Pathophysiological implications of urinary peptides in hepatocellular carcinoma

SIMPLE SUMMARY: In this study, the application of capillary electrophoresis mass spectrometry enabled identification of 31 urinary peptides significantly associated with hepatocellular carcinoma diagnosis and prognosis. Further assessment of these peptides lead to prediction of cellular proteases involved in their development namely Meprin A subunit α and Kallikrein-6. Subsequent identification of the proteases was verified by immunohistochemistry in normal liver, cirrhosis and hepatocellular carcinoma. Histopathological assessment of the proteases revealed numerical gradient staining signifying their involvement in liver fibrosis and hepatocellular carcinoma formation. The discovered urinary peptides offered a potential noninvasive tool for diagnosis and prognosis of hepatocellular carcinoma. ABSTRACT: Hepatocellular carcinoma (HCC) is known to be associated with protein alterations and extracellular fibrous deposition. We investigated the urinary proteomic profiles of HCC patients in this prospective cross sectional multicentre study. 195 patients were recruited from the UK (Coventry) and Germany (Hannover) between 1 January 2013 and 30 June 2019. Out of these, 57 were HCC patients with a background of liver cirrhosis (LC) and 138 were non-HCC controls; 72 patients with LC, 57 with non-cirrhotic liver disease and 9 with normal liver function. Analysis of the urine samples was performed by capillary electrophoresis (CE) coupled to mass spectrometry (MS). Peptide sequences were obtained and 31 specific peptide markers for HCC were identified and further integrated into a multivariate classification model. The peptide model demonstrated 79.5% sensitivity and 85.1% specificity (95% CI: 0.81–0.93, p < 0.0001) for HCC and 4.1-fold increased risk of death (95% CI: 1.7–9.8, p = 0.0005). Proteases potentially involved in HCC progression were mapped to the N- and C-terminal sequence motifs of the CE-MS peptide markers. In silico protease prediction revealed that kallikrein-6 (KLK6) elicits increased activity, whilst Meprin A subunit α (MEP1A) has reduced activity in HCC compared to the controls. Tissue expression of KLK6 and MEP1A was subsequently verified by immunohistochemistry

Percutaneous transhepatic or endoscopic ultrasound-guided biliary drainage in malignant distal bile duct obstruction using a self-expanding metal stent: Study protocol for a prospective European multicenter trial (PUMa trial)

Background Endoscopic ultrasound-guided biliary drainage (EUS-BD) was associated with better clinical success and a lower rate of adverse events (AEs) than fluoroscopy-guided percutaneous transhepatic biliary drainage (PTBD) in recent single center studies with mainly retrospective design and small case numbers (< 50). The aim of this prospective European multicenter study is to compare both drainage procedures using ultrasound-guidance and primary metal stent implantation in patients with malignant distal bile duct obstruction (PUMa Trial). Methods The study is designed as a non-randomized, controlled, parallel group, non-inferiority trial. Each of the 16 study centers performs the procedure with the best local expertise (PTBD or EUS-BD). In PTBD, bile duct access is performed by ultrasound guidance. EUS-BD is performed as an endoscopic ultrasound (EUS)-guided hepaticogastrostomy (EUS-HGS), EUS-guided choledochoduodenostomy (EUS-CDS) or EUS-guided antegrade stenting (EUS-AGS). Insertion of a metal stent is intended in both procedures in the first session. Primary end point is technical success. Secondary end points are clinical success, duration pf procedure, AEs graded by severity, length of hospital stay, re-intervention rate and survival within 6 months. The target case number is 212 patients (12 calculated dropouts included). Discussion This study might help to clarify whether PTBD is non-inferior to EUS-BD concerning technical success, and whether one of both interventions is superior in terms of efficacy and safety in one or more secondary endpoints. Randomization is not provided as both procedures are rarely used after failed endoscopic biliary drainage and study centers usually prefer one of both procedures that they can perform best

Neuartige magnetokalorische Maschine für Kühl- und Heizanwendungen - MagKal : Schlussbericht ; Laufzeit des Vorhabens: 01.01.2012-30.06.2015

