In Vivo and In Vitro Characterization of Primary Human Liver Macrophages and Their Inflammatory State

Liver macrophages (LMs) play a central role in acute and chronic liver pathologies. Investigation of these processes in humans as well as the development of diagnostic tools and new therapeutic strategies require in vitro models that closely resemble the in vivo situation. In our study, we sought to gain further insight into the role of LMs in different liver pathologies and into their characteristics after isolation from liver tissue. For this purpose, LMs were characterized in human liver tissue sections using immunohistochemistry and bioinformatic image analysis. Isolated cells were characterized in suspension using FACS analyses and in culture using immunofluorescence staining and laser scanning microscopy as well as functional assays. The majority of our investigated liver tissues were characterized by anti-inflammatory LMs which showed a homogeneous distribution and increased cell numbers in correlation with chronic liver injuries. In contrast, pro-inflammatory LMs appeared as temporary and locally restricted reactions. Detailed characterization of isolated macrophages revealed a complex disease dependent pattern of LMs consisting of pro- and anti-inflammatory macrophages of different origins, regulatory macrophages and monocytes. Our study showed that in most cases the macrophage pattern can be transferred in adherent cultures. The observed exceptions were restricted to LMs with pro-inflammatory characteristics

The Value of Graft Implantation Sequence in Simultaneous Pancreas-Kidney Transplantation on the Outcome and Graft Survival

Background/Objectives: The sequence of graft implantation in simultaneous pancreas-kidney transplantation (SPKT) warrants additional study and more targeted focus, since little is known about the short- and long-term effects on the outcome and graft survival after transplantation. Material and methods: 103 patients receiving SPKT in our department between 1999 and 2015 were included in the study. Patients were divided according to the sequence of graft implantation into pancreas-first (PF, n = 61) and kidney-first (KF, n = 42) groups. Clinicopathological characteristics, outcome and survival were reviewed retrospectively. Results: Donor and recipient characteristics were similar. Rates of post-operative complications and graft dysfunction were significantly higher in the PF group compared with the KF group (episodes of acute rejection within the first year after SPKT: 11 (18%) versus 2 (4.8%); graft pancreatitis: 18 (18%) versus 2 (4.8%), p = 0.04; vascular thrombosis of the pancreas: 9 (14.8%) versus 1 (2.4%), p = 0.03; and delayed graft function of the kidney: 12 (19.6%) versus 2 (4.8%), p = 0.019). The three-month pancreas graft survival was significantly higher in the KF group (PF: 77% versus KF: 92.1%; p = 0.037). No significant difference was observed in pancreas graft survival five years after transplantation (PF: 71.6% versus KF: 84.8%; p = 0.104). Kidney graft survival was similar between the two groups. Multivariate analysis revealed order of graft implantation as an independent prognostic factor for graft survival three months after SPKT (HR 2.6, 1.3–17.1, p = 0.026) and five years (HR 3.7, 2.1–23.4, p = 0.040). Conclusion: Our data indicates that implantation of the pancreas prior to the kidney during SPKT has an influence especially on the early-post-operative outcome and survival rate of pancreas grafts

Placebo from an enactive perspective

Due to their complexity and variability, placebo effects remain controversial. We suggest this is also due to a set of problematic assumptions (dualism, reductionism, individualism, passivity). We critically assess current explanations and empirical evidence and propose an alternative theoretical framework—the enactive approach to life and mind—based on recent developments in embodied cognitive science. We review core enactive concepts such as autonomy, agency, and sense-making. Following these ideas, we propose a move from binary distinctions (e.g., conscious vs. non-conscious) to the more workable categories of reflective and pre-reflective activity. We introduce an ontology of individuation, following the work of Gilbert Simondon, that allow us to see placebo interventions not as originating causal chains, but as modulators and triggers in the regulation of tensions between ongoing embodied and interpersonal processes. We describe these interrelated processes involving looping effects through three intertwined dimensions of embodiment: organic, sensorimotor, and intersubjective. Finally, we defend the need to investigate therapeutic interactions in terms of participatory sense-making, going beyond the identification of individual social traits (e.g., empathy, trust) that contribute to placebo effects. We discuss resonances and differences between the enactive proposal, popular explanations such as expectations and conditioning, and other approaches based on meaning responses and phenomenological/ecological ideas

Die nicht-invasive Messung der Steatohepatitis korreliert mit einem erhöhten Risiko für koronare Herzerkrankungen

