Altered Cytokine Expression and Barrier Properties after In Vitro Infection of Porcine Epithelial Cells with Enterotoxigenic Escherichia coli and Probiotic Enterococcus faecium

The aim of the present study was to elucidate the effects of the probiotic feed additive Enterococcus faecium NCIMB 10415 (E. faecium) on porcine jejunal epithelial cells (IPEC-J2) during an in vitro challenge with enterotoxigenic Escherichia coli (ETEC). Cells were incubated with E. faecium, ETEC, or both, and the effects on barrier function and structure and intra- and intercellular signaling were determined. Coincubation with E. faecium abolished the ETEC- induced decrease in transepithelial resistance (Rt). No differences were seen in the expression levels of the intercellular connecting tight junction proteins examined. However, for the first time, a reorganization of the monolayer was observed in ETEC-infected cells but not in coincubated cells. ETEC induced an increase in cytotoxicity that was prevented by coincubation, whereas apoptosis rates were not affected by bacterial treatment. ETEC increased the mRNA expression and release of proinflammatory cytokines TNF-α, IL-1α, and IL-6 which could be prevented by coincubation for TNF-α mRNA expression and IL-6 protein. Likewise, cAMP concentrations elevated by ETEC were reduced in coincubated cells. These findings indicate a protective effect of the probiotic E. faecium on inflammatory responses during infection with ETEC

The addition of a sagittal image fusion improves the prostate cancer detection in a sensor-based MRI /ultrasound fusion guided targeted biopsy

Background To explore the diagnostic benefit of an additional image fusion of the sagittal plane in addition to the standard axial image fusion, using a sensor-based MRI/US fusion platform. Methods During July 2013 and September 2015, 251 patients with at least one suspicious lesion on mpMRI (rated by PI- RADS) were included into the analysis. All patients underwent MRI/US targeted biopsy (TB) in combination with a 10 core systematic prostate biopsy (SB). All biopsies were performed on a sensor-based fusion system. Group A included 162 men who received TB by an axial MRI/US image fusion. Group B comprised 89 men in whom the TB was performed with an additional sagittal image fusion. Results The median age in group A was 67 years (IQR 61–72) and in group B 68 years (IQR 60–71). The median PSA level in group A was 8.10 ng/ml (IQR 6.05–14) and in group B 8.59 ng/ml (IQR 5.65–12.32). In group A the proportion of patients with a suspicious digital rectal examination (DRE) (14 vs. 29%, p = 0.007) and the proportion of primary biopsies (33 vs 46%, p = 0.046) were significantly lower. The rate of PI-RADS 3 lesions were overrepresented in group A compared to group B (19 vs. 9%; p = 0.044). Classified according to PI-RADS 3, 4 and 5, the detection rates of TB were 42, 48, 75% in group A and 25, 74, 90% in group B. The rate of PCa with a Gleason score ≥7 missed by TB was 33% (18 cases) in group A and 9% (5 cases) in group B; p-value 0.072. An explorative multivariate binary logistic regression analysis revealed that PI-RADS, a suspicious DRE and performing an additional sagittal image fusion were significant predictors for PCa detection in TB. 9 PCa were only detected by TB with sagittal fusion (sTB) and sTB identified 10 additional clinically significant PCa (Gleason ≥7). Conclusion Performing an additional sagittal image fusion besides the standard axial fusion appears to improve the accuracy of the sensor-based MRI/US fusion platform

Standard operating procedures for antibiotic therapy and the occurrence of acute kidney failure

