Background & aims: Liver surgery usually involves ischemia and reperfusion (I/R) which results in oxidative stress and cell damage. The administration of antioxidants should diminish or prevent this damage. The purpose of this study was to investigate the effect of the antioxidant vitamin E on I/R injury.Methods: We carried out a placebo-control led double-blind study on 68 patients undergoing elective, tumor-related, partial liver resection. 47 patients were qualified for the per protocol population based evaluation. The patients were randomly assigned to two groups. The day before surgery one group received three infusions containing vitamin E (600 IU = 540 mg vitamin E emulsion). The other group received three infusions of placebo.Results: Length of stay in the intensive care unit (ICU) was significantly shorter in the verum group than in the placebo group (P<0.05). There were signs of improvement for AUC AST (P<0.05), ALT and GLDH in the verum group after surgery. Serum vitamin E concentration increased after administration of vitamin E infusion and declined in both treatment groups after surgery (P<0.01). In the verum group vitamin E deficiency was prevented while vitamin E concentration remained low in the placebo group (P<0.01).Conclusions: The findings from this study indicate that preoperative administration of vitamin E is safe and that this treatment may have beneficial effects by reducing the impact of I/R injury in liver surgery. (C) 2004 Elsevier Ltd. All rights reserved.

Liver xenografts transplanted from guinea pig to rat suffer from inadequate organ reperfusion and initial dysfunction, despite sufficient complement depletion using cobra venom factor (CVF). Reperfusion injury is prevented when complement depleted donors are treated with the prostacyclin analog epoprostenol. Histological analysis suggests that epoprostenol preconditioning prevents post-reperfusion spasms of the intrahepatic branches of the portal vein and strongly reduces appearance of hepatocyte apoptosis shortly after transplantation. Cobra-venom-treated rats show breakdown of glucose metabolism and die in acute hypoglycaemia, whereas the additional application of epoprostenol restores gluconeogenesis. Consequently, recipient survival after epoprostenol and CVF treatment is significantly improved compared with animals receiving CVF only (5.1 +/- 2.6 h vs. 17.9 +/- 5.1 h). These data demonstrate that initial dysfunction of discordant liver grafts in the guinea-pig-to-rat species combination, can be overcome by the application of epoprostenol combined with CVF. Using this pharmacologic regimen, the discordant guinea-pig-to-rat model appears useful to study further questions concerning functional and immunological compatibility of a discordant liver xenograft.

Heart failure with a preserved ejection fraction (HFpEF) and heart failure with a reduced ejection fraction (HFrEF) have distinctive pathophysiologies, and thus, therapeutic approaches to the 2 disorders should differ. Neurohormonal activation drives the progression of HFrEF, and neurohormonal antagonists are highly effective in HFrEF, but not in HFpEF. Conversely, a broad range of chronic systemic inflammatory or metabolic disorders cause an expansion and inflammation of epicardial adipose tissue; the secretion of adipocytokines may lead to microvascular dysfunction and fibrosis of the underlying myocardium, which (if the left atrium is affected) may lead to atrial fibrillation (AF) and (if the left ventricle is affected) may lead to HFpEF. Anti-inflammatory drugs (such as statins and anticytokine agents) can ameliorate epicardial adipose tissue dysfunction. Statins appear to ameliorate the development of atrial myopathy (both experimentally and clinically), and in randomized controlled trials, they reduce the incidence of new-onset and recurrent AF and decrease the risk of heart failure with the features of HFpEF; yet, they have no benefits in HFrEF. Similarly, anticytokine agents appear to prevent heart failure in patients with or prone to HFpEF, but adversely affect HFrEF. Several antihyperglycemic agents also reduce epicardial fat mass and inflammation, but this benefit may be offset by additional actions to cause sodium retention and neurohormonal activation. Thiazolidinediones have favorable effects on experimental AF and HFpEF, but their antinatriuretic actions negate these benefits, and they worsen the clinical course of HFrEF. Glucagon-like peptide-1 receptor agonists also ameliorate AF and HFpEF in laboratory models, but their positive inotropic and chronotropic effects may be deleterious in HFrEF. By contrast, metformin and sodium-glucose cotransporter 2 inhibitors alleviate epicardial adipose tissue dysfunction and may reduce the risk of AF and HFpEF; yet, they may have additional actions to promote cardiomyocyte survival that are useful in HFrEF. The concordance of the benefits of anti-inflammatory and antihyperglycemic drugs on AF and HFpEF (but not on HFrEF) supports the paradigm that epicardial adipose tissue is a central pathogenetic mechanism and therapeutic target for both AF and HFpEF in patients with chronic systemic inflammatory or metabolic diseases.

