Preoperative ultrasound elastography for postoperative pancreatic fistula prediction after pancreatoduodenectomy: A prospective study.

BACKGROUND: Postoperative pancreatic fistulas are the most frequent major complications after pancreatoduodenectomy. The soft pancreatic texture is a critical, independent risk factor for postoperative pancreatic fistulas after pancreatoduodenectomy. The current gold standard for postoperative pancreatic fistula risk evaluation consists of the surgeon's intraoperative palpation of the pancreatic texture and, thus, lacks objectivity. In this prospective study, we used ultrasound-based shear-wave elastography, image data analysis, and a fistula risk score calculator to correlate the stiffness of pancreatic tissue with the occurrence of clinically relevant postoperative pancreatic fistulas. METHODS: We included 100 patients with pancreatic pathologies (71% pancreatic ductal adenocarcinoma) and 100 healthy individuals who were preoperatively assessed via real-time tissue ultrasound-based shear-wave elastography on a Philips EPIQ 7 ultrasound device and had pancreatic parenchyma histologically evaluated with manually stained images. RESULTS: We found a significant difference in the mean elasticity between the soft (1.22 m/s) and the hard pancreas group (2.10 m/s; P < .0001). The mean elasticity significantly correlated with the pancreatic fibrosis rate and the appearance of a postoperative pancreatic fistula after pancreatoduodenectomy. Low elasticity (≤1.2 m/s, mean) correlated with soft and high elasticity (>2.0 m/s, mean) with hard pancreatic parenchyma, as assessed by pathologic evaluation. Multivariate analysis revealed a mean elasticity of <1.3 m/s as a significant cut-off predictor for clinically relevant postoperative pancreatic fistulas (P = .003; Youden-Index = 0.6945). CONCLUSION: Preoperative ultrasound-based shear-wave elastography is a feasible and objective clinical diagnostic modality in evaluating pancreatic tissue stiffness. A mean pancreatic elasticity of <1.3 m/s was a significant independent risk predictor of clinically relevant postoperative pancreatic fistulas after pancreatoduodenectomy.

TPX2 expression as a negative predictor of gemcitabine efficacy in pancreatic cancer.

BACKGROUND: Targeting protein for Xenopus kinesin-like protein 2 (TPX2) overexpression in human tumours is associated with increased malignancy. Its effect on gemcitabine resistance in pancreatic ductal adenocarcinoma (PDAC) has not been studied yet. METHODS: The prognostic impact of TPX2 expression was examined in the tumour tissue of 139 patients with advanced PDAC (aPDAC) treated within the AIO-PK0104 trial or translational trials and of 400 resected PDAC (rPDAC) patients. The findings were validated using RNAseq data of 149 resected PDAC patients. RESULTS: In the aPDAC cohorts, 13.7% of all samples showed high TPX2 expression, conferring significantly shorter progression-free survival (PFS, HR 5.25, P < 0.001) and overall survival times (OS, HR 4.36, P < 0.001) restricted to gemcitabine-based treated patients (n = 99). In the rPDAC cohort, 14.5% of all samples showed high TPX2 expression, conferring significantly shorter disease-free survival times (DFS, HR 2.56, P < 0.001) and OS times (HR 1.56, P = 0.04) restricted to patients treated with adjuvant gemcitabine. RNAseq data from the validation cohort confirmed the findings. CONCLUSIONS: High TPX2 expression may serve as a negative predictor of gemcitabine-based palliative and adjuvant chemotherapy in PDAC and could be used to inform clinical therapy decisions. CLINICAL TRIAL REGISTRY: The clinical trial registry identifier is NCT00440167.

Serum biomarker panel diagnostics in pancreatic ductal adenocarcinoma: the clinical utility of soluble interleukins, IFN-γ, TNF-α and PD-1/PD-L1 in comparison to established serum tumor markers.