Ein akutes Nierenversagen tritt bei 9 % der hospitalisierten Patienten und bis zu zwei Drit-teln der intensivmedizinisch betreuten Patienten auf. Dadurch werden die Mortalität, die Krankenhausaufenthaltsdauer und gesundheitsökonomischen Kosten erhöht. Das Ziel die-ser Studie war, den Einfluss der Konformität zur den standardisierten Verfahrensanwei-sungen für die Therapie mit Vancomycin, Gentamycin und Tobramycin auf das Auftreten eines akuten Nierenversagens darzustellen. In der klinisch prospektiven Observationsstu-die wurden in zwei separaten Beobachtungszeiträumen 163 erwachsene Patienten erfasst, die länger als 24 Stunden auf einer der drei anästhesiologisch geführten Intensivstationen der Klinik für Anästhesiologie mit Schwerpunkt operative Intensivmedizin der Charité Uni-versitätsmedizin Berlin am Campus Charité Mitte und Campus Virchow-Klinikum aufge-nommen wurden und mit Vancomycin, Gentamicin oder Tobramycin therapiert wurden. Die Gruppeneinteilung in eine Hoch- (HAG) und Niedrigadhärenzgruppe (NAG) erfolgte an-hand eines SOP-Konformitätsrichtwertes von 70 %. Ein akutes Nierenversagen wurde durch die RIFLE-Kriterien klassifiziert. Bezüglich der Basischarakteristika waren die Stu-diengruppen, bis auf einen in der NAG signifikant erhöhten mittleren Aufnahme-APACHE2-Score und eine erniedrigte Häufigkeit von Weichteilinfektionen, homogen. In der LAG (N=75) zeigte sich eine signifikante Erhöhung (p=0,035) der Häufigkeit eines akuten Nie- renversagens mit 36 % gegenüber 21 % in der HAG (N=88). In den univariaten und mul-tivariaten Regressionsanalysen ergab sich die Zugehörigkeit zur NAG als signifikanter Ein-flussfaktor auf die Häufigkeit eines akuten Nierenversagens. Zusammenfassend ist anzunehmen, dass die Qualität der durchgeführten Antibiotikathera-pie mit Vancomycin, Tobramycin und Gentamicin und der Grad der Einhaltung der vorge-gebenen, standardisierten Verfahrensanweisungen wichtige Einflussfaktoren für das Auf-treten eines akuten Nierenversagens darstellen.Objective: Acute kidney failure (AKF) occurs in 9% of hospitalized and in 66% of Intensive Care Unit (ICU) patients. It increases mortality, hospital length of stay, and costs. The aim of this study was to investigate, whether there is an association between adherence to guidelines (standard operating procedures, SOP) for potentially nephrotoxic antibiotics and the occurrence of AKF. Design: Prospective, clinical, non-interventional, observational study Setting: Data collection was performed over a total of 170 days on three ICUs at Charité – Universitaetsmedizin Berlin. Patients: Altogether, 675 patients were included. 163 of them had therapy with vancomycin, gentamicin, or tobramycin; were > 18 years; and treated on ICU for > 24 hours. Pa-tients with an adherence to SOP > 70% were classified into the high adherence group (HAG); patients with an adherence of < than 70 % into the low adherence group (LAG). Definitions: AKF was defined according to RIFLE criteria. Adherence to SOPs was evalu-ated by a retrospective expert audit. Measurements and Main Results: LAG consisted of 75 patients (46%) versus HAG of 88 patients (54%). AKF occurred significantly more in LAG with 36% versus 21% in HAG (p=0,035). Basic characteristics were comparable, except an increased rate of soft tissue infections in LAG. Multivariate analysis revealed an odds ratio of 2, 5- fold for LAG to de-velop AKF compared with HAG (CI 1.038 - 4.437, p=0.039). Conclusion: Low adherence to SOPs was associated with a higher occurrence of AKF

Observational clinical study on the effects of different dosing regimens on vancomycin target levels in critically ill patients: Continuous versus intermittent application

Summary: Different dosing regimens for vancomycin are in clinical use: intermittent infusion and continuous administration. The intention of using these different dosing regimens is to reduce toxicity, to achieve target levels faster and to avoid treatment failure. The aim of this phase IV study was to compare safety and effectiveness in both administration regimens. The study was conducted in 2010 and 2011 in three postoperative intensive care units (ICUs) in a tertiary care university hospital in Berlin, Germany. Adult patients with vancomycin therapy and therapeutic drug monitoring were included. Out of 675 patients screened, 125 received vancomycin therapy, 39% with intermittent and 61% with continuous administration. Patients with continuous administration achieved target serum levels significantly earlier (median day 3 versus 4, p = 0.022) and showed fewer sub-therapeutic serum levels (41% versus 11%, p < 0.001). ICU mortality rate, duration of ICU stay and duration of ventilation did not differ between groups. Acute renal failure during the ICU stay occurred in 35% of patients with intermittent infusion versus 26% of patients with continuous application (p = 0.324). In conclusion, continuous administration of vancomycin allowed more rapid achievement of targeted drug levels with fewer sub-therapeutic vancomycin levels observed. This might indicate that patients with more severe infections or higher variability in renal function could benefit from this form of administration. Keywords: Vancomycin, Intensive care unit, Infection, Therapeutic drug monitorin