Aus dem Projektziel der Weiterführung von Erforschung und Anwendung des Magnetokalorischen Effekts (MCE) generierte sich für das IFW die praktische Demonstration der Vervielfachung sowie Umkehrung der Temperaturänderung. Grundlegend wurden Arbeiten zur Optimierung von Magnetfeld, Probenform und Wärmetausch zwischen MCE-Material und Fluid vorangestellt. Nach umfangreichen theoretischen und kalkulatorischen Betrachtungen zum Magnetfeld, flossen diese Erkenntnisse direkt in die Entwicklung eines Zyklensimulators ein. Durch diese eigenerrichtete Versuchsanlage konnte das Magnetkonzept für den Demonstrator bestätigt und optimiert werden. Neben dem quasi Referenzmaterial Gadolinium wurde mit apparativer Unterstützung aus dem IFW Lanthan-Eisen-Silizium-Proben gefertigt. In nachfolgenden Untersuchungen wurden umfangreiche Testreihen zum effizienten Wärmetausch vom MCE-Material an das geeignetste Übertragungsfluid und zur bestmöglichen Probengeometrie absolviert. Die apparative Grundlage dazu stellt die thermische Testanlage des IFW dar. Eine Steigerung des technischen Aufwandes gegenüber dem Zyklierer zeigt sich in umfangreicher Sensorik und einem eigenen Messprogramm. Durch realitätsnahe Versuche im Feld eines starken Elektromagneten wurden wichtig Erkenntnisse für die Gestaltung des Probenmoduls im Demonstrator gewonnen. Der im IFW Dresden in Betrieb genommene MagKal-Demonstrator ermöglicht eine Vielzahl an Variationen von Parametern des magnetokalorischen Prozesses. Beispielgebend seien hier Durchfluss, Öffnungsverschiebung, Taktzeit, … genannt. Dieses Konzept erlaubt neben der Erweiterungsperspektive die Option für seriellen bzw. parallelen Betrieb der Module. Die Probenkammern der Module erlauben eine Untersuchung verschiedener Probengeometrien und /oder –materialien. Darüber hinaus können für diesen Demonstrator alle Parameter im Kühl- und Wärmepumpenbetrieb gefahren werden. Somit verschafft die Breite der experimentell beeinflussbaren Versuchsparameter dem Demonstrator einen unikalen Status. Diese Eigenschaften sind die wesentlichen Alleinstellungsmerkmale des im MagKal-Projekt entwickelten Demonstrators. Vielversprechende Ansätze zur Anwendung des erstellten Demonstrators gibt es für die ökologisch und energetisch optimierte technische Gebäudeklimatisierung, da in dieser Leistungsklasse vergleichsweise langsame Prozesse mit geringerer Temperaturdifferenz stattfinden. Es wird erwartet, dass es auf der Grundlage der Projektergebnisse gelingt, industrielle Kooperationspartner zur Weiterentwicklung in Richtung kommerziell verwertbarer Produkte zu gewinnen.From the project aim of continuing science and application of the Magneto Caloric Effect (MCE) arose for the IFW the practical demonstration of multiplying as well as inversion of temperature differences. Fundamental studies concerning magnetic field optimization, sample shapes and heat transport from MCE-material to fluid are done previously. After extensive theoretical and calculative dimensioning of the magnetic field, these new knowledges flu directly into the development of a cycle simulator. This self-made testing plant was well suitable to confirm and optimize the magnetic setup for the demonstrator. Beside of preparing the reference material gadolinium samples, new lanthanum-iron-silicon-samples were made with the instrumental help of the IFW. In the following investigations at the IFW, manifold series of tests concerning the effective heat transport from the MCE-Material to the best suitable transfer fluid and the best sample geometry were passed. The instrumental basis for these tests is the thermal test bench at the IFW. Comparing to the cycle test rig, comprehensive sensor devices and a suitable measuring software were implemented at the thermal test bench. Main results for the most promising modular concept of the demonstrator came up with the close-to-reality-experiments in the magnetic field of a strong electromagnet. The specially built demonstrator at the IFW allows wide variations of process parameters. For example flow rate, valve opening time shift, cycle time. This concept provides options for increasing the number of modules as well as operating the modules in serial or parallel order. The universal sample cells enables the intensive research of different sample geometries and/ or materials. Above that, all parameters for this demonstrators can be run in refrigeration and heat pump operation. This setup with its wide variety of parameters enforces the demonstrator to get a unique state. This features are the main stand-alone-properties of the developed demonstrator within the MagKal-project. Promising ideas for application of the demonstrator design can be found at the ecological and economical friendly air-conditioning of buildings, because just small temperature differences and slow processes occurs in that performance category. It is expected that based on the project results industrial partners will be attracted for further development of commercially exploitable products

