Auswirkungen klinischer und subklinischer Krankheit auf ethologische und klinisch-chemische Merkmale beim Schwein

Ein wichtiger Aspekt für ungestörtes Wohlbefinden und gute Lebensqualität bei Nutztieren ist die Abwesenheit von Krankheit. Während einer Infektion kommt es zu koordinierten Änderungen sowohl des Verhaltens als auch physiologischer Indikatoren, die auf die Präsenz von Krankheit und Leiden hinweisen. Derartige Veränderungen werden im Angelsächsischen mit dem Begriff „sickness behaviour“ beschrieben. Die Folgen sind u.a. eine erhöhte Körpertemperatur und die Entstehung krankheitsspezifischen Verhaltens. Obwohl Änderungen des Verhaltens und physiologischer Vorgänge im Organismus im Falle einer Erkrankung einem zentral motivierten und nützlichen Geschehen entsprechen, dessen Ziel es ist, eine Rekonvaleszenz herbeizuführen, kommt es zu Einschränkungen körperlicher Funktionen und der Integrität des Verhaltens. Dies führt zu einem eingeschränkten Wohlbefinden, aus dem ein Leidenszustand abgeleitet werden kann. Infektionskrankheiten sind im Bereich der Tierproduktion weit verbreitet. Sie stellen ein großes Problem dar. Bisher wurden allerdings nur wenige Studien veröffentlicht, die Auswirkungen spezifischer Krankheiten auf Verhaltensindikatoren beim Schwein detailliert beschreiben: Untersuchungen zu Verhaltensänderungen bei Morbus Aujeszky, Colidiarrhoe nach dem Absetzen und PRRS (Porzines Reproduktives und Respiratorisches Syndrom). In der vorliegenden Studie sollte geprüft werden, inwieweit anhand veränderter Verhaltensindikatoren und deren Bezug zu klinischen, hämatologischen und klinischchemischen Parametern in definierten Krankheitsmodellen die Einschätzung des tierischen Leidens im Zuge von Infektionskrankheiten verbessert werden kann. Basierend auf einem definierten Parasitose-Modell (Sarcocystis miescheriana) wurden die tierartspezifischen Verhaltensparameter Liegen, Aktivität im Liegen, Laufen, Futter- und Wasseraufnahme, Wühlen/Erkunden und Sozialkontakte bei Schweinen der F2-Generation einer Kreuzung aus Meishan und Pietrain während der klassischen Phasen der Infektion beurteilt. Vorteil des ausgewählten Krankheits-Modells ist, dass sich bereits bei einer milden Infektion die unterschiedlichen Krankheitsstadien (akut, subklinisch, chronisch) differenziert darstellen lassen. Anhand von Videoaufzeichnungen wurden die genannten Verhaltensweisen quantifiziert und ausgewertet und mit parallel erhobenen klinischen und labordiagnostischen Parametern in Zusammenhang gebracht. Verglichen mit dem physiologischen Stadium, in dem eine Gesamtaktivität der untersuchten Schweine von 44 % des Beobachtungszeitraumes nachgewiesen wurde, konnte im Akutstadium (Tag 14 p. i.), welches durch den Ablauf der zweiten Schizogonie der Sarcocystose verursacht wird, eine signifikante Änderung des gesamten Verhaltensrepertoires beobachtet werden. Die aktiven Verhaltensindikatoren waren in ihrer zeitlichen Ausprägung auf 10 % der Beobachtungszeit reduziert. Die Phasen des inaktiven Liegens waren dementsprechend verlängert. Im chronischen Stadium der Infektion (Tag 42p.i.) kommt es zur Bildung von Zysten in Skelett- und Herzmuskulatur. In diesem Zeitraum kam es im Vergleich zu den gesunden Schweinen ebenfalls zu einer herabgesetzten Gesamtaktivität von 20 %. Das dazwischen liegende subklinische Krankheitsstadium (Tag 28 p.i.) wies trotz Erholung der Tiere von der akuten Krankheit immernoch signifikante Änderungen des Verhaltensmusters auf (Gesamtaktivität von 33 %). Die Ergebnisse der Verhaltensbeobachtungen deckten sich mit Änderungen der klinischen, hämatologischen und klinisch-chemischen Parameter im akuten, subklinischen und chronischen Stadium der Sarcocystose. Unabhängig vom Infektionsstadium hatten Schweine mit pathologischen klinischen und labordiagnostischen Werten ein 2 bis 5-fach erhöhtes Risiko, ein Verhalten zu entwickeln, das außerhalb des zuvor definierten „Standardbereiches“ der gesunden Population lag. Bemerkenswert waren Verknüpfungen des Verhaltens mit der Körperinnentemperatur sowie u.a. mit Laborparametern wie der Kreatinkinase, der Aspartataminotransferase und den Leukozyten. Vermutet werden könnte auch ein Zusammenhang mit der Aktivität der Alkalischen Phosphatase. Dies wäre aber in weiteren Untersuchungen zu beweisen. Aufgrund der Abweichungen des Verhaltens von den zuvor definierten Standardbedingungen war davon auszugehen, dass die erkrankten Schweine nicht mehr ihrer artspezifischen Bedürfnisbefriedigung nachgehen konnten. Dadurch wurde das Wohlbefinden der Tiere gestört und im Sinne verschiedener Autoren (u.a. Brummer 1978; Sambraus 1981, 1991; Martin 1996; Würbel 2009) ein Leidenszustand induziert. Über die enge Korrelation zu klinischen und labordiagnostischen Parametern könnte in Zukunft eine verbesserte Basis geschaffen werden, um Wohlbefinden und Leiden bei Tieren frühzeitig und objektiver zu erfassen und zu quantifizieren.The absence of disease is important for the welfare and quality of life of farm animals. During an infection in such animals there are both behavioural and physiological indicators of sickness and suffering. These coordinated changes are referred to as “sickness behaviour”. The results include an increased body temperature and the existence of sickness related behaviour. Changes in the behaviour and physiological process of the animal in the event of sickness correspond to a centrally motivated and useful state, the goal of which is to allow the organism to reconvalesce. However, the restrictions of bodily function and the integrity of its behaviour can lead to a state of suffering. Infectious diseases are widespread in the area of animal production, and present a large problem. At this stage, very few studies have been published which describe the effects of specific diseases on the behavioural indices of pigs: examinations of changing conditions in Morbus Aujeszky, postweaning colibacillosis and PRRS (Porcine Reproductive and Respiratory Syndrome). This study proposes to examine how changes in the animal´s behaviour and its relationship to clinical and clinical-chemical parameters in defined disease models can improve the assessment of the animal´s suffering from infectious disease. Based on a well-defined model disease (Sarcocystis miescheriana) we have studied the behavioural patterns of F2 Meishan x Pietrain crossbred pigs in various stages of health, acute disease, recovery and chronic disease. Prime advantage of the model is the ability to induce a definite, but relatively mild clinical infection with expression of the three distinct stages of Sarcocystosis. Data showing the pigs lying inactive, activity during lying, feeding and drinking, walking, rooting and social interactions were captured from video records and associated with clinical/clinical-chemical parameters. In comparison to the physiological stage, where the observed pigs were active 44% of the studied period, in the acute stage (day 14; during second-generation schizogony) a significant change in the overall behaviour was observed: total activity of the pigs was reduced to 10% of the observed time. The phases of inactive lying were accordingly extended. In the chronic stage of infection (day 42) encystations develop in the skeletal muscles and the heart. During this stage, compared to healthy pigs, the total activity was reduced to 20%. Between these two stages, the pigs start to recover from acute disease and become subclinical (day 35). Despite this recovery, the pigs still show significant changes in behaviour (total activity of 33%). The results of the observed behaviour were in agreement with most patterns of clinical and clinical-chemical parameters in the acute, subclinical and chronical stages of Sarcocystosis. Independent of the stages of infection, pigs with pathological clinical and clinical-chemica