In einer tierexperimentellen Arbeit wurde das Auftreten von Apoptose beim Ischämie-/Reperfusionsschaden nach Lebertransplantation untersucht. Rattenlebern wurden nach hypothermer Organkonservierung in HTK Lösung syngen transplantiert und anschliefíend das Ausmaß der Apoptose bestimmt. Dabei konnte gezeigt werden, daß es nach hypothermer Ischämie und Reperfusion zu einer apoptotischen Schädigung vor allem endothelialer Zellen kommt. Nach Vorbehandlung mit α-Tocopherol konnte die Apoptose endothelialer Zellen signifìkant gehemmt werden. Wir vermuten, daß die apoptotische Endothelzellschädigung in der frühen Reperfusionsphase eine wichtige Rolle in der Pathogenese des Ischämie-/Reperfusionsschadens nach Lebertransplantation spielt.

Since organ transplantation became a standard procedure in medicine, some interdisciplinary discussion has evolved around the availability of organs for transplantation. The shortage of available donors leads to numerous deaths on waiting lists where heart, lung and liver disease is concerned. Patients on dialysis spend years waiting for a suitable cadaveric graft. The shortage of organs has widened not only the selection criteria for cadaveric donors and the optimization of procurement but also has led to the increased acceptance of relatives and friends as living donors for kidneys, parts of the liver and maybe in the future of the lung. It has to be decided in which direction one wants to influence the discussion about the retrieval of an adequate number of organs for our waiting patients.

Xenogeneic liver transplantation in the discordant guinea pig (gp) to rat model results in hyperacute rejection within a few minutes, which is due to activation of the complement system. Currently no antibodies against gp complement factors are available, which allow activation of the gp complement system in serum or complement deposition in tissue to be detected. To close this gap, we started developing single chain Fvs (scFvs) against gpC5 and gpC5a. We generated a combinatorial library of scFv antibodies comprising the variable heavy and light chain repertoire from mice immunized with gpC5. Out of this library we selected several antibodies against gpC5 and C5a after four and six rounds of biopanning, respectively. Selected gpC5-specific scFvs were purified by metal affinity chromatography followed by size exclusion chromatography or by affinity chromatography using Protein L. Purified scFvs were able to inhibit gp complement system in a hemolytic assay and to detect gpC5 deposition in tissue. A surface plasmon resonance based assay on BIAcore was established, with which the C5 concentration in gp serum was determined to 240 microg/ml. As at least 0.04% of the normal gpC5 concentration can be detected, the test provides a powerful tool to investigate the development and the consequence of a hybrid complement system after liver xenotransplantation from gp to rat.

Im Rahmen einer Beobachtungs-Kohortenstudie wurden 138 Patienten vor und nach Nierentransplantation und 118 Patienten vor und nach Lebertransplantation zu ihrer Lebensqualität befragt. Erhebungsinstrumente waren das Nottingham Health Profile (NHP), eine Activities of Daily Life (ADL) Skala, die Subskala, “Somatisierung” der Symptom Check List (SCL-90-R), die Center of Epidemiology and Statistics Depression Scale (CES-D) zur Selbsteinschätzung sowie der Spitzer-und der Karnofsky-Index zur Fremdbewertung. Abgesehen von einer Beschwerdezunahme in einigen Bereichen während der postoperativen Frühphase zeigten sich in beiden Gruppen signifikante und anhaltende Verbesserungen der Lebensqualität schon drei Monate nach der Transplantation. Patienten nach Nierentransplantation wiesen z. T. Werte unterhalb des Beschwerdeniveaus einer Bevölkerungsstichprobe auf. Die Untersuchung zeigt, dass eine systematische Erfassung der subjektiven Wahrnehmung des Patienten auch in extremen Behandlungssituationen möglich ist und einer differenzierten Bewertung medizinischer Massnahmen dienen kann. Sie liefert damit einen Beitrag zur Entwicklung der modernen Evaluationsforschung im Sinne einer auf empirischer Evidenz beruhenden medizinischen Praxis.