When a respiratory pathogen turns to the skin: cutaneous tuberculosis in a lung transplant patient

A 62-year-old male, who received immune suppressive therapy due to a lung transplantation several years ago, developed multiple painful abscesses in the right forearm. First misdiagnosed as staphylococcal abscesses, Mycobacterium tuberculosis was eventually cultured from the abscesses. In addition, the patient also suffered from pulmonary tuberculosis and respiratory specimens were also culture-positive for Mycobacterium tuberculosis . Cutaneous tuberculosis must be kept as a differential diagnosis in the case of abscess-like lesions on the skin, especially in immunocompromised patients. Mycobacteria specific tests (polymerase chain reaction in respiratory samples and wound smears) and antituberculotic combination therapy are necessary to treat Mycobacterium tuberculosis infection/reactivation adequately

Angiopoietin-2 and biliary diseases: elevated serum, but not bile levels are associated with cholangiocarcinoma.

BACKGROUND:The diagnosis of cholangiocarcinoma (CC) is challenging especially in patients with primary sclerosing cholangitis (PSC) and often delayed due to the lack of reliable markers. Angiopoietin-2 (Angpt-2) has been employed as a biomarker of angiogenesis and might be involved in tumor neoangiogenesis. AIM:To evaluate the diagnostic potential of Angpt-2 as a biomarker to detect patients with CC. METHODS:Bile and serum Angpt-2 levels were measured in patients with CC (n=45), PSC (n=74), CC complicating PSC (CC/PSC) (n=11) and patients with bile duct stones (n=37) in a cross sectional study. Diagnostic accuracy of Angpt-2 was compared to carbohydrate antigen 19-9 (CA19-9). Fluorescent immunohistochemistry from human CC liver tissue samples was performed to localize the origin of Angpt-2. RESULTS:Serum Angpt-2 concentration was significantly elevated in patients with CC compared to control patients (p<0.05). Diagnostic accuracy of Angpt-2 as determined by receiver operating characteristic (ROC) analysis resulted in a higher area under the curve (AUC) value compared to CA19-9 (AUC: 0.85 versus 0.77; 95% confidence interval (CI): 0.74-0.93 versus 0.65-0.87, respectively). Angpt-2 was also detectable in bile, but was not associated with the presence of CC. Immunohistochemistry revealed a strong induction of Angpt-2 expression in the tumor vasculature. CONCLUSIONS:Circulating Angpt-2 in serum might be a promising protein candidate locally derived from the tumor vasculature in patients with CC. Measurement of Angpt-2 in serum may be useful for diagnosis and further clinical management of patients with CC

MicroRNAs in Serum and Bile of Patients with Primary Sclerosing Cholangitis and/or Cholangiocarcinoma.

BACKGROUND AND AIM:Patients with primary sclerosing cholangitis (PSC) are at high risk for the development of cholangiocarcinoma (CC). Analysis of micro ribonucleic acid (MiRNA) patterns is an evolving research field in biliary pathophysiology with potential value in diagnosis and therapy. Our aim was to evaluate miRNA patterns in serum and bile of patients with PSC and/or CC. METHODS:Serum and bile from consecutive patients with PSC (n = 40 (serum), n = 52 (bile)), CC (n = 31 (serum), n = 19 (bile)) and patients with CC complicating PSC (PSC/CC) (n = 12 (bile)) were analyzed in a cross-sectional study between 2009 and 2012. As additional control serum samples from healthy individuals were analyzed (n = 12). The miRNA levels in serum and bile were determined with global miRNA profiling and subsequent miRNA-specific polymerase chain reaction-mediated validation. RESULTS:Serum analysis revealed significant differences for miR-1281 (p = 0.001), miR-126 (p = 0.001), miR-26a (p = 0.001), miR-30b (p = 0.001) and miR-122 (p = 0.034) between patients with PSC and patients with CC. All validated miRNAs were significantly lower in healthy individuals. MiR-412 (p = 0.001), miR-640 (p = 0.001), miR-1537 (p = 0.003) and miR-3189 (p = 0.001) were significantly different between patients with PSC and PSC/CC in bile. CONCLUSIONS:Patients with PSC and/or CC have distinct miRNA profiles in serum and bile. Furthermore, miRNA concentrations are different in bile of patients with CC on top of PSC indicating the potential diagnostic value of these miRNAs