Die Fettlebererkrankung (fatty liver disease; FLD) und koronare Herzkrankheit (KHK) stellen zwei wichtige Manifestationen des metabolischen Syndroms (MS) dar. Eine frühzeitige Erkennung sowie Prävention dieser beiden Entitäten ist eine große Herausforderung in der Gesundheitsversorgung der heutigen Zeit. So empfehlen die aktuellen Leitlinien zur Nichtalkoholischen Fettlebererkrankung (NAFLD) ein Screening dieser Patient:innen auf das Vorliegen einer KHK, sowie Patient:innen mit KHK auf eine FLD zu untersuchen. Nichtsdestotrotz sind derartige Empfehlungen in den KHK-Leitlinien kaum zu finden. Eine Verbindung zwischen FLD und KHK besteht aufgrund gemeinsamer Risikofaktoren (Diabetes mellitus, Übergewicht etc.), jedoch mehren sich die Anzeichen eines kausalen Zusammenhangs, welcher Gegenstand aktueller Forschung ist. So wird unter anderem ein Zusammenhang zwischen entzündlicher Aktivität und Atherosklerose vermutet. Die Arbeitsgruppe der Universitätsklinik Leipzig hat sich in der vorliegenden Studie mit der Frage beschäftig, ob mittels nichtinvasiver Untersuchungsverfahren zur Messung der Fettleber, Leberfibrose und Steatohepatitis bei einer kardiologischen Kohorte eine Aussage zum Risiko einer interventionspflichtigen KHK getroffen werden kann. Insgesamt stellte die FLD mit einer Prävalenz von ca. 40% ein häufiges Krankheitsbild in der untersuchten Kohorte dar. Ein signifikanter Zusammenhang zwischen FLD und KHK ließ sich nicht nachweisen. Auch war der Anteil an Patient:innen mit höhergradiger Fibrose mit ca. 5% vergleichsweise niedrig. Interessanterweise waren die traditionellen Risikofaktoren Diabetes, Übergewicht und arterielle Hypertonie weder mit einer fortgeschrittenen Lebererkrankung noch mit der Notwendigkeit einer kardiovaskulären Intervention signifikant assoziiert, was möglicherweise auf Grund der Fallzahl und des vorselektionierten Patientengutes begründet werden kann. Nur das männliche Geschlecht stand in unserer Kohorte in signifikantem Zusammenhang mit dem KHK-Risiko, was mit den berichteten Daten übereinstimmt und auf die Praxis der Auswahl der Patient:innen zum Zeitpunkt der Rekrutierung zurückzuführen ist, bei der Frauen eine höhere Rate an falsch-positiven Ergometrie- und Myokardperfusionsszintigraphien aufweisen. Auch die gängigen Labor-Tests - in der vorliegenden Studie der NAFLD-Fibrosis-Score (NFS) und FIB4-Index - zeigten keine signifikante Assoziation zum Ergebnis der HKU. In unserer Studie konnte ein signifikanter Zusammenhang zwischen erhöhter Leber-Steifigkeit gemessen mittels Scherwellen-Elastografie (vibration-controlled transient elastography; VCTE) und interventionspflichtiger KHK nachgewiesen werden (wenn auch unterhalb der gängigen Cut-offs einer signifikanten Fibrose). Diese erhöhten Steifigkeitswerte unterhalb der pathologischen Grenze reflektieren nicht zwingend eine Gewebsfibrose, können aber Ausdruck einer Entzündung und damit verbundenen veränderten Vaskularisation sein. Dies spiegelt sich in der Berechnung des FAST-Scores wider. Der FAST-score stellt einen Surrogat-Marker für die Steatohepatitis dar, der das Risiko einer fibrotischen Steatohepatitis mit guter Genauigkeit abbildet und sich aus Werten der VCTE sowie der Aspartat-Aminotransferase (AST) berechnet. In unserer Kohorte war der FAST-Score der stärkste Prädiktor für eine interventionsbedürftige KHK. Selbst nach Berücksichtigung anderer Risikofaktoren in der multivariaten Analyse stellte sich der FAST-Score als unabhängiger Risikofaktor einer KHK heraus. Vergleichbare Beobachtungen wurden unseres Wissenstands nach bisher nicht publiziert. Daher müssen diese Ergebnisse in weiteren multizentrischen Studien bestätigt werden. Die Inflammation könnte also das Bindeglied zwischen Steatohepatitis und KHK sein, was im Einklang mit den möglichen Pathomechanismen, insbesondere dem oxidativen Stress, stehen würde. Es werden somit weitere Studien benötigt, welche sich auf die potenzielle Rolle des FAST-Scores als Stratifizierungsmethode nicht nur im Bereich der Lebererkrankungen, sondern auch bei der Beurteilung kardiovaskulärer Risiko-Patient:innen konzentrieren. Idealerweise könnte dies mit einer Analyse des Lebergewebes, des viszeralen Fettgewebes und der Endothelfunktion einhergehen, um weitere Erkenntnisse über die wechselseitigen pathophysiologischen Mechanismen dieser Erkrankungen zu gewinnen. In der Zusammenschau zeigen unsere Daten, dass die Prävalenz einer fortgeschrittenen FLD bei Patient:innen mit vermuteter KHK gering ist. Nichtinvasive Tests des Fibrose- und Steatosegrades waren nicht mit der Notwendigkeit einer koronaren Intervention verbunden. Erhöhte FAST-Score-Werte unterstreichen die pathophysiologische Bedeutung der Entzündungsaktivität sowohl bei FLD als auch bei KHK.:1 Einführung 1.1 Bibliografische Beschreibung 1.2 Einleitung 1.2.1 Metabolisches Syndrom 1.2.2 Koronare Herzkrankheit 1.2.3 Fettlebererkrankung und Steatohepatitis 1.2.4 Diagnostik von Fettlebererkrankung und Steatohepatitis 1.2.5 Zusammenhang zwischen koronarer Herzkrankheit und Fettlebererkrankung/Steatohepatitis 1.3 Rationale und Ziele der publizierten Studie 2 Publikation 3 Zusammenfassung der Arbeit 4 Literatur 5 Abkürzungen 6 Anlagen 6.1 Lebenslauf 6.2 Wissenschaftliche Veröffentlichungen 6.3 Publikation 6.4 Darstellung des eigenen Beitrags 6.5 Abdruckerlaubnis 6.6 Einreichungserklärung 6.7 Eigenständigkeitserklärung 6.8 Vorbehaltlichkeitserklärung 6.9 Teilnahmebescheinigung: Vorlesung zur “Guten wissenschaftlichen Praxis“ an der Medizinischen Fakultät der Universität Leipzig 6.10 Danksagun