PURPOSE: Novel biomarkers to better predict outcome and select the best therapeutic strategy for the individual patient are necessary for pancreatic ductal adenocarcinoma (PDAC). METHODS: Using a panel assay, multiple biomarkers (IFN-γ, IL-10, IL-6, IL-8, TNF-α, CEA, CA 19-9, CYFRA 21-1, HE4, PD-1 and PD-L1 levels) were measured in serum samples of 162 patients with resected, locally advanced and metastatic PDAC in this retrospective single-center study. Optimal cut-off values to differentiate prognostic subgroups with significantly different overall survival (OS) were determined by receiver operator characteristics and Youden Index analysis. Marker levels were assessed before the start of chemotherapy and correlated with OS by univariate and multivariate Cox analysis. RESULTS: Median OS for resected patients was 28.2 months, for locally advanced patients 17.9 months and for patients with metastatic disease 8.6 months. CYFRA 21-1 and IL-8 discriminated metastatic from locally advanced patients best (AUC 0.85 and AUC 0.81, respectively). In univariate analyses, multiple markers showed prognostic relevance in the various subgroups. However, multivariate Cox models comprised only CYFRA 21-1 in the resected group (HR 1.37, p = 0.015), IL-10 in locally advanced PDAC (HR 10.01, p = 0.014), as well as CYFRA 21-1 and CA 19-9 in metastatic PDAC (p = 0.008 and p = 0.010) as an independent prognostic marker for overall survival. CONCLUSION: IL-10 levels may have independent prognostic value in locally advanced PDAC, whereas CYFRA 21-1 levels are prognostic after PDAC surgery. CYFRA 21-1 and IL-8 have been identified to best discriminate metastatic from locally advanced patients.

Bacterial Lipopolysaccharide as a Negative Predictor of Adjuvant Gemcitabine Efficacy in Pancreatic Cancer.

Adjuvant gemcitabine (aGC) is one standard of care after pancreatic ductal adenocarcinoma (PDAC) resection. No biomarker for its efficacy is established. As bacteria mediate gemcitabine resistance, we analyzed whether lipopolysaccharide (LPS) as surrogate for bacterial colonization is prognostic in PDAC patients treated with aGC or without aGC adjuvant gemcitabine. We detected LPS in 86 tumors from 376 patients, which defined a specific microbiome as revealed by 16 s-rRNA-sequencing. In the 230 aGC patients, LPS conferred worse disease-free survival (8.3 vs 13.7 months; hazard ratio = 1.75, 95% confidence interval = 1.22 to 2.49; log-rank P = .002) and overall survival (21.7 vs 28.5 months; hazard ratio = 1.80, 95% confidence interval = 1.23 to 2.57; log-rank P = .001) but not in the 146 naGC patients, which was confirmed in an independent validation cohort (n = 178). LPS may serve as a negative predictor for aGC efficacy in PDAC, which suggests a role for microbiome modification to overcome bacteria-mediated chemotherapy resistance.

The impact of adjuvant therapy on outcome in UICC stage I pancreatic cancer.

Adjuvant chemotherapy has become standard of care for pancreatic ductal adenocarcinoma (PDAC) as it improves patient outcome. However, its clinical meaning in early-stage, UICC I tumors remains uncertain. We examined the effect of adjuvant therapy on disease-free survival (DFS) and overall survival (OS) of UICC stage I PDAC patients treated at an academic tertiary care center between 2000 and 2016. Among 124 patients (69 male, 55 female; median age 68 years, range 41-84 years) with UICC stage I disease, adjuvant therapy improved both DFS (19.8 vs 12.8 months, HR 0.59, 95% CI: 0.37-0.94, P = .03) and OS (40.9 vs 20.3 months, HR 0.54, 95% CI: 0.35-0.84, P = .005). Multivariate analyses and propensity score matching confirmed the prognostic impact of adjuvant therapy independent of localization, differentiation and R-status. Thus, every patient with UICC I PDAC should receive adjuvant chemotherapy as it may improve outcome significantly. Our findings support the concept of PDAC as systemic disease from early stages on.

Bacterial lipopolysaccharide as negative predictor of gemcitabine efficacy in advanced pancreatic cancer - translational results from the AIO-PK0104 Phase 3 study.

