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Bacterial infections are one of the most important clinical problems and contribute to the high mortality rate in patients with decompensated liver cirrhosis. Fluoroquinolones are highly effective drugs and are frequently used in patients with cirrhosis of the liver, especially for prophylaxis of spontaneous[for full text, please go to the a.m. URL], Infektiologie Update 2016; 25. Jahrestagung der Paul-Ehrlich-Gesellschaft für Chemotherapie (PEG)

Einführung: Die Polyneuropathie bei Dialysepatienten ist multifaktoriell – insbesondere wird sie durch Urämietoxine, Bluthochdruck, Ischämien und Diabetes begünstigt. Ziel dieser Arbeit ist es, Änderungen der Hornhautnerven bei Dialyse-Patienten in Abhängigkeit vom[for full text, please go to the a.m. URL], 65. Tagung der Vereinigung Norddeutscher Augenärzte

Background/Case Studies: Donor granulocyte concentrates (GCs) are limited for broad application, mainly due to a short storage time of 6 ato 24 hours after donation, and they have to be prepared specifically for each patient, in compliance with blood group and cytomegalovirus (CMV) status. This storage period partially results from the high contamination (approx. 99%) with RBCs and PLTs. We developed a closed-system method compatible with standard blood bank technologies to enrich the granulocytes and store them for longer periods. Study Design/Methods: GCs were prepared according to clinical standard procedures by apheresis in a medical blood bank. About 18 hours before apheresis, the donors were stimulated with steroids and G-CSF. These standard GCs were sedimented on hydroxyethylstarch, washed twice with 0.9% NaCl-solution, and resuspended in ABOidentical filtered plasma. Procedure was done in a closed multi-bag disposable system. These purified GCs (pGCs) were stored in platelet storage bags at a cell concentration of 5 x 107 leukocytes/mL without agitation at room temperature for up to 72 hours in 200 or 400 mL. Cell count and viability of the WBCs were tested daily. Phagocytosis and oxyburst were analyzed by flow cytometry. Results/Findings: The patented method resulted in a significant reduction of RBCs (98%, by sedimentation) and PLTs (95%, by washing) and in a highly purified WBC population and enrichment of granulocytes. In addition, the viability was significantly improved. Both the percentage of phagocytic and oxidizing PMNs as well as the phagocytic capacity remained stable (approx. 100%). After 72 hours of storage, 94% of the initial number of phagocytes remained. Furthermore, the viability of these cells improved significantly (0 hr: 86%; 72 hr: 92%), most probably due to phagocytosis of dead cells. Both, the percentage of phagocytic granulocytes and the phagocytosis intensity (mean fluorescence intensity due to ingestion of FITC-labeled E. coli) remained stable over the period of storage. After 72 hours, 96% of the granulocytes were functional and showed unimpaired phagocytic intensity. Similar effects were observed in the oxyburst tests. Almost 100% of the stored granulocytes were fully functional. Based on the results measured by flow cytometry and chemiluminscence, after 72 hours the oxyburst output of the stored granulocytes was comparable to the oxyburst level of the nonpurified GCs. Conclusion: By the improvement of the purification and storage methods and the resulting high purity of the granulocyte concentrates, an extension of application time from 24 to 72 hours can be achieved. Furthermore, all procedures of purification and storage of granulocyte concentrates can be done in a closed system compliant with GMP regulatory requirements. These two important facts may broaden the applicability of granulocyte transfusions in medicine.