Simplification of biochemical models: a general approach based on the analysis of the impact of individual species and reactions on the systems dynamics

Background: Given the complex mechanisms underlying biochemical processes systems biology researchers tend to build ever increasing computational models. However, dealing with complex systems entails a variety of problems, e.g. difficult intuitive understanding, variety of time scales or non-identifiable parameters. Therefore, methods are needed that, at least semi-automatically, help to elucidate how the complexity of a model can be reduced such that important behavior is maintained and the predictive capacity of the model is increased. The results should be easily accessible and interpretable. In the best case such methods may also provide insight into fundamental biochemical mechanisms. Results: We have developed a strategy based on the Computational Singular Perturbation (CSP) method which can be used to perform a "biochemically-driven" model reduction of even large and complex kinetic ODE systems. We provide an implementation of the original CSP algorithm in COPASI (a COmplex PAthway SImulator) and applied the strategy to two example models of different degree of complexity - a simple one-enzyme system and a full-scale model of yeast glycolysis. Conclusion: The results show the usefulness of the method for model simplification purposes as well as for analyzing fundamental biochemical mechanisms. COPASI is freely available at http://www.copasi.org

Minimum inhibitory (MIC) and minimum microbicidal concentration (MMC) of polihexanide and triclosan against antibiotic sensitive and resistant Staphylococcus aureus and Escherichia coli strains

Background: An in-vitro study was conducted investigating the antimicrobial efficacy of polihexanide and triclosan against clinical isolates and reference laboratory strains of Staphylococcus aureus and Escherichia coli

Erythropoietin and the effect of oxygen during proliferation and differentiation of human neural progenitor cells

<p>Abstract</p> <p>Background</p> <p>Hypoxia plays a critical role in various cellular mechanisms, including proliferation and differentiation of neural stem and progenitor cells. In the present study, we explored the impact of lowered oxygen on the differentiation potential of human neural progenitor cells, and the role of erythropoietin in the differentiation process.</p> <p>Results</p> <p>In this study we demonstrate that differentiation of human fetal neural progenitor cells under hypoxic conditions results in an increased neurogenesis. In addition, expansion and proliferation under lowered oxygen conditions also increased neuronal differentiation, although proliferation rates were not altered compared to normoxic conditions. Erythropoietin partially mimicked these hypoxic effects, as shown by an increase of the metabolic activity during differentiation and protection of differentiated cells from apoptosis.</p> <p>Conclusion</p> <p>These results provide evidence that hypoxia promotes the differentiation of human fetal neural progenitor cells, and identifies the involvement of erythropoietin during differentiation as well as different cellular mechanisms underlying the induction of differentiation mediated by lowered oxygen levels.</p

Impact of carbohydrate-reduced nutrition in septic patients on ICU: study protocol for a prospective randomised controlled trial

Introduction: Sepsis is defined as detrimental immune response to an infection. This overwhelming reaction often abolishes a normal reconstitution of the immune cell homeostasis that in turn increases the risk for further complications. Recent studies revealed a favourable impact of ketone bodies on resolution of inflammation. Thus, a ketogenic diet may provide an easy-to-apply and cost-effective treatment option potentially alleviating sepsis-evoked harm. This study is designed to assess the feasibility, efficiency and safety of a ketogenic diet in septic patients. Methods and analysis: This monocentric study is a randomised, controlled and open-label trial, which is conducted on an intensive care unit of a German university hospital. As intervention enteral nutrition with reduced amount of carbohydrates (ketogenic) or standard enteral nutrition (control) is applied. The primary endpoint is the detection of ketone bodies in patients' blood and urine samples. As secondary endpoints, the impact on important safety-relevant issues (eg, glucose metabolism, lactate serum concentration, incidence of metabolic acidosis, thyroid function and 30-day mortality) and the effect on the immune system are analysed. Ethics and dissemination The study has received the following approvals: Ethics Committee of the Medical Faculty of Ruhr-University Bochum (No. 18-6557-BR). Results will be made available to critical care survivors, their caregivers, the funders, the critical care societies and other researchers by publication in a peer-reviewed journal

Neutrophil Granulocytes in Ovarian Cancer - Induction of Epithelial-To- Mesenchymal-Transition and Tumor Cell Migration

