5 ± 18.4 kg) for experiments. A ventricular catheter connected with a burr hole reservoir was implanted. ICP was measured directly as cm H2O within a riser tube after percutaneous cannulation of the reservoir. A noninvasive intraperitoneal pressure (IPP) measurement was established (intravesical). Animals were placed in lateral position and a capnoperitoneum was induced. Measurements of ICP, IPP, MAP and respiratory parameters were performed at baseline IPP and after CO2 insufflation to IPP levels of 20 and 30 mmHg., Results: Baseline IPP in lateral position referenced to median line was 9.8 (±2) mm Hg, while corresponding ICP was 10 (±2.2) mm Hg. After IPP elevation to 20 mmHg, ICP increased to 18.8 (±1.9) mm Hg. At 30 mmHg IPP, ICP increased to 22.8 (±2.8) mm Hg. Except peak airway pressure, all other parameters were kept constantly. Mean ICP variation in the individual subject was 13.4 (±2.5) mm Hg, while a ICP range from minimum 9 to maximum 31 mmHg was documented., Conclusions: We report a large animal model that allows (1) repeated measurement of the ICP and (2) manipulation of the ICP within a large pressure range by controlled IPP changes due to capnoperitoneum.

In intensive care medicine, convection-based apheresis is of growing interest. Applying extracorporeal systems in the critically ill patient can cause severe complications like nosocomial infections and bleeding, which can be worsened or even initialized by the anticoagulation protocol used. Furthermore, the filter modules (hemo- and plasmafilters) often tend to a fast blockage. A decrease in sieving performance due to membrane fouling may be tolerable for some time, but the complete blockage of high percentages of hollow fibers, which is named "clotting," often requires the immediate exchange of the filter. Extracorporeal detoxification and high clearance renal replacement regimes both require high blood flow and filtration rates. As a consequence, filter clotting and anticoagulation-associated bleeding are the most sensitive aspects in these applications. We were interested in the paradox phenomenon of the parallel occurrence of intra vitam bleeding and filter clotting in critically ill patients. Through stepwise investigations based on in vitro and animal experiments, we identified a stasis of blood flow followed by blood cell sedimentation and aggregation ("clogging") as the main factor of hollow fiber blockage in hemo- and plasma filters. As a result, various aspects which increase the risk of stasis inside the hollow fibers were investigated, for example, patient's hemorheology, configuration of an extracorporeal treatment system including interaction of catheter features with the filtration procedure, and basic therapeutic approaches such as colloidal volume substitutes and tolerated acidosis. Finally, an etiological triad for the blockage of hollow fibers due to filter clogging and consecutive filter failure was formed., (© 2013, Copyright the Authors. Artificial Organs © 2013, International Center for Artificial Organs and Transplantation and Wiley Periodicals, Inc.)

Exhausting physical exercise and insufficient nutritional intake impairing immunological and neuro-endocrine pathways are the most discussed issues in research on overtraining syndrome (OTS). Interestingly, depletion of the total body sodium (Na(+))-content which occurs in case of various diseases with completely different aetiologies is associated with a symptom pattern strikingly comparable to overreaching (OR) and/or OTS. The transient dilution based hyponatraemia gained attention due to its impact on reduced performance and the death of various endurance athletes. But the stepwise depletion of the total body (tissue) Na(+)-content is a completely different pathophysiology and is still relatively unknown. That is because depleted tissue Na(+)-content is hard to detect. The complex, dominant mechanisms for the maintenance of plasma homeostasis are concealing the Na(+)-depletion in the tissues quite successfully in a stage when symptoms already may be prominent. Furthermore, we are all programmed to think about sedentary people who are rather at risk to have a salt (Na(+)) intake which is far too high. But either, competitive top athletes and engaged recreational athletes have high losses of electrolytes with sweat and might be prone to a stepwise Na(+)-depletion. All the more because they also try to have a balanced, health sodium reduced diet. One person of our research group who is used to a rather low sodium-nutrition repeatedly experienced OR-(short term-OTS)-symptoms when training loads of recreational sport activities were increasing. Getting aware about identical symptoms between OR and total body Na(+)-depletion in another professional context the decision for a self experiment was settled. Under a given training protocol changing symptoms under low sodium-nutrition were recorded. When OR-like symptoms became prominent the training loads were maintained but stringent Na(+)-substitution was performed instead of the usually recommended resting period. As experienced before, typical symptoms such as sleeping disorders, harassed feeling, high diuresis, thirst and increasing blood pressure developed within 2 weeks with the increased training loads and the usual low Na(+)-nutrition. This was before plasma sodium decreased below the physiological range. High Na(+)-substitution instead of a resting period enabled the recovery from OR symptoms within some days. Out of various articles we choose and report some interesting further medical phenomenon where our hypothesis of Na(+)-depletion as a trigger mechanism might give new ideas for identifying pathophysiological mechanisms. The hypothesis: Tissue Na(+)-depletion triggers OR- and OTS- development via the renin-angiotensin-aldosterone system which initiate at first a stimulation and then exhaustion of the sympathetic system., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Purpose: Continuous veno-venous hemofiltration (CVVH) and mixed acidemia often occur simultaneously in critically ill patients. In a previous study in non-acidemic pigs we found that colloids and CVVH interact specifically with respect to hemodynamic stability, with favorable effects for 6% HES 130/0.4 versus 4% gelatine (GEL) infusion. In a porcine model, we investigated whether these colloid-type associated differences are still dominant under acidemic conditions., Methods: We utilized 5 groups, a non-acidemic reference group receiving HES130 and CVVH; two acidemic groups receiving HES130 infusion (one with and one without CVVH); and two acidemic groups receiving GEL infusion (one with and one without CVVH). Mixed acidemia (pH ~7.20) was established by low tidal volume ventilation and acid infusion. Stable acidemia/CVVH application was maintained for 3 hours. Hemodynamics and blood gases were recorded., Results: Mixed acidemia led to a significant decrease in MAP and increase in MPAP in all groups. CVVH led to a further decrease in MAP but improved MPAP. During CVVH, HES130 ensured significantly higher MAP, Hb, and DO2 values than GEL infusion. In the groups without CVVH these differences between HES 130/0.4 and GEL were not observed., Conclusions: As in a previous study in non-acidemic pigs, we found a colloid-specific influence of HES130 versus GEL on hemodynamics during CVVH under acidemia. Again, HES130 infusion may lead to favorable effects. In contrast, acidemia without CVVH application was dominant over the impact of a respective colloid. The application of a CVVH seems to be an important trigger for the overall circulatory response to a particular colloid.