Objective: Invasive fungal infections are associated with a high mortality rate. Systemic antimycotics for the treatment are poorly investigated for drug-induced hepatotoxicity. Therefore, we used an established in-vitro test with a standardised permanent cell line and tested the hepatotoxicity of acetaminophen and of systemic antimycotics. Methods: In a standardised mikrotiterplate assay the toxicity of different concentrations of acetaminophen (AM 7.62 mM | 15.24 mM) and of the antimycotics Anidulafungin (Ani), Liposomal Amphotericerin B (L-AmB), Fluconazole (Fluco)) were tested with human liver cells (HepG2/C3A, 500,000 cells/well; medium). The lowest test concentration of antimycotics was the mean plasma levels after induction of an i.v. therapy (Cmax), as well as the 5-times and 10-times concentrations of Cmax were analyzed. As controls served plasma with the different agents and a plasma plus medium control without agents (Control Group = CG). After incubation time of 2 × 3 days the standard parameters of vitality (e.g. LDH release, trypan blue staining) and DNA fragmentation evaluation by flow cytometry-based terminal transferase dUTP nick end-labeling (TUNEL) assay were tested. Results: (Median, 25/75 percentile): All antimycotics and AM showed a decreased vitality [%] (L-AmB 73, 66/77 | Ani 80, 76/82 | Fluco 75, 72/79) in the Cmax-concentration compared with the CG (94, 92/95) with corresponding increased LDH release [U/l] (L-AmB 70, 63.5/90.25 | Ani 66, 58.5/82.5 | Fluco 144, 107.75/182.25 | CG 70.5, 54/97 | AM 192, 163.75/214.5). With increasing concentrations of Ani a markedly decrease of cell vitality was seen; in contrast, Fluco und L-AmB showed a slightly increase. Comparing to the CG, AM, Fluco and L-AmB led to a significant lower number of DNA positive cells [%] (L-AmB 0.6, 0.2/1.1 | Fluco 4.9, 4.5/5.3 | CG 3.7, 1.7/8.4 | AM 2.3, 1.4/17.4). The number of DNA positive cells increased significantly in the Ani-group (10 × Cmax 58, 48.1/70). Conclusion: All antimycotics showed a slightly hepatotoxic potential in the Cmax concentration; Ani a stronger hepatotoxicity in higher concentrations. Therapeutic drug monitoring would be helpful for the use in critically ill patients.

INTRODUCTION. Nearly 19 % of patients with severe sepsis developed liver dysfunction or failure in the course of disease (1). The late diagnosis of liver failure is a major problem in critically ill patients. Early diagnosis of liver dysfunction and failure can enable early onset of therapy and may lead to an improvement of prognosis of these patients. OBJECTIVES. To verify the clinical relevance of a new test device for early diagnosis of liver failure, we conducted a second study with severe septic patients. A first study revealed, that plasma from septic patients lead to an impairment of viability and function of sensorcells (2). METHODS. We developed a new test device for early diagnosis of liver failure (patent pending). The basic test compounds consist of human liver cells (HepG2/C3A). In a standardised mikrotiterplate assay the toxicity of patient plasma was tested (500.000 cells/ well). After incubation with plasma from patients the viability of cells (XTT test, Trypan blue-staining), the cytochrome 1A2 activity (metabolism of etoxyresorufin) and synthesis of albumin are measured. In a prospective clinical study in 99 ICU-patients two test groups were studied: the septic shock group (n = 51, SSG) and the non-septic shock group (n = 49, NSG) as control group. At time of inclusion, after 3 and 7 days, 10 ml blood was drawn from the patients for testing with the cytotoxicity test. Patients were followed up for hospital survival. The results are expressed as median and range. Differences were considered significant at p\0.05. Significance was determined with the Mann-Whitney U-test. RESULTS. The in-hospital mortality was 23.5 % in the SSG; one patient of the NSG died. The APACHE II-scores at ICU arrival were 32 (15) in the SSG and 9 (10) in the NSG, the SOFA-scores at inclusion were 13 (6.8) in the SSG and 2 (4.6) in the NSG. The values of bilirubin (SSG-Survivors: 19.2 lmol/l (26.6), SSG-Non-Survivors: 21.4 (88.2), NSG: 15.2 (17)) were significantly higher and the Prothrombin-time lower in the SSG (Survivors: 73 % (32.8), Non-Survivors: 66 (22.8)) than in the NSG 90 (25.8). The plasma of patients with septic shock impaired significantly viability and cellular functions of HepG2/C3A cells in all parameters compared with the plasma of non-septic patients. These effects were more pronounced with plasma of non-survivors in the SSG at inclusion, after 3 and 7 days (data not shown). CONCLUSIONS. The presented liver cells based biosensor showed hepatotoxicity of plasma from patients with septic shock in a second study; moreover, these effects were more pronounced in non-survivors. The new test may contribute to an early diagnosis of liver failure in septic patients.