Background: Ovarian cancer (OvCa) is a highly aggressive malignoma with a tumor-promoting microenvironment. Infiltration of polymorphonuclear neutrophils (PMN) is frequently seen, raising the question of their impact on tumor development. In that context, effects of PMN on human ovarian cancer cells were assessed. Methods: Human epithelial ovarian cancer cells were incubated with human PMN, lysate of PMN, or neutrophil elastase. Morphological alterations were observed by time-lapse video-microscopy, and the underlying molecular mechanism was analyzed by flow cytometry and Western blotting. Functional alternations were assessed by an in vitro wound healing assay. In parallel, a large cohort of n=334 primary OvCa tissue samples of various histological subtypes was histologically evaluated. Results: Co-cultivation of cancer cells with either PMN or PMN lysate causes a change of the polygonal epithelial phenotype of the cells towards a spindle shaped morphology, causing a cribriform cell growth. The PMN-induced alteration could be attributed to elastase, a major protease of PMN. Elastase-induced shape change was most likely due to the degradation of membranous E-cadherin, which results in loss of cell contacts and polarity. Moreover, in response to elastase, epithelial cytokeratins were downmodulated, in parallel with a nuclear translocation of β-catenin. These PMN-elastase induced alterations of cells are compatible with an epithelial-to-mesenchymal transition (EMT) of the cancer cells. Following EMT, the cells displayed a more migratory phenotype. In human biopsies, neutrophil infiltration was seen in 72% of the cases. PMN infiltrates were detected preferentially in areas with low E-cadherin expression. Conclusion: PMN in the microenvironment of OvCa can alter tumor cells towards a mesenchymal and migratory phenotype

HepatoNet1: a comprehensive metabolic reconstruction of the human hepatocyte for the analysis of liver physiology

We present HepatoNet1, a manually curated large-scale metabolic network of the human hepatocyte that encompasses >2500 reactions in six intracellular and two extracellular compartments.Using constraint-based modeling techniques, the network has been validated to replicate numerous metabolic functions of hepatocytes corresponding to a reference set of diverse physiological liver functions.Taking the detoxification of ammonia and the formation of bile acids as examples, we show how these liver-specific metabolic objectives can be achieved by the variable interplay of various metabolic pathways under varying conditions of nutrients and oxygen availability

Exploring the gap: attitudes, knowledge, and training needs in complementary and integrative medicine among healthcare professionals at German university hospitals

IntroductionThe use of Complementary and Integrative Medicine (CIM) is very popular among the general population in Germany. However, international studies show that nurses, physicians, and other health care professionals (HCPs) at hospitals often do not feel sufficiently informed about different CIM approaches. Moreover, they do not feel trained enough to counsel their patients appropriately. In the German-speaking context, particularly within university hospitals, research on this subject is scarce. Therefore, the aim of this explorative study was to evaluate attitudes, subjective knowledge, and needs regarding CIM among HCPs with direct patient interaction across all four university hospitals in the federal state of Baden-Württemberg, Germany (Tübingen, Ulm, Freiburg, Heidelberg).MethodsThe multicenter, cross-sectional, anonymous full survey was conducted online using a self-developed, semi-structured, web-based questionnaire. Recruitment took place via all-inclusive e-mail distribution lists of all four university hospitals.ResultsA total of n = 2,026 participants (response rate varied by location from about 5 to 14%) fully answered the questionnaire. Nurses constituted the largest professional group (n = 1,196; 59%), followed by physicians (n = 567; 28%), physiotherapists (n = 54), psychologists (n = 48), midwives (n = 37), and other professions (n = 124). More than two-thirds (71%, n = 1,437) of the participants were female and 14% (n = 286) reported additional training in CIM. The overall attitude toward CIM (10-point Likert scale, 10 = “very favorable”) was clearly positive (M ± SD: 7.43 ± 2.33), with notable differences between professional groups: midwives (9.05 ± 1.18), physiotherapists (8.44 ± 1.74), and nurses (8.08 ± 1.95) expressed the highest support, whereas physicians (5.80 ± 2.39) the lowest. 42% of the participants incorporated CIM in patient care (from 33% of physicians to 86% of midwives). Overall, relaxation therapy (n = 1,951; 96%), external applications (n = 1,911; 94%), massage (n = 1,836; 91%), and meditation/mindfulness (n = 1,812; 89%) were rated as useful or rather useful for patients. The average self-assessed knowledge level about CIM was moderate (M ± SD: 5.83 ± 2.03). Most of the participants found CIM training at university hospitals important and saw research about CIM as one of the tasks of university hospitals. The participants expressed the highest interest in education for acupuncture/acupressure, relaxation therapies, and manual medicine.DiscussionThis comprehensive survey of health care professionals (HCPs) at university hospitals in Germany reveals a clearly positive disposition toward CIM, aligning with findings from other hospital-based surveys and highlighting differences among professional groups. While most therapies deemed beneficial for patient care are supported by positive evidence, further research is required for others. Given the average self-reported knowledge of CIM, targeted education is essential to meet the needs of both HCPs and patients and to ensure the provision of evidence-based information on the risks and benefits of CIM