Die Effektivität einer Behandlung in klinischen Studien anhand biomedizinischer Kriterien wie Überlebens- und Funktionsraten zu belegen, galt bislang als ausreichend, um ihre Einführung und Verbreitung im klinischen Alltag zu begründen. Dabei wurde oft vernachlässigt, daß sich die unter Studienbedingungen erreichten Ergebnisse in einer breiten Anwendung nicht unbedingt reproduzieren lassen - im angloamerikanischen Schrifttum wird diese Differenz durch die Unterscheidung von efficacy und effectiveness berücksichtigt.

VON HÖRSTEN, S. M. S. EXTON, J. VÖGE, J. WESTERMANN, M. SCHULT, E. NAGEL, R. E. SCHMIDTAND M. SCHEDLOWSKI. Cyclosporine A affects open field behavior in DA rats. PHARMACOL BIOCHEM BEHAV 60(1) 71–76, 1998.—Since the introduction of Cyclosporine A (CsA) for immunosuppression in solid-organ transplantation, the rate of allograft rejection has decreased substantially. However, treatment with CsA induces neuropsychological complications in patients, including affective disorders such as anxiety, disorientation, depression, aggression, paranoia, and apathy. These CsA-induced affective side effects cannot be extensively studied in humans. Therefore, this study investigates the effects of intraperitoneal CsA (20 mg/kg) injections on the open-field behavior of male Dark Agouti (DA) rats 1, 6, 12, and 23 h after drug administration on 3 consecutive days. CsA induced an increase in emotionality in DA rats 6 h after injection, reflected by decreased ambulatory activity in the open field and increased defecation. In addition, a decrease in rearing activity was observed 12 h after CsA administration. These behavioral alterations are discussed in the view of changes in cytokine profiles induced by CsA.

Electron microscopy of dimeric and trimeric single chain antibody Fv fragments (scFvs) complexed with anti-idiotype Fab fragments was used to reveal the orientation of antigen binding sites. This is the first structural analysis that discloses the multivalent binding orientation of scFv trimers (triabodies). Three different scFv molecules were used for the imaging analysis; NC10 scFv-5 and scFv-0, with five- and zero-residue linkers respectively between the VH and VL domains, were complexed with 3-2G12 anti-idiotype Fab fragments and 11-1G10 scFv-0 was complexed with NC41 anti-idiotype Fab fragments. The scFv-5 molecules formed bivalent dimers (diabodies) and the zero-linker scFv-0 molecules formed trivalent trimers (triabodies). The images of the NC10 diabody-Fab complex appear as boomerangs, not as a linear molecule, with a variable angle between the two Fab arms and the triabody-Fab complexes appear as tripods.

Immunosuppression induced by Cyclosporine A (CsA) can be behaviorally conditioned. It is unknown, however, whether a taste aversion paradigm using CsA as an unconditioned stimulus (UCS) induces alterations of blood leukocyte numbers and function. Results obtained by three-colour flow cytometry and granulocyte chemiluminescence response demonstrate that in conditioned rats, absolute numbers of lymphocyte subsets, including B, CD8+ T cells and CD4+ naive and memory T cells, and granulocyte numbers and function were significantly decreased. In contrast to the conditioned response, CsA treatment alone increased lymphocyte numbers and did not affect granulocyte function. Thus, our data demonstrate that behaviorally conditioned CsA effects can be monitored in the blood. In addition, results indicate that the CNS mediates the behaviorally conditioned immunosuppression by reducing the availability and function of granulocytes and lymphocytes.

