Cornelia M. Gelbmann

Differential Activation of Cytokine Secretion in Primary Human Colonic Fibroblast/Myofibroblast Cultures

BACKGROUNDnFibroblasts and myofibroblasts are known to secrete a wide spectrum of cytokines, but the individual spectrum is tissue-specific. We investigated the effect of cell activation on cytokine secretion of isolated human colonic fibroblasts/myofibroblasts from control patients and patients with mucosal inflammation.nnnMETHODSnPrimary cultures of human colonic submucosal fibroblasts/myofibroblasts were incubated with IL-1alpha (100 U/ml), IL-Ibeta (10 ng/ml), IL-10 (10 ng/ml), TNF (10 ng/ml), PMA (10 ng/ml), LPS (50 ng/ml), IL-4 (10 ng/ml), or a combination of IL-1 and TNF. Secreted cytokines were determined by ELISA. NF-kappaB activation was demonstrated by electrophoretic mobility-shift assays (EMSA).nnnRESULTSnIncubation of colonic fibroblasts/myofibroblasts with IL-1, LPS, TNF and PMA induced secretion of IL-6, IL-8, M-CSF and GM-CSF. IL-8 and IL-6 secretion could be stimulated by IL-1alpha, IL-1beta, TNF, PMA and LPS within 6 h of incubation. IL-6 secretion was stimulated from 0.5 +/- 0.01 pg/h x microg fibroblast protein to 18.5 +/- 2.6 pg/h x microg fibroblast protein with IL-1beta (P < 0.01). IL-8 secretion was stimulated from 1.0 +/- 0.1 pg/h x microg fibroblast protein to 41.1 +/- 3.6 pg/h x microg (P < 0.005). IL-4 and IL-10 did not change cytokine secretion significantly. No significant differences between cultures from normal and inflamed mucosa were observed. TNF and IL-1 induced NF-kappaB activation. ALLN, a proteasome and NF-kappaB activation inhibitor, reduced TNF-mediated IL-8, GM-CSF and M-CSF induction significantly, whereas induction of IL-6 secretion remained unchanged.nnnCONCLUSIONnHuman colonic myofibroblasts can secrete large amounts of IL-6, IL-8, M-CSF and GM-CSF upon stimulation. The induction of IL-8, M-CSF and GM-CSF, but not of IL-6 secretion, is mediated mainly by NF-kappaB activation. The cytokine profile and the total amounts of cytokines released suggest that colonic myofibroblasts can play a role in leukocyte recruitment and during mucosal inflammation. They therefore have to be regarded as an important part of the mucosal immune system.

Autocrine Fibronectin-Induced Migration of Human Colonic Fibroblasts

OBJECTIVES:A central event during wound repair is the migration of activated fibroblasts to the wound area. Thus far, the mechanisms inducing migration of colonic lamina propria fibroblasts (CLPF) have not been studied in detail. Previously, we have shown that CLPF secrete factors that are essential to their ability to migrate in response to different growth factors.METHODS:Primary human CLPF were obtained from endoscopic biopsies or surgical specimens taken from normal mucosa areas of patients undergoing surveillance colonoscopy or surgery for colorectal carcinoma. Migration assays of CLPF were performed in the modified 48-well Boyden chamber.RESULTS:Conditioned medium of CLPF collected after 24-h stimulated migration of CLPF (22 ± 2 cells/hpf). Filtration of conditioned medium through a 300-kDa filter reduced the migration-inducing potential in subsequent migration assays to 2 ± 1 cells/hpf, filtration through a 100-kDa filter abolished migration of CLPF completely, indicating that large molecules such as extracellular matrix components could be responsible for the induction of CLPF migration. Enzyme-linked immunosorbent assays revealed the presence of fibronectin in conditioned medium (17.3 μg/ml). Immunoprecipitation of fibronectin in conditioned medium of CLPF reduced the migration-inducing potential by 63%. Addition of fibronectin to fibronectin-depleted conditioned medium reconstituted the migration. Dose-response assays with fibronectin (1–100 μg/ml) diluted in nonconditioned medium induced migration of CLPF in a dose-dependent manner. Maximum migration was induced with 25 μg/ml fibronectin (37 ± 5 cells/hpf).CONCLUSIONS:Fibronectin is an autocrine and paracrine factor essential for intestinal fibroblast migration. Fibronectin induces migration of intestinal fibroblasts and is essential for their ability to migrate in response to different growth factors. A detailed understanding of the regulation of the migration of intestinal fibroblasts is necessary to gain further insights in the pathophysiology of stricture and fistula formation.