Sulforaphane Inhibits Inflammatory Responses of Primary Human T-Cells by Increasing ROS and Depleting Glutathione

The activity and function of T-cells are influenced by the intra- and extracellular redox milieu. Oxidative stress induces hypo responsiveness of untransformed T-cells. Vice versa increased glutathione (GSH) levels or decreased levels of reactive oxygen species (ROS) prime T-cell metabolism for inflammation, e.g., in rheumatoid arthritis. Therefore, balancing the T-cell redox milieu may represent a promising new option for therapeutic immune modulation. Here we show that sulforaphane (SFN), a compound derived from plants of the Brassicaceae family, e.g., broccoli, induces a pro-oxidative state in untransformed human T-cells of healthy donors or RA patients. This manifested as an increase of intracellular ROS and a marked decrease of GSH. Consistently, increased global cysteine sulfenylation was detected. Importantly, a major target for SFN-mediated protein oxidation was STAT3, a transcription factor involved in the regulation of TH17-related genes. Accordingly, SFN significantly inhibited the activation of untransformed human T-cells derived from healthy donors or RA patients, and downregulated the expression of the transcription factor RORγt, and the TH17-related cytokines IL-17A, IL-17F, and IL-22, which play a major role within the pathophysiology of many chronic inflammatory/autoimmune diseases. The inhibitory effects of SFN could be abolished by exogenously supplied GSH and by the GSH replenishing antioxidant N-acetylcysteine (NAC). Together, our study provides mechanistic insights into the mode of action of the natural substance SFN. It specifically exerts TH17 prone immunosuppressive effects on untransformed human T-cells by decreasing GSH and accumulation of ROS. Thus, SFN may offer novel clinical options for the treatment of TH17 related chronic inflammatory/autoimmune diseases such as rheumatoid arthritis

Glucosylsphingosine Is a Highly Sensitive and Specific Biomarker for Primary Diagnostic and Follow-Up Monitoring in Gaucher Disease in a Non-Jewish, Caucasian Cohort of Gaucher Disease Patients

Gaucher disease (GD) is the most common lysosomal storage disorder (LSD). Based on a deficient β-glucocerebrosidase it leads to an accumulation of glucosylceramide. Standard diagnostic procedures include measurement of enzyme activity, genetic testing as well as analysis of chitotriosidase and CCL18/PARC as biomarkers. Even though chitotriosidase is the most well-established biomarker in GD, it is not specific for GD. Furthermore, it may be false negative in a significant percentage of GD patients due to mutation. Additionally, chitotriosidase reflects the changes in the course of the disease belatedly. This further enhances the need for a reliable biomarker, especially for the monitoring of the disease and the impact of potential treatments.Here, we evaluated the sensitivity and specificity of the previously reported biomarker Glucosylsphingosine with regard to different control groups (healthy control vs. GD carriers vs. other LSDs).Only GD patients displayed elevated levels of Glucosylsphingosine higher than 12 ng/ml whereas the comparison controls groups revealed concentrations below the pathological cut-off, verifying the specificity of Glucosylsphingosine as a biomarker for GD. In addition, we evaluated the biomarker before and during enzyme replacement therapy (ERT) in 19 patients, demonstrating a decrease in Glucosylsphingosine over time with the most pronounced reduction within the first 6 months of ERT. Furthermore, our data reveals a correlation between the medical consequence of specific mutations and Glucosylsphingosine.In summary, Glucosylsphingosine is a very promising, reliable and specific biomarker for GD