BACKGROUND: Gram-negative bacteria mediated gemcitabine resistance in pre-clinical models. We determined if intratumoural lipopolysaccharide (LPS) detection by immunohistochemistry is associated with outcome in advanced pancreatic ductal adenocarcinoma (PDAC) treated with gemcitabine and non-gemcitabine containing 1st-line chemotherapy. METHODS: We examined LPS on tumour tissue from 130 patients treated within the randomised AIO-PK0104 trial and a validation cohort (n = 113) and analysed the association of LPS detection to patient outcome according to treatment subgroups. RESULTS: In 24% of samples from the AIO-PK0104 study LPS was detected; in LPS-positive patients median OS was 4.4 months, compared to 7.3 months with LPS negative tumours (HR 1.732, p = 0.010). A difference in OS was detected in 1st-line gemcitabine-treated patients (n = 71; HR 2.377, p = 0.002), but not in the non-gemcitabine treatment subgroup (n = 59; HR 1.275, p = 0.478). Within the validation cohort, the LPS positivity rate was 23%, and LPS detection was correlated with impaired OS in the gemcitabine subgroup (n = 94; HR 1.993, p = 0.008) whereas no difference in OS was observed in the non-gemcitabine subgroup (n = 19; HR 2.596, p = 0.219). CONCLUSIONS: The detection of intratumoural LPS as surrogate marker for gram-negative bacterial colonisation may serve as a negative predictor for gemcitabine efficacy in advanced PDAC. CLINICAL TRIAL REGISTRY: The Clinical trial registry identifier is NCT00440167.

POLE gene hotspot mutations in advanced pancreatic cancer.

PURPOSE: Pancreatic ductal adenocarcinoma (PDAC) is a devastating disease, lacking relevant prognostic and predictive biomarkers. DNA polymerase epsilon (POLE) has important functions in the maintenance of genetic stability during DNA replication and has previously been associated with favorable prognosis in endometrial and colorectal cancer. However, its relevance in advanced pancreatic cancer (aPDAC) has not been examined to date. METHODS: Using pyrosequencing on tumoral DNA extracted from 60 samples from the AIO-PK0104 study as well as 55 samples from completed translational trials, we examined POLE hotspot mutations in exon 9 (P286R) and exon 13 (V411R/L/M) in the POLE gene exonuclease domain. DNA extracted from 37 endometrial carcinomas were tested as positive controls. Publically available sequencing databases were searched for POLE mutations in PDAC samples. RESULTS: Fifty-three patients (pts) were men, 62 pts were women, median age was 61.2 years. Median overall survival (OS) was 7.4 months and median progression free survival (PFS) was 4.0 months. In four of the 37 endometrial carcinomas POLE mutations were detected in exon 9 (10.8%) and none in exon 13. In none of the overall 115 aPDAC tumors POLE gene hotspot mutations could be detected. CONCLUSION: Mutations in the hotspot regions of exon 9 and 13 of the POLE gene are very rare events in advanced pancreatic cancer. Thus, it is unlikely that POLE gene mutations contribute to genetic instability in the vast majority of aPDAC. POLE mutation does not serve as a relevant biomarker and should not be tested on a regular basis in PDAC.

Acute ischaemic stroke and myocardial infarction after chemotherapy with vinorelbine for non-small cell lung cancer: a case report.

Vinorelbine is an important chemotherapeutic agent which is used in metastatic non-small cell lung cancer. Case reports have described the occurrence of acute cardiac ischaemic events as a side effect. It has not been established whether the suspected mechanisms for cardiac ischaemia might also cause other vascular events. We report about a 70-year-old male with metastatic non-small cell lung cancer who received vinorelbine as an outpatient. The patient presents with a cardiovascular risk profile. He was admitted to the hospital 3 days later with acute left-sided hemiplegia and hemianopia. Brain computed tomography (CT) demonstrated acute right hemispheric ischaemic stroke. Nine days after admission, the patient additionally suffered ST elevation myocardial infarction. A coronary angiogram demonstrated high grade stenosis of the right coronary artery treated with two bare-metal stents. Caution should be noted in patients who present with a cardiovascular risk profile as they might be vulnerable experiencing acute ischaemic events.

pH-Driven Self-Assembly of Acidic Microbial Glycolipids.

Microbial glycolipids are a class of well-known compounds, but their self-assembly behavior is still not well understood. While the free carboxylic acid end group makes some of them interesting stimuli-responsive compounds, the sugar hydrophilic group and the nature of the fatty acid chain make the understanding of their self-assembly behavior in water not easy and highly unpredictable. Using cryo-transmission electron microscopy (cryo-TEM) and both pH-dependent in situ and ex situ small angle X-ray scattering (SAXS), we demonstrate that the aqueous self-assembly at room temperature (RT) of a family of ß-d-glucose microbial glycolipids bearing a saturated and monounsaturated C18 fatty acid chain cannot be explained on the simple basis of the well-known packing parameter. Using the "pH-jump" process, we find that the molecules bearing a monosaturated fatty acid forms vesicles below pH 6.2, as expected, but the derivative with a saturated fatty acid forms infinite bilayer sheets below pH 7.8, instead of vesicles. We show that this behavior can be explained on the different bilayer membrane elasticity as a function of temperature. Membranes are either flexible or stiff for experiments performed at a temperature respectively above or below the typical melting point, TM, of the lipidic part of each compound. Finally, we also show that the disaccharide-containing acidic cellobioselipid forms a majority of chiral fibers, instead of the expected micelles.