Outcome of patients with ischemic-like cholangiopathy with secondary sclerosing cholangitis after liver transplantation

Abstract Background and aims. Sclerosing cholangitis in critically ill patients (SC-CIP) with sepsis and acute respiratory distress syndrome (ARDS) is a cholestatic liver disease with a rapid progression to liver cirrhosis and hepatic failure. Data on outcome of these patients after liver transplantation (LT) are sparse. Patients and methods. Eleven patients (46 ± 12 years; mean labMELD-score: 27 ± 7) with SC-CIP underwent LT. Six patients had severe polytrauma with multiple bone fractures, sepsis and ARDS. Five non-traumatic patients acquired SC-CIP during long-term intensive-care-unit stays due to sepsis and ARDS. Time to diagnosis, the microbiologic results and the survival rates after LT were evaluated. Results. SC-CIP was diagnosed by endoscopic retrograde cholangiopancreatography (ERCP) within 3 ± 1 months after manifestation of cholestasis and histologically confirmed in explanted livers. The predominant microorganisms isolated in bile were: Enterococcus and Candida albicans. Mean follow-up after LT was 28 ± 20 months. One female patient (non-traumatic) died due to sepsis 26 days after LT. All other patients left the hospital alive, but two (non-traumatic) patients died from sepsis, and one (traumatic) patient died in a hemorrhagic shock, thereafter. Seven of 11 patients (5 with polytrauma) are still alive and have a good quality of life. The survival of the SC-CIP patients after LT was comparable with that of patients transplanted due to alcoholic liver cirrhosis. Conclusion. SC-CIP develops rapidly within several months. Enterococcus and C. albicans were the main isolated microorganisms in the bile. Sepsis was the main cause of death after LT. Overall, SC-CIP is a good indication for LT in selected patients.

Causes of bolus impaction in the esophagus.

BackgroundBolus impaction in the esophagus is a common indication for emergency endoscopy. The aim of this study was to determine the most common causes of esophageal bolus impaction.MethodsIn this retrospective study, data of 54 patients (41 male, 13 female) with bolus impaction in the esophagus were analyzed. Type and localization of the bolus and the endoscopic extraction tool used were evaluated. In 48 of 54 patients (89%), biopsy samples were taken of the esophagus for histological examination.ResultsMean age of the patients was 53xa0±xa020xa0years. Fourteen of 54 patients (26%) had experienced bolus impaction previously. Meat bolus (nxa0=xa035, 65%) was the most common cause of esophageal obstruction. In most cases, boluses were found in either the distal (nxa0=xa031) or the proximal (nxa0=xa018) esophagus. In 22 patients (41%), the bolus was pushed into the stomach by the endoscope. In most other cases the bolus, including foreign bodies, could be removed with the 5-arm polyp grasper or alligator forceps. Main causes of bolus impaction were eosinophilic esophagitis (nxa0=xa010) or reflux disease with or without peptic stenosis (nxa0=xa010), respectively.ConclusionBolus impaction is frequently correlated with eosinophilic esophagitis and reflux esophagitis; therefore, diagnostic workup should include esophageal biopsy sampling.

Qualitative and quantitative clinical evaluation of the laser-assisted ratio analyser for detection of Helicobacter pylori infection by 13C-urea breath tests

Objectives Currently, the 13C-urea breath test is the reference method for non-invasive diagnosis of Helicobacter pylori infection and therapy control. Therefore, new technologies have been developed to measure the ratio of 13CO2/12CO2 in breath. The laser-assisted ratio analyser (LARA) device is based on optogalvanic effects of the stimulated CO2 molecules. Design In this study, the LARA system is prospectively compared to conventional isotope ratio mass spectrometry (IRMS) analysis of 13C-urea breath tests. Methods The 13C-urea breath test was used to screen 103 patients for H. pylori infection. Breath samples were analysed by LARA and IRMS techniques. Results Seven breath tests could not be analysed by the LARA system, one by IRMS. Out of the remaining 95 breath tests, 13 were positive for H. pylori infection (13.7%). In reference to IRMS analysis (with a cut-off of > 5 δ‰ at 30 min), LARA produced one false positive and one false negative breath test result giving a sensitivity of 92.3% and a specificity of 98.8%. The mean difference in delta over baseline values between IRMS and LARA measurements was 2.02 δ‰ ± 5.48 δ‰. Conclusion LARA allows the reliable qualitative evaluation of 13C-urea breath tests, but the quantitative results differ from IRMS findings.

Association of HLA-DR genotypes and IL-1ra gene polymorphism with treatment failure of budesonide and disease patterns in Crohn's disease.

Objective Associations between HLA-DR genotypes and susceptibility