Minipigs are frequently used for scientific research as they are easy to handle and the dimensions of their vascular system do not change after 20 months of age. Although surgical interventions under anaesthesia are often performed in the supine position the effects of this positioning on lung functionality in minipigs have not been systematically described. This study aimed to analyse the influence of supine positioning on the macrostructure of the lung and the pulmonary density by the use of computed tomography imaging in pre-adult Göttingen minipigs. Twelve pre-adult female minipigs were used in the study and lung density was investigated in both the prone and the supine positions. The time between the scans in prone and supine positions was less than 5 minutes (296 ± 6 sec). In the prone position lung density did not differ between the dorsal and ventral part of the lung (-641 ± 72 Hounsfield units [HU]). However in the supine position there was a ventrodorsal gradient of decreasing density (ventral part of the lung: -497 ± 106 HU, dorsal part of the lung: -723 ± 51 HU). The changes in lung density were not accompanied by changes in lung volume (829 ± 191 ml). These results suggest an influence of the body position on the ventilation/perfusion (V(A)/Q) matching of the lung which could possibly result in lowered lung oxygenation as well as in an increased heart activity in the supine position. Additionally, due to the steep course of the vena cava caudalis from the caval foramen in the diaphragm across to the heart (in contrast to the more shallow course in the prone position) the activity of the heart necessary to pump the venous blood to the right atrium has to be higher in the supine position than in the prone position. In pigs the capacity of the heart to increase frequency is limited due to a diastolic/systolic (D/S) ratio <1. Supine positioning may possibly increase their risk of cardiovascular complications.

The most common method of controlling acidemia during lung-protective ventilation is CO₂ removal with an extracorporeal lung assist (ECLA) system. Another possibility to prevent acidemia is based on intravenous (i.v.) application of tris-hydroxymethyl-aminomethane (3 mol/L, THAM) buffer, which can bind hydrogen protons and which can be removed from the body via renal replacement therapy (RRT). We investigated whether RRT combined with predilutional (prefilter) THAM-application provides an alternative to ECLA for a rescue situation. For this, anesthetized pigs, 40 kg of body weight, six animals per group, underwent 5 h of acidemia (pH 7.19-7.24) induced by acid infusion and permissive hypercapnia (low tidal volume ventilation, PaCO₂ 80-90 mmHg). Isovolemic, high-volume hemofiltration (HVHF) was operated with predilutional THAM-infusion for treatment. To evaluate adverse effects of this approach, we set up further groups: HVHF with postdilutional (post-filter) THAM-application; i.v.-THAM without HVHF; normal pH homeostasis with HVHF. Acid-base parameters, hemodynamics, renal function, and lung morphology were investigated. HVHF with predilutional THAM-infusion of 8 mmol/kg/h allowed fast pH normalization, significant reduction in PaCO₂ to 56 mmHg and tolerable hemodynamics. HVHF alone or lower dose i.v. THAM (2 mmol/kg/h) failed to produce a comparable result. A postdilutional THAM infusion reduced hemodynamic tolerability and increased lung edema formation. HVHF in pigs with normal acid-base status resulted in a decreased base excess and urine acidification. In conclusion, predilutional THAM-application and HVHF corrected the acid-base disorder and improved pulmonary hemodynamics. Further studies are necessary to optimize the protocol including the dosage., (© 2011, Copyright the Authors. Artificial Organs © 2011, International Center for Artificial Organs and Transplantation and Wiley Periodicals, Inc.)

In recent years, acidosis has been of growing interest in intensive care medicine. Most animal models only provide a short-term investigation of the effects of acidosis. They are not suitable for research on interactions with extracorporeal organ support (here continuous venovenous hemofiltration, CVVH). The rationale for this study was to establish a porcine model of prolonged mixed acidemia, which is suitable for research on the interactions of acidemia and CVVH. After the induction of anesthesia in pigs (40 kg), acidemia was induced and maintained in one group with a bolus of 0.4 mol/L lactic acid followed by continuous infusion and a reduced respiratory frequency (lactic acid-group, n = 4). In another group, mixed acidemia was induced with a 0.4 mol/L acid solution (lactic and hydrochloric acid) and low tidal volume ventilation (mixed acidemia-group, n = 8). To get first proof of the model's suitability to operate over an extracorporeal circuit, CVVH was additionally performed in seven pigs (mixed acidemia/CVVH-group, n = 7). The target for the pH was 7.19-7.24. The targeted pH was constantly missed in the lactic acid group, whereas it was successfully maintained for 3.5 h in four out of eight pigs of the mixed acidemia group, and in five out of seven pigs of the mixed acidemia/CVVH group. The CVVH was performed successfully for 3 h in all pigs of the respective group. The mixed acidemia model was sufficient to maintain a low pH within a narrow range for some hours and enabled research on hemofilters in vivo., (© 2010, Copyright the Authors. Artificial Organs © 2010, International Center for Artificial Organs and Transplantation and Wiley Periodicals, Inc.)