Improving Intensive Care Unit and Ward Utilization by Adapting Master Surgery Schedules.

With increasing organizational and financial pressure on hospitals, each individual surgical treatment has to be reviewed and planned thoroughly. Apart from the expensive operating room facilities, proper staffing and planning of downstream units, like the wards or the intensive care units (ICUs), should be considered as well. In this article, we outline the relationship between a master surgery schedule (MSS), i.e., the assignment of surgical blocks to medical specialties, and the bed demand in the downstream units using an analytical model. By using historical data retrieved from the clinical information system and a patient flow model, we applied a recently developed algorithm for predicting bed demand based on the MSSs for patients of 3 surgical subspecialties of a hospital. Simulations with 3 different MSSs were performed. The impact on the required amount of beds in the downstream units was analyzed. We show the potential improvements of the current MSS considering 2 main goals: leveling workload among days and reduction of weekend utilization. We discuss 2 different MSSs, one decreasing the weekend ICU utilization by 20% and the other one reducing maximum ward bed demand by 7%. A test with 12 months of real-life data validates the results. The application of the algorithm provides detailed insights for the hospital into the impact of MSS designs on the bed demand in downstream units. It allowed creating MSSs that avoid peaks in bed demand and high weekend occupancy levels in the ICU and the ward.

Target temperature management of 33 degrees C exerts beneficial haemodynamic effects after out-of-hospital cardiac arrest.

BACKGROUND: Accumulating evidence indicates that target temperature management (TTM) is beneficial in patients resuscitated after cardiac arrest since it appears to improve neurological outcome. However, the optimal cooling method (surface vs. intravascular) has not yet been specified. Substantial heart disease is present in most of these patients and therefore haemodynamic effects of cooling need to be considered very carefully. We analysed the haemodynamic response to TTM in patients treated with surface versus intravascular cooling following out-of-hospital cardiac arrest. METHODS AND RESULTS: In this observational study 63 consecutive subjects presenting to the hospital after successful resuscitation following of out-of-hospital cardiac arrest received an intravascular (40 patients) or external cooling device (23 patients) to induce TTM. While with intravascular cooling the target temperature of 33 degrees C was reached after 159 minutes, the minimum temperature achieved with surface cooling was about 35 degrees C after 437 minutes. Haemodynamic parameters were recorded in a 4-hour rhythm for the first 12 hours after induction of hypothermia. Generally, TTM of 33 degrees C resulted in a higher systemic vascular resistance index (749 vs. 467 dyn*sec/cms/m2; P= 0.04) but also in a marked reduction of heart rate (67.70 vs. 100.00 bpm; P < 0.001), a higher mixed venous oxygen saturation (76 vs. 68%; P = 0.016), and a higher stroke volume index (45 vs. 33 mI/m2; P = 0.036). TTM additionally resulted in a higher cardiac power index (0.55 vs. 0.46 Watt/m2; P = 0.024). CONCLUSION: TTM of 33 degrees C compared to 35 degrees C exerts beneficial haemodynamic effects and might be viewed as an adjunct inotropic therapy avoiding the undesired side effects of vasoactive substances.

Genome Sequence of the Basidiomycetous Fungus Pseudozyma aphidis DSM70725, an Efficient Producer of Biosurfactant Mannosylerythritol Lipids.

Pseudozyma aphidis is an efficient producer of mannosylerythritol lipids exceeding concentrations of >100 g/liter from renewable feed stocks. Additionally, a biosurfactant cellobiose lipid is also secreted during nitrogen limitation. Here, we describe the sequencing of P. aphidis to unravel the genomic basis of biosurfactant metabolism in P. aphidis.

Pre-hospital discharge testing of defibrillator without anesthesia: effects on psychological symptoms during clinical follow-up.