The classical conditioning of immune parameters is commonly conducted within a conditioned taste aversion (CTA) paradigm. In this study, the immunosuppressive drug cyclosporine A (CsA) was investigated for its ability to produce both taste aversion to a novel stimulus and conditioned alterations in immune functioning. The paradigm comprised the pairing of a 0.2% saccharin solution (the conditioned stimulus; CS) with an intraperitoneal injection of 20 mg/kg CsA (the unconditioned stimulus; UCS). Upon saccharin re-presentation, a marked reduction in fluid consumption was observed, indicating aversion to the novel substance (=CTA). By using a single CsA/saccharin pairing the CTA lasted for one CS representation. However, by implementing three pairings, this effect could be extended for up to seven representations. No noticeable difference was recorded by adjusting the saccharin representation from every consecutive day to every second day. The most effective paradigm in creating CTA was subsequently investigated for its effectiveness in producing conditioned immune alterations. Animals were killed on the day of the third CS re-presentation, and immune functions assessed. Conditioned animals displayed a significant reduction in thymus and spleen weights. Effects on the spleen were further investigated, revealing a significantly reduced proliferative ability of isolated splenocytes to concanavalin A. These results demonstrate that the physiological effects produced by CsA are sufficiently salient to elicit CTA. Furthermore, the reduction in lymphoid organ weight and splenocyte proliferation induced by CsA are also conditionable using this paradigm. (C) 1998 Elsevier Science Inc.

Even though knowledge and systematic application of placebo responses in the immune system are sparse, this topic is of particular importance since it may aim at drug-dose reduction while maintaining therapeutic efficacy of treatment in clinical settings. Placebo responses in the immune system are inducible by associated learning paradigms, such as behaviorally conditioned immunosuppression. One established learning paradigm in both rats and humans is conditioned taste avoidance (CTA), where a novel taste as conditioned stimulus (CS) is paired with the administration of the immunosuppressive drug cyclosporine A (CsA) as unconditioned stimulus. By representing the CS alone at a later time point, the conditioned response is reflected by avoidance behavior toward the taste (CTA). Simultaneously, diminished cytokine production and proliferative capacity of T cells are observed, closely mimicking the pharmacological effects of CsA. This chapter provides an overview on placebo responses in the immune system and delineates actual approaches, translational aspects, and limitations of learning paradigms in clinical settings.

Bi-directional interactions between the central nervous system (CNS) and immune system are demonstrated by the modification of immune function using behavioral conditioning. However, the mechanisms by which the CNS achieves conditioned immunomodulation are still in question. Here, we report that the immunosuppressive effects of cyclosporine A (CsA) can be behaviorally conditioned in rats using saccharin as a gustatory conditioned stimulus. The conditioned effects were compared to control groups that received CsA paired with water (sham-conditioned), CsA injection on test days (CsA-treated), and unhandled rats (untreated). In conditioned animals, the mitogen-induced lymphocyte proliferation in the spleen is significantly suppressed, and the survival time of heterotopic heart allografts prolonged. These effects are paralleled by conditioned inhibition of IL-2 and IFN-γ synthesis by splenocytes. Furthermore, the CNS-induced immunosuppression is mediated neuronally and not via the blood, since the conditioned reduction of proliferation and cytokine production is completely abrogated after surgical denervation of the spleen. Thus, during conditioning, the CNS learns to reinstate at demand a CsA-like immunosuppression via splenic innervation. This might be used as a supportive therapy for controlling immune functions.

Dialysis patients, waiting for kidney transplantation, were asked about their quality of life. Data from 1027 persons have been collected. Compared to a population sample by the "Nottingham Health Profile" (NHP), dialysis patients showed double the frequency of symptoms--only for the subscale "pain" no significant difference could be recognised. Duration of dialysis treatment reduces the quality of life considerably: increasing troubles have been observed through different quality of life scales. Age shows less important influence concerning "pain" and "physical mobility", even a decrease of symptoms in elder patients has been demonstrated by NHP-subscales for "emotional reaction" and "social isolation". Gender, education, kind of disease and dialysis treatment, and the fact of former transplantations had only marginal influence on some different dimensions of life quality. The study demonstrates in which way the patients perception of life quality could be operational and integrated in analysis and evaluation of therapeutic procedures.