Objective: In a porcine, aortic graft model we found 5 animals to develop and survive unnoticed, complete infrarenal aortic occlusion and 2 pigs with an acute occlusion but rather unspecific clinical symptoms. We investigated the pigs' vascular system to classify the anatomic capabilities to compensate for an acute abdominal aortic occlusion., Design of Study: Retrospective analysis of CT scans and clinical data to specify unexpected results in a case series of infrarenal occlusion in a porcine model., Setting: Collaborative study of experimental and clinical departments., Subjects: Fifteen healthy female minipigs., Interventions: All pigs underwent an infrarenal aortic graft intervention. Anesthesia and perioperative management of the animals were preformed along the standard operating procedures of the local Department of Experimental Medicine. All animals received perioperative antibiotics, ASS, and carprofen for postoperative analgesia. Arterial pressure, heart rate, body temperature, and diuresis were monitored during surgery and therapeutic interventions., Main Outcome Measures: Contrast media based computed tomography (CT) with total body scans were performed at 1, 4, 10, 12 weeks after surgery. Comparable scans of cardiovascular healthy subjects (humans and pigs) and patients with a Leriche's syndrome were analyzed., Results: Neither acute (within the first 12 h after surgery) nor stepwise total aortic occlusion show unmistakable clinical signs. In pigs, the epigastric artery (EGA) - which is in connection with suprarenal lumbal arteries, subclavian and external iliac artery - is highly developed associated to the high number of mammary glands of about 7 on one side. In humans, the ratio of aortic to EGA-diameter is 1 : 0.15. In minipigs we found a ratio of 1 : 0.43 which changed during aortic occlusion resulting in a ratio of 1 : 0.58. Pigs with a slowly developing occlusion demonstrated an enlargement of the ureteric artery of about 210% completing a sufficient collateral system., Conclusion: While in the human Leriche's syndrome months are needed to enlarge the EGAs for a partial collateral support of an infrarenal aortic occlusion the pig's EGA is a naturally sufficient collateral system capable to cover immediately for an acute infrarenal aortic occlusion. Further collateral enlargement even provides a permanent, sufficient hind limb perfusion in pigs. As the sufficient collateral system probably reduce pressure and shear rates in the infrarenal aortic segment after cross clamping, pigs might have a higher predisposition to produce early thrombosis related graft occlusions tan humans.

Background: C-reactive protein (CRP) is a possible causative factor of the destructive processes observed during the weeks after myocardial infarction., Methods: We developed a clinically relevant animal model including the removal of CRP from blood plasma utilizing a specific CRP adsorber and the visualization of the infarct scar in the living animal by cardiovascular magnetic resonance imaging as a tool to investigate the impact of CRP after acute myocardial infarction., Results: We describe the facets of this model system and kinetics of clinical blood parameters like CRP and troponin. In addition, we demonstrate the potency of CRP apheresis reducing CRP levels by ~70% in the established treatment system., Conclusion: We showed for the first time that it is possible to conduct apheresis at the following 2 days after acute myocardial infarction in a porcine infarction model and to analyze the infarct by cardiovascular magnetic resonance imaging at day 1 and 14., (Copyright © 2010 S. Karger AG, Basel.)

Background: Reduced haemocompatibility and early filter failure during continuous venovenous haemofiltration (CVVH) can be attributed to various aspects from filter engineering to rheological problems. Still, little is known about the impact of acidaemia and hypoxaemia on the haemocompatibility of a CVVH. In a porcine model, we investigated blood and coagulation parameters, filter performance and blockage of filter capillaries to assess the impact of acidaemia and hypoxaemia on haemocompatibility., Methods: Pigs were assigned to three groups (n = 6). One group received mixed acidaemia (pH 7.2) by acid infusion (0.2 M of lactic acid and 0.2 M HCl diluted in normal saline) and low tidal volume ventilation (6-8 mL/kg(-)(1)), one group underwent an additional hypoxaemia (pH 7.2; PaO(2) < 70 mmHg) and another was treated with normal saline and normoventilation (control group; pH 7.4). To accelerate biocompatibility reactions, CVVH was operated with reinfusion of the filtrate to the venous line for 3 h based on standardized heparinization., Results: Acidaemia led to a contradictory pattern with respect to prothrombin time (prolongation), activated partial thrombin time and activated clotting time (acceleration). In comparison to normal pH homeostasis, acidaemia led to increasing activation markers such as terminal complement complex marker sC5b-9, thrombin-anti-thrombin complexes (TAT) and D-dimers. Additional hypoxaemia intensified activation with regard to TAT and complement complex marker sC5b-9. Platelet counts suffered from acidaemia and a tendency for higher rates of blocked hollow fibres was found., Conclusion: Acidaemia led to deteriorated haemocompatibility reactions to a CVVH circuit. The coagulation pattern developed towards complications for the coagulatory state.

Purpose: Hypoxemia and acidemia (hypoxemia/acidemia) are serious complications in the critically ill and often occur in unstable patients exposed to extracorporeal organ support. Still, little is known about the biocompatibility interactions of hypoxemia/acidemia with extracorporeal circuits (ECC). Existing animal models often include the release of mediator cascades (sepsis-, lung injury models) or are based on small laboratory animals. We established a porcine model of hypoxemia/acidemia without an underlying disease and further challenged the situation with an extracorporeal circuit (ECC)., Methods: Hypoxemia/acidemia were induced (3.5 h) and maintained (3 h) in anesthetized pigs (40 kg) by a stepwise reduction in oxygenation, infusion of 0.4 mol.l⁻¹ lactic and hydrochloric acid and by low tidal volume ventilation, targeting an PaO₂ < 70 mmHg, SvO₂ < 65%, pH ~ 7.2. Venovenous hemofiltration (CVVH) operated in recirculation mode without volume exchange was chosen to prove the suitability of the model for studies on ECCs under clinical conditions (ECC group, n=6). Another 6 animals underwent the same protocol except for the CVVH (reference group, n=6)., Results: The median PaO₂ during hypoxemia/acidemia was 62 mmHg, the median SvO₂ was 38%, and the median pH was 7.22. Hypoxemia/acidemia was successfully induced and maintained for 6.5 h in all pigs. CVVH could be performed for 3 h with blood flow rates up to 300 ml.min⁻¹ and filtrate rates up to 60 ml.min⁻¹., Conclusions: Our model provides hypoxemia/acidemia with blood gas values comparable to critically ill adult patients for several hours, during which it is possible to perform CVVH. Thus, it enables research on the biocompatibility reactions of extracorporeal circuits under intensive care conditions.

The aim of this study was to find the fastest, easiest and safest method of achieving orotracheal intubation for general anaesthesia in laboratory pigs. Twenty-one Yorkshire x Landrace crossbreed male castrated pigs (32.9 +/- 4.8 kg) were investigated. Dorsal and ventral recumbency are the alternatives most frequently described for animal positioning during intubation procedures. Based on standardized induction of general anaesthesia using pentobarbital and remifentanil, the dorsoventral and ventrodorsal positions were compared with regard to the time needed, changes in oxygenation and circulatory response. Positioning was found to be crucial for fast orotracheal intubation. The time required for safe intubation is significantly shorter with the ventrodorsal position (17.3 s) in comparison with the dorsoventral position (58.4 s; P < 0.001). Hypoxia did not occur in either group. A significant drop in systolic blood pressure was observed in both groups. Diastolic and mean arterial pressures were not influenced by intubation. A significant increase in heart rate was observed in pigs intubated in ventral recumbency, but not after intubation in the dorsal position. Preoxygenation before intubation is vitally important for preventing hypoxia. With regard to clinical practice, the haemodynamic changes observed in this investigation do not appear to be relevant, as the mean arterial pressure was not altered and heart rates only increased moderately. It may be concluded that the ventrodorsal position can be recommended for orotracheal intubation in pigs as the first choice for providing a smooth and fast airway.

Control of hepatitis B virus (HBV) infection has been a continuous challenge in the management of patients with advanced kidney disease. HBV infections are still difficult to treat, so the main objective is disease prevention by timely immunization with hepatitis B surface antigen vaccines. In the general population, a simple two- or three-standard-dose vaccination strategy has been proven to be highly successful. In contrast, an intensified and more tailored approach is evolving in patients with advanced renal failure.

Introduction: Small intravascular volume, pathophysiological hemorheology, and/or low cardiac output [CO] are assumed to reduce available blood flow rates via common dual-lumen catheters (except for those with a right atrium catheter tip position) in the critically ill patient. We performed an experimental animal study to verify these assumptions., Methods: Anesthetized, ventilated pigs (35 to 50 kg) were allocated to different hemorheological conditions based on the application of different volume substitutes (that is, colloids and crystalloids, n = 6 to 7 per volume substitute). In a second step, allocation to the final study group was performed after the determination of the highest values for access flow (Qa) via an axial dual-lumen catheter (11 French, 20 cm long, side holes) placed in the femoral vein. High Qa rates (>300 ml/minute) were allocated to the dual-lumen catheter group; low Qa rates were switched to a 'dual-vein approach' using an alternative catheter (8.5-French sheath) for separate blood delivery. Hemodynamics (CO and central venous pressure [CVP]) and blood composition (blood cell counts, plasma proteins, and colloid osmotic pressure) were measured. Catheter tip positions and vessel diameters were exemplified by computed tomography., Results: Forty-four percent of the animals required an alternative vascular access due to only minimal Qa via the dual-lumen catheter. Neither hemorheologically relevant aspects nor CO and CVP correlated with the Qa achievable via the femoral vein access. Even though the catheter tip of the alternative catheter provided common iliac vein but not caval vein access, this catheter type enabled higher Qa than the dual-lumen catheter positioned in the caval vein., Conclusion: With respect to the femoral vein approach, none of the commonly assumed reasons for limited Qa via the arterial line of an axial dual-lumen catheter could be confirmed. The 8.5-French sheath, though not engineered for that purpose, performed quite well as an alternative catheter. Thus, in patients lacking right jugular vein access with tip positioning of large-French dual-lumen catheters in the right atrium, it would be of interest to obtain clinical data re-evaluating the 'dual-vein approach' with separate blood delivery via a tip-hole catheter in order to provide high-volume hemofiltration.

Background: Main factors for the overall performance of haemofilters (HF) are membrane features and filter durability without clogging/clotting of capillaries. However, the venous line resistance (Pv) is a powerful force for net filtrate flux resulting in haemoconcentration and thus is enhancing the phenomenon of membrane clogging. Therefore, we hypothesized that catheter type, as it is associated with Pv-levels, contributes to the extent in which filter longevity and filtration performance are restricted due to blocked hollow fibres., Methods: Heparinized porcine blood (5 IU/ml) was circulated in an in vitro system for haemofiltration (FH6S-filters were used, Ca. Gambro). Three different sizes of catheters for peripheral vein access (Vygonuele V, Ca. Vygon) were alternately inserted into the circuit for blood return from the filter to the reservoir. To produce Pv-levels lower than commonly induced by Shaldon catheters, a 14G-vygonuele was used. Pv-levels standard for 11-12 French catheters were provided by using a 16G-vygonuele. To produce Pv-levels common for low-French or tri-lumen catheters, a shortened 18G-vygonuele was used. The respective Pv-levels attained were compared with respect to the overall filtration performance (system pressures, haemocompatibility and sieving coefficients)., Results: Catheters of 14 and 16G enabled transiently maximal blood flow (Qb)/filtration rates (Qf) of 300/60 ml/min and continuous filtration with Qb/Qf of 200/40 ml/min. The shortened 18G catheter reduced maximal flow rates down to Qb/Qf of 200/40 ml/min, and continuous flow rates down to Qb/Qf of 125/25 ml/min. At the end, median values for blocked hollow fibres were, 35% in the 14G-group, 40% in the 18G-group and 70% in the 16G-group. Haemocompatibility appeared to be higher in the 14G-group with respect to various parameters when compared with the other groups., Conclusions: The flow resistance by the catheter chosen for haemofiltration clearly influenced the filtration performance. Thus, investigations focused on compatibility of catheter type as it related to Pv-levels with the particular method of renal replacement therapy that should be performed. This point could be crucial in reducing filter clogging and haemostasis during CVVH.

The effects of surgical procedures on intestinal motility are still under debate. In order to quantify the effects of duodenotomy on duodenal motility, the present study used the electric impedance technique (IMP). Ten pigs (32-40 kg) were instrumented under general anaesthesia with a central venous catheter (CVC) and a percutaneous enterogastrostomy (PEG). Duodenal phases I-III and the duration of the migrating motor complex (MMC) were measured by an IMP catheter, which was introduced non-invasively into the proximal duodenum via the PEG by endoscopy, or through surgical placement in an invasive manner via duodenotomy. There were no significant differences in the length of interdigestive phases I-III between the groups. In tendency, pigs after laparotomy and duodenotomy demonstrated slightly prolonged median duration of phase I (P = 0.1) and phase III (P = 0.7), while phase II (P = 0.1) and the MMC cycle length (P = 0.6) seemed to be shortened as compared to pigs of the PEG group. In conclusion, simple laparotomy with duodenotomy did not affect the proximal duodenal motility activity measured by the impedance technique.

Unlabelled: Both fluid management and renal replacement therapies play a fundamental role in the treatment of critically ill patients. In a recent in vitro study, we have shown specific interactions of different colloids and the hemocompatibility of hemofilters. The present study was performed to compare the five most common fluids for volume resuscitation, i.e., normal saline (SAL), hydroxyethyl starch 130 kd/0.4 (HES130), hydroxyethyl starch 200 kd/0.5 (HES200), albumin (ALB), and gelatin (GEL) with respect to their interaction with continuous venovenous hemofiltration (CVVH) in anesthetized domestic pigs., Methods: Animals (n = 63) were allocated randomly to the fluid type and the respective subgroups, which were divided into control and CVVH groups (n = 6 ndash; 7 per group). Bolus infusion of group specific fluid was followed by a bolus of heparin and the initiation of hemofiltration in CVVH groups. Thereafter, fluids were infused at constant rates, and heparin application was adjusted to keep the activated clotting time at 200 to 250 s. Hemodynamics, airway pressures, pulmonary gas exchange, diuresis, creatinine clearance, and blood cell counts were investigated during the entire procedure (10 ndash; 12 hours)., Results: Basics of in vivo effects of SAL, HES130, and ALB were not altered during CVVH. HES130 and ALB enabled stable hemocompatibility, diuresis, and hemodynamics in the respective groups. In contrast, organ functions were significantly different between control and CVVH groups when animals were treated with GEL or HES200. In particular, during CVVH, HES200 led to reduced platelet counts, deteriorated hemodynamics, and increasing airway pressures during CVVH. GEL led to increasing airway pressures, a decrease in pulmonary gas exchange, deteriorated hemodynamics, altered renal histomorphology, reduced platelet counts, and reduced hemoglobin., Conclusions: Direct in vivo effects of colloids in anaesthetized and ventilated pigs are not predictable for their effects during CVVH. Interaction between CVVH and every volume substitute occur in a highly specific manner. This observation could be helpful to explain contradictory study results and should be considered for future study designs.

Flow distributions are critical determinants in the function of hemofilters. Despite their importance, however, flow distributions cannot currently be measured in filters during experimental or clinical applications. Here, we demonstrate that the thermal conduction properties of extracorporeal circuits may provide a tool to overcome this limitation. More specifically, we show that thermography provides an indirect approach to visualize differences in regional perfusion rates through temperature profiles on the filter surface. Thermograms were recorded using a TVS700 system (Ca. Goratec) during recirculating in vitro hemofiltration of porcine blood. Different test protocols were executed to characterize the contribution of thermal conduction and convection to the measurable changes in the temperature at the surface of the filter housing. For comparison and validation, these experiments were supplemented by computer tomography (CT) of filters after dye injection. Thermography enabled real-time visualization of the flow distributions in a hemofilter. Moreover, 'point' trends taken from different regions of the filter provided quantitative information about changes of flow distributions in response to changing experimental conditions. Our preliminary data suggest that thermography is a promising new approach for assessing the principles and time-related changes in flow distributions in hemofiltration. As expected, resolution is lower than that in CT measurements and further studies will be necessary to determine the smallest temperature gradient that still identifies differences in regional perfusion rates. Given its potential to develop into an inexpensive tool for the 'bedside' level monitoring of flow distributions during clinical studies, further investigation of thermography is highly desirable.

To investigate the effects of propofol and fentanyl on the postprandial duodenal motility the intraluminal impedance technique was used. Six pigs were instrumented with a central venous catheter, a percutaneous enterogastrostomy (PEG), and an impedance catheter, which was introduced via the PEG into the duodenum through endoscopy. Over the following 3 d, duodenal motility was measured for 8-hour periods. Measurements were taken on each subject under 3 different sets of conditions: in the conscious unrestrained pig, during propofol sedation, and during sedation with propofol-fentanyl. Both, after morning feeding and during gastric nutrition via the PEG, duodenal feeding patterns and duodenal phase II of the migrating motor cycle were shortened during propofol and propofol-fentanyl sedation. In contrast, the duration of phase I was prolonged by propofol and propofol-fentanyl. In conclusion, either propofol or propofolfentanyl sedation shortens duodenal feeding patterns, as well as phase II of the migrating motor cycle.

Background: Apart from their standard applications, haemofiltration (HF) and plasmafiltration (PF) may provide helpful therapy for sepsis, multiple organ- and acute liver-failure. Some colloids cause either decreases or increases in blood cell agglomeration. We hypothesized that solutions which reduce cell aggregability may lead to both improved filter clearance and better haemocompatibility due to decreasing rates of clogged hollow fibres., Methods: Heparinized porcine blood (5 IU/ml) was used in an in vitro circuit. The filter types tested were from GAMBRO: HF66D (effective membrane surface: 0.6 m2) and PF1000N (effective membrane surface: 0.15 m2). Albumin (ALB), hydroxyethyl starch (HES) 200/0.5, HES 130/0.4, gelatin (GEL) or normal saline (0.9%) were added to the blood (n = 6/group). Recirculation systems were run for 2 h. Spontaneous haemolysis and filter resistance >420 mmHg were selected as indications of maximal flow rates. Sieving coefficients were determined for 17 parameters at the lowest and highest blood flows and filtration rate., Results: Based on the filter types used, supplementation of ALB and HES130/0.4 led to an improved filter clearance without increasing the number of clogged capillary membranes or causing impaired haemocompatibility. Sieving coefficients for most solutes were independent of volume substitute and flow rate. Haemocompatibility and filter clearance deteriorated after addition of HES200 or GEL to the blood., Conclusions: Under standardized in vitro conditions, we found that colloids which reduce cell aggregability cause improved HF- and PF-performance. This phenomenon may provide new options for higher clearances and may lead to new concepts in low dose anticoagulation.

During extracorporeal circulation, activation of the complement system as a bioincompatibility reaction upon contact with artificial surfaces is well known. This may induce adverse effects for the patient as well as for the bioreactor when using a hybrid liver support system. At present, the achievable clearance rates in hybrid liver support systems are restricted by the limitations of blood flow through the capillaries of the plasma filter. In order to enhance clearance rates it is desirable to develop a high-flow plasma separation unit in which the activation of the complement system is as low as possible. Conventional plasma filters operating in normal and in inverse (blood flow around the capillaries and the capillary lumen serving as plasma compartment) modes were investigated applying an in vitro circuit (n = 4 in each group). For this purpose heparinized porcine blood was used. Complement activation was measured as generation of the soluble membrane attack complex, SC5b-9. With a normal flow mode the maximum achievable filtration rates varied from 25 to 40 mL/min, whereas the maximal filtration rates with an inverse flow mode varied from 40 to 100 mL/min. While these values were significantly higher than those with regularly operating plasma filters, there was no significant difference between normal and inverse mode in the production of SC5b-9. We conclude that the outer surface of the investigated conventional plasma filtration membranes is suitable for an inverse mode and does not lead to an excessive activation of the complement system upon dynamic blood contact. Thus, the inversion of the blood and the plasma compartments may be a possible way forward in the development of a high-flow plasma separation filter.

In order to investigate the effects of a single dose of ketamine on duodenal motility, the present study focused on the electric impedance technique. Five pigs (32 to 40 kg, CVC group) were instrumented with a central venous catheter 1 d before measurements. The next day, general anesthesia was started and maintained via central venous catheter by propofol and fentanyl. In contrast, the pigs of the KETA group (n = 5) received ketamine intramuscularly prior to the induction of anesthesia by the injection of propofol-fentanyl via an ear vein. An intraluminal impedance catheter was manually introduced into the proximal duodenum. Measurements were recorded for 4 h. The KETA group showed a median duration of phase II that was shortened by 35%, while phase I was prolonged by 73% (P < 0.05). In conclusion, when gastrointestinal motility has to be investigated, the effects of a single dose of ketamine, even for premedication, should be taken into consideration.

Background: In contrast to humans, young pigs naturally have a low COPpl (12-16 mmHg versus 22-26 mmHg in young humans). Thus, behavior occurring when volume management is performed similar to human medicine might be different. Potentially underestimated intra- to extravascular fluid and solute shifts could influence time course of variables investigated. That is why we studied whether differences in the basic protocol for infusion therapy and different levels of COP according to age or body weight, respectively - impair reproducibility and reliability of experimental results., Methods: Group A [n=6, lower body weight (LBW < 22 kg)] was treated with unrestricted infusion rates (UIR) adjusted to maintain a constant blood pressure; group B [n=6, higher body weight (HBW > 28 kg)] was treated with a restricted continuous infusion rate for fluid balance (RIR); group C (n=6) combined HBW and UIR protocol. Blood pressure, plasma solutes, diuresis, and peritoneal fluid were analysed. Statistical analysis was performed using Kruskal-Wallis test and Wilcoxon test., Results: UIR-treated pigs with LBW (group A) developed ascites and demonstrated time dependent decreases of plasma solute concentrations whereas in pigs of group C mainly diuresis was increased and subcutaneous edema occurred. None of the protocols enabled constant blood pressure., Discussion: In young pigs (LBW), an adaptive volume substitution using crystalloids to standardize blood pressure may induce fluid extravasation; in turn data may not show statistical significance, stable hemodynamics may not be achieved and changes in plasma solute concentrations may lead to false interpretations. RIR-strategy provided reproducible, plausible results and thus should be recommended in combination with pigs >29 kg BW for the use of porcine models.

Capillary membrane filters are devices commonly used for plasmapheresis and recirculating detoxification systems, including plasmaseparation units. Besides clinical trials and case reports, research on plasmafiltration techniques is carried out using in vitro systems. Notably, such hemoperfused in vitro systems require anticoagulation protocols suitable for investigating the clearance performance and hemocompatibility of a plasmafilter. This study analyzes how different heparin concentrations affect filtration performance and the hemocompatibility under conditions typically employed during in vitro experiments. Porcine blood from healthy slaughtered animals was used in an in vitro circuit. The blood collecting system was primed with heparin. Depending on the study group, blood was anticoagulated with either 2.5 IU/ml or 5 IU/ml heparin. The filters were n=10 PF1000N (effective membrane surface: 0.15 m2). For each experiment, the maximal permissible flow rate was established by determining the point where spontaneous hemolysis occurred and/or pressure values exceeded the highest limit of the pressure units on the apheresis monitor. Sieving coefficients of ten parameters, platelet counts, and coagulation patterns were determined at the lowest and highest blood flow (Qb) and filtration rates (Qf). Except for the activated clotting time (ACT), both anticoagulation protocols caused the blood to respond to its exposition to the plasmafilters concerning the activation and inhibition of the coagulation system. A significant decrease in platelet counts did not occur. There were sporadic differences in sieving properties between the groups. However, heparinization with 2.5 IU/ml resulted in unreproducible flow rate profiles, and pressure levels were higher than in the group with 5 IU/ml heparin. Compared to heparinization at 2.5 IU/ml, higher levels of heparinization stabilize rheological properties of the blood, and thereby increase the reliability of data obtained from plasmafiltration experiments carried out in vitro.

Animal models have become an essential tool in the investigations of gut motility under experimental conditions. To determine the influence of various anaesthetic drugs on the motility pattern of the gastroduodenal tract, a new long-term model has had to be developed for allowing measurements in conscious and unrestrained as well as in sedated and analgosedated pigs. Since mechanical ventilation influences gut motility, it was necessary that this animal model enabled the investigation of the effect of drugs causing sedation and analgosedation during spontaneous breathing. Seven male, castrated pigs, German landrace, 32-40 kg bodyweight (BW) were investigated in this study. After habituation of the pigs to local housing conditions over 5 days, the animals were trained over 4 days to prepare for experimental situations and investigators. Pigs were inserted with a central venous catheter and with percutaneous enterogastrostomy (PEG) under general anaesthesia. Intestinal motility was measured by intraluminal impedancometry. The catheter was introduced over the PEG into the stomach and positioned into the duodenum by duodenoscopy. Measurements were done in conscious, unrestrained pigs and with sedated, and analgosedated animals on subsequent days. The habituation and training of the pigs to the investigators and for the laboratory conditions took between 7 and 9 days. The initial anaesthesia protocol for the instrumentation using remifentanil/propofol led to pyloric spasm and was thus unsuitable for duodenal intubation with an endoscope. In contrast, a combination of ketamine/propofol enabled this procedure. It was practicable to measure gut motility in conscious, unrestrained pigs. Spontaneous breathing was sufficient under propofol sedation and analgosedation using fentanyl-propofol. Systematically local application of polividon iodine in the area of the subcutaneous catheters avoided the necessity of using systemic prophylactic antibiotics. In conclusion, the habituation and training for 9 days enabled the measurement of gut motility by intraluminal impedancometry in conscious pigs. The insertion of the catheter was done during general anaesthesia using a combination of propofol and ketamine. For the future determination of gut motility performed under general anaesthesia, each sedation and analgosedation concept has to be evaluated to see whether it allows spontaneous breathing or whether mechanical ventilation is necessary.

Plasmaseparation is a treatment under discussion for critically ill patients, especially in sepsis and multiorgan failure. These patients receive a variety of different fluid substitutes, including hydroxyethylstarch (HES). HES is known to influence rheological properties, but nothing is known about the possible interactions between HES and the plamaseparation procedure. We used an in vitro plasmaseparation circuit with heparinized porcine blood. Before priming the system, 2 liters of blood were supplemented by adding 100 ml of either NaCl 0.9% or HES (n=6 in each group). We monitored the transmembrane (TMP) and the filtration pressure (PF) and measured free plasma hemoglobin (free Hb) and platelet counts before and after the two hours plasmaseparation procedure. The final transmembrane pressure was significantly higher with HES substitution. In the HES group we found negative filtration pressures from the very beginning with a significant further decrease toward the end of the experiments. A significant increase in free Hb and decrease in platelet counts were noted only in the HES group. Volume substitution with HES leads to impaired filtration properties and deteriorated hemocompatibility in in vitro plasmaseparation. Further studies have yet to evaluate whether or not the effects described also occur under clinical conditions.

In Europe, capillary membrane filters are the favored method for plasmapheresis and detoxification systems including a plasma separation unit. Using capillary membrane filters clearance depends on the filtration rate and the sieving coefficient (Qs) for the plasma substrates. We investigated whether the increase of the effective membrane surface of plasmafilters enables higher clearance rates than a small plasmafilter. Heparinized porcine blood (5 I.U./l) of healthy pigs was used for an in vitro circuit. Filter types used were from GAMBRO: n = 5 PF1000N (effective membrane surface: 0.15 m(2)), n = 4 PF2000N (0.35) and from ASAHI: n = 4 PLASMAFLO OP-05W (L) (0.5). Spontaneous hemolysis was chosen to indicate the individual threshold limit for maximal flow rates. Sieving coefficient was determined for ten parameters at lowest and highest blood flow (Qb) and filtration rates (Qf). PF1000N enabled maximal flow rates as high as PF2000N and PLASMAFLO (Qb/Qf in ml/min: 200/40). Qs for most substrates were similar between the filter types and the flow rates. Compared to PLASMAFLO, the filter type with the largest membrane surface, the PF1000N provided significantly better Qs for triglycerides and albumin and a trend for a better elimination of cholesterol, GGT, and LDH. The sieving properties of the PF2000N ranged between PF1000N and PLASMAFLO. Under standardized in vitro conditions, the larger plasmafilters tested did not improve the overall performance of the plasmapheresis procedure. Thus, enlargement of the membrane surface such as provided by two commercially available and clinically well-established plasmafilters could not be proven to enable higher clearance rates., (Copyright 2002 Wiley-Liss, Inc.)

As the low clearance rate of plasmaseparation limits its use in the treatment of patients suffering from liver failure, sepsis or MOF, we intend to develop strategies for a plasmaseparation unit which increases plasmafiltration rates. Our first question focused on whether commercially available plasmaseparation filters, and in particular their membranes, are suitable for the inversion of blood and plasma compartments. This experimental study was performed using in vitro systems. Commercially available plasmafilters PF2000N (Gambro) and Plasmaflo (Asahi) were compared in both their normal operating mode with blood flow through the capillary lumen, and in the inverse mode. Inverse mode means that blood flows through the outer space of the capillaries while plasma was obtained from the lumen. Heparinised porcine blood (5 I.U./ml) was used in a heated, recirculating in vitro circuit. Our main results were that the normal use of both filter types Plasmaflo and PF2000N enabled maximal blood flows (Qb) of 200 ml/min and filtration rates (Qf) of 25-40 ml/min. Operating the filters in the inverse mode enabled Qb up to 500 ml/min and Qf up to 100 ml/min. Hemolysis, platelet counts and coagulation parameters did not differ significantly regardless of whether the normal or inverse mode was used. The tested plasmafiltration membranes appear to be suitable for use in inverse mode. Although in our experiments, hemocompatibility tests did not indicate severe problems induced by the module geometry, the development of a module specially constructed for blood flow outside of the hollow fibers appears to be necessary in order to minimise shunts and low perfusion areas.

Commonly used materials incorporated into dynamic culture systems typically show the feature of adsorption of lipophilic xenobiotics. Yet, this phenomenon is strongly limiting the use of dynamic culture models and ex vivo organ perfusions in pharmacological and toxicological research. The aim of the study was to characterize different materials with respect to their capacity for drug adsorption and to find methods or materials to reduce the loss of substrate by adsorption in order to improve the use of dynamic in vitro systems. The adsorption of different xenobiotics (lidocaine, midazolam, lormetazepam, phenobarbital, testosterone, ethoxyresoroufine) to tubes used in dynamic in vitro systems (polyvinyl-chloride, silicone) were investigated and compared to a new material (silicone-caoutchouc-mixture). In addition, the role of protein deposition onto the tubing was studied and it was investigated whether it was possible to reach saturation of the inner tube surface by pre-loading it with the test compound. We found that silicone tubes provided the highest comfort with respect to handling and reusability, but they also demonstrated the highest capacity for substrate adsorption. Polyvinyl-chloride was the second best in handling but also demonstrated a high complexity in its adsorption behavior. The silicone-caoutchouc-mixture reached acceptable experimental results with respect to its handling and demonstrated a very low capacity for substrate adsorption.

Background: Plasmapheresis has been used for the treatment of acute liver failure (ALF). In these patients, hypoalbuminemia is often observed. Since albumin improves the disaggregability of erythrocytes, hypoalbuminemia might deteriorate rheology and thus influence the overall performance of plasmapheresis., Methods: Hypoalbuminemia was mimicked by using porcine blood because of its physiologically low albumin/globulin ratio (AGR). Filters (n = 16) were integrated in a closed extracorporeal in vitro system. In the control group (n = 8), native porcine blood (AGR 0.8) was used. In the study group (n = 8), we used porcine blood supplemented with human albumin to obtain the human AGR value of 1.2. Two different heparinization protocols were compared in each group (2.5 IU/ml: n = 4 with albumin and n = 4 without albumin versus 5 IU/ml: n = 4 with and n = 4 without albumin)., Results: In both heparinization protocols the higher AGR led to lower transmembrane pressure (TMP) levels compared to the lower AGR. The reduced TMPs enabled higher blood flow and filtration rates., Conclusion: Maintenance of a physiological AGR in ALF patients might improve the performance of plasmapheresis and--as opposed to raised heparinization--contribute to a safer application., (Copyright 2001 S. Karger AG, Basel)

Pigs are widely used as models for a variety of human diseases, because many of their physiological functions closely resemble those of humans. However, information on instrumentation techniques is still scarce. In particular, experiments in conscious pigs focused on extracorporeal circuits are connected to a variety of methodical problems with respect to the handling of the animals. Usually, pigs are placed in restraint-slings during the application of an extracorporeal system. However, this method of restraint may lead to excessive mental distress even in trained animals. The latter might influence the results and certainly affects principles of animal welfare. Our own experiences with instrumented, conscious, but unrestrained dogs encouraged us to modify methods used for the fixation of in-dwelling central venous catheters in dogs with special regard to the species specific behaviour and phenotype of pigs. A cord retractable leash (CRL) was used for maintaining a safe distance between the animal and the outer ends of the catheters. To prevent dehiscences of the required fixation sutures a new catheter bag (CB) was designed to counteract tension forces caused by the CRL's spring-mechanism. The combination of both the CRL and CB enabled us to conduct safe experiments with conscious, unrestrained pigs. We alleviated the mental distress these animals were exposed to in comparison to former methods based on restraint of the animals.

Culture media are frequently used in the evaluation of metabolical functions of hepatocytes in hybrid liver support systems (hLSS). However, media compositions differ substantially from those of plasma. Therefore, our study was designed to investigate whether current in vitro studies with medium are suitable to assess the metabolical competence of hLSS-cultures during clinical application as well as to explore whether the cell nutrition with medium provides a suitable modus operandi for stand by cultivation. Paired bioreactor cultures were perfused with either Williams' Medium E (MPB) or human plasma (PPB). About 6x108 primary pig hepatocytes (>97% viability) were cultured in three laboratory scale bioreactors designed according to Gerlach's bioreactor-concept. Different perfusion protocols were initiated after a standardised period allowing for cell attachment and reorganisation in aggregates. Whereas patterns of enzyme release were similar in both protocols the metabolical behaviour was different between MPB (anabolic state) and PPB (catabolic state). Furthermore, compared to MPB the lidocaine-MEGX-tests for PPB demonstrated lower MEGX-concentrations and a different reaction pattern. We conclude that the nutrition of hepatocytes with medium during the stand by period itself might influence the cell function and subsequently the efficacy of the hLSS-treatment during clinical application.