The Effects of Oral Anticoagulant Exposure on the Surgical Outcomes of Patients Undergoing Surgery for High-Risk Abdominal Emergencies

Background!#!In chronic anticoagulant users undergoing surgery, bleeding and thromboembolism are common and serious complications. Many studies on mainly elective or minor emergency surgical procedures with low associated risks have focused on these outcomes. In comparison, patients undergoing high-risk emergency abdominal surgical procedures have not received sufficient attention. This study aimed to compare outcomes between oral anticoagulant users and nonusers who required emergency laparotomy for high-risk abdominal emergencies.!##!Methods!#!Patients who underwent surgery for abdominal emergencies at our institution between January 2012 and July 2019 were retrospectively reviewed.!##!Results!#!There were 875 patients, including 370 anticoagulant users and 505 nonusers. Of the 370 anticoagulant users, 189 (51.3), 77 (20.8%), 45 (12.2%), and 59 (15.9%) were prescribed antiplatelets, a vitamin k antagonist, a direct oral anticoagulant, and a combination drug regimen, respectively. The most common high-risk emergencies requiring surgery in both groups were perforated viscus (25.7% vs 40.9%), mesenteric ischemia with enteric necrosis (27% vs 12.8%), and bowel obstruction (17.6% vs 28.1%). The overall bleeding rate was higher (29.2% vs 22%, p = 0.015) in anticoagulant users than in nonusers, but the major bleeding rate was similar (17.8% vs 14.1%, p = 0.129) between the two groups. The rates of thromboembolic events and mortality were significantly higher in anticoagulant users than in nonusers (25.7% vs 9.7%, p < 0.0001 and 39.7% vs 31.1%, p = 0.01, respectively). Liver cirrhosis, peripheral arterial diseases, reoperation, and blood product transfusion were independent predictors of the overall risk of bleeding or TEEs, according to the multivariate analysis. In this model, liver cirrhosis had the largest overall effect on mortality, followed by pneumonia, thromboembolism, peripheral arterial disease, blood product transfusion, and atrial fibrillation. The use of oral anticoagulants was not an independent predictor of either bleeding or in-hospital mortality. The use of oral anticoagulants was associated with a decreased risk of all-cause in-hospital mortality.!##!Conclusion!#!Based on our results, the continued use of oral anticoagulants is more protective than harmful considering the overall outcomes in this subset of patients

Primary-like Human Hepatocytes Genetically Engineered to Obtain Proliferation Competence as a Capable Application for Energy Metabolism Experiments in In Vitro Oncologic Liver Models

Non-alcoholic fatty liver disease (NAFLD), characterized by lipid accumulation in the liver, is the most common cause of liver diseases in Western countries. NAFLD is a major risk factor for developing hepatocellular carcinoma (HCC); however, in vitro evaluation of hepatic cancerogenesis fails due to a lack of liver models displaying a proliferation of hepatocytes. Originally designed to overcome primary human hepatocyte (PHH) shortages, upcyte hepatocytes were engineered to obtain continuous proliferation and, therefore, could be a suitable tool for HCC research. We generated upcyte hepatocytes, termed HepaFH3 cells, and compared their metabolic characteristics to HepG2 hepatoma cells and PHHs isolated from resected livers. For displaying NAFLD-related HCCs, we induced steatosis in all liver models. Lipid accumulation, lipotoxicity and energy metabolism were characterized using biochemical assays and Western blot analysis. We showed that proliferating HepaFH3 cells resemble HepG2, both showing a higher glucose uptake rate, lactate levels and metabolic rate compared to PHHs. Confluent HepaFH3 cells displayed some similarities to PHHs, including higher levels of the transaminases AST and ALT compared to proliferating HepaFH3 cells. We recommend proliferating HepaFH3 cells as a pre-malignant cellular model for HCC research, while confluent HepaFH3 cells could serve as PHH surrogates for energy metabolism studies

Summary of patient recruitment.

NYHA New York Heart Association, EF Ejection fraction; LSM Liver stiffness measurement.</p

Characterization of patients at high and low risk of relevant liver fibrosis.

Characterization of patients at high and low risk of relevant liver fibrosis.</p

Baseline characteristics of the total cohort stratified for the presence or absence of MAFLD.

Baseline characteristics of the total cohort stratified for the presence or absence of MAFLD.</p