OBJECTIVE: After the implantation of an implantable cardioverter-defibrillator (ICD), patients often fear therapeutic shock. The extent to which the experience of pre-hospital discharge (PHD) testing without anesthesia after ICD implantation, under observation by a physician, affects shock-related anxiety symptoms on follow-up has not been investigated as yet. METHODS: In a prospective, randomized controlled trial, 44 patients with a primary prevention indication for an ICD were randomly assigned to experience PHD testing without anesthesia (n = 23) or with anesthesia (n = 21). Patients were longitudinally evaluated before (T(1)), shortly after (T(2)), and 3 months after (T(3)) PHD testing. During the respective PHD testings, the course of patients' serum cortisol release was measured. RESULTS: During PHD testing, patients without anesthesia showed a significantly higher serum cortisol release than patients with anesthesia (F(4,152) = 22.227, p < .001). Patients who experienced PHD testing without anesthesia felt significantly safer with the ICD (U = 165.000, p = .040), would significantly more often recommend other patients to undergo PHD testing without anesthesia (χ(2) = 12.013, p = .002), and showed significantly lower levels of general shock-related anxiety shortly afterward (F(1,42) = 6.327, p = .02) and 3 months after PHD testing (F(1,41) = 8.603, p = .005). CONCLUSIONS: The implementation of PHD testing without anesthesia is associated with lower anxiety concerning therapeutic shock. Patients should be advised about the effects of PHD testing without anesthesia on their psychological well-being in the long run.

Integrating strength tests of amputees within the protocol of conventional clinical gait analysis: a novel approach.

Clinical experience tells us that the lower-limb amputees are one of the patient groups who clearly suffer from a strength deficit in their involved side. However, there is no obvious evidence for the relation between the residual limb strength and walking ability in this population. Correlating the results of the conventional clinical gait analysis (CGA) with strength tests could help to find out how deficits in strength impact the amputees' gait. In this contribution, a new device for measuring the isometric muscle strength of the hip and the knee was tested for feasibility. Three groups were tested: one group of 11 healthy subjects (29±5 years) to test the repeatability of the device, two unilateral amputees (one transfemoral for 56 years, one transtibial for 65 years), and a reference group of 17 healthy subjects (55±10 years). The new method presents an adequate technique to integrate strength testing within a standard protocol of the CGA. Results showed to be repeatable within sessions [i.e., within-day, intraclass correlation coefficient (ICC)>0.972] and between repeated measurements (i.e., day-to-day, ICC>0.765). The tested amputees showed clear deficits in maximum isometric joint moments in their most distal joint. The first results suggest evidence for a relation between the maximum isometric joint moments and gait deviations in amputees.

Endothelial differentiation of adipose-derived mesenchymal stem cells is improved by epigenetic modifying drug BIX-01294.

Chromatin remodeling plays an essential role in regulation of gene transcription. Consequently, targeted changes in chromatin may also augment pluripotency of somatic cells. The aim of the present study was to evaluate the effect of epigenetic drug BIX-01294 (BIX), a histone G9a inhibitor, on DNA methylation, expression of pluripotency genes POU5F1 (isoform a), NANOG, KLF4, and CMYC in mesenchymal stem cells, and the ability to increase their differentiation potential into endothelial cells (ECs). Human adipose-derived mesenchymal stem cells (AdMSCs) were isolated from abdominal adipose tissue. Cells were pre-treated with BIX for 48h and further differentiated in endothelial medium for 7 and 14 days. Global DNA methylation was determined by MethyLight application, expression of genes for pluripotency, endothelial and angiogenic markers by SYBRGreen-based real-time PCR, immunocytochemistry, and immunobloting. Following treatment with BIX, DNA methylation status of AdMSCs was significantly reduced by 53% (p=0.008), the expression of POU5F1 and NANOG was increased by 2.2-fold (p=0.016) and 1.5-fold (p<0.001), respectively. Furthermore, BIX pre-treatment improved the differentiation capacity of AdMSCs into ECs and significantly increased expression of several endothelial markers and factors involved in blood vessel formation: VCAM-1, PECAM-1, von Willebrand factor, VEGFR-2, PDGF, and ANG-1 in comparison with AdMSCs without BIX pre-treatment. In the present study we demonstrate that epigenetic modifying drug BIX-01294 is able to increase the ability of AdMSCs to differentiate into ECs engaging DNA and histone methylation. Hence, BIX-01294 might serve as a simple tool to increase the differentiation potential of AdMSCs.