Dynamics of urinary calprotectin after renal ischaemia

Background: Urinary calprotectin has been identified as a promising biomarker for acute kidney injury. To date, however, the time-dependent changes of this parameter during acute kidney injury remain elusive. The aim of the present work was to define the time-course of urinary calprotectin secretion after ischaemia/reperfusion-induced kidney injury in comparison to neutrophil gelatinase-associated lipocalin, thereby monitoring the extent of tubular damage in nephron sparing surgery for kidney tumours. Methods: The study population consisted of 42 patients. Thirty-two patients underwent either open or endoscopic nephron sparing surgery for kidney tumours. During the surgery, the renal arterial pedicle was clamped with a median ischaemic time of 13 minutes (interquartile range, 4.5–20.3 minutes) in 26 patients. Ten retro-peritoneoscopic living donor nephrectomy patients and 6 nephron sparing surgery patients in whom the renal artery was not clamped served as controls. Urinary calprotectin and neutrophil gelatinase-associated lipocalin concentrations were repeatedly measured by enzyme-linked immunosorbent assay and assessed according to renal function parameters. Results: Urinary concentrations of calprotectin and neutrophil gelatinase-associated lipocalin increased significantly after ischaemia/reperfusion injury, whereas concentrations remained unchanged after nephron sparing surgery without ischaemia/reperfusion injury and after kidney donation. Calprotectin and neutrophil gelatinase-associated lipocalin levels were significantly increased 2 and 8 hours, respectively, post-ischaemia. Both proteins reached maximal concentrations after 48 hours, followed by a subsequent persistent decrease. Maximal neutrophil gelatinase-associated lipocalin and calprotectin concentrations were 9-fold and 69-fold higher than their respective baseline values. The glomerular filtration rate was only transiently impaired at the first post-operative day after ischaemia/reperfusion injury (p = 0.049). Conclusion: Calprotectin and neutrophil gelatinase-associated lipocalin can be used to monitor clinical and sub-clinical tubular damage after nephron sparing surgery for kidney tumours. Urinary calprotectin concentrations start rising within 2 hours after ischaemia/reperfusion-induced kidney injury

Association between (A) the maximum urinary calprotectin increase and ischaemic time and (B) the maximum urinary NGAL increase and ischaemic time in Group 1 (NSS with renal ischaemia) in a linear regression model, as assessed by Spearman’s correlation test.

<p>Regression lines are plotted. Linear regression model: A: R = 0.16, B = 103.61 (95% CI -219.96–427.19), p = 0.51; B: R = 0.09, B = 1.71 (95% CI -7.88–11.31), p = 0.71. Spearman’s correlation: A: R = 0.29 (95% CI -0.19–0.66), p = 0.21; B: R = 0.26 (95% CI -0.22–0.64), p = 0.26. NSS, nephron sparing surgery for kidney tumours; B, regression co-efficient, slope of the regression line; R, correlation co-efficient; CI, confidence interval.</p

Concentrations of (A) urinary calprotectin, (B) urinary neutrophil gelatinase—associated lipocalin (NGAL), (C) plasma creatinine, and (D) the estimated glomerular filtration rate (eGFR) in patients undergoing nephron sparing surgery (NSS) for kidney tumours with renal pedicle clamping (renal ischaemia, Group 1) or without renal ischaemia (Group 2), and in patients undergoing radical nephrectomy for living kidney donation (Group 3).

<p>Data are presented as median and interquartile range. Asterisks indicate significant changes from baseline in a non-parametric analysis of variance (Kruskal-Wallis test, corrected for multiple comparisons by Dunn’s test as a post hoc pairwise multiple comparison). p.o., post-operative. * p < 0.05, ** p < 0.01, *** p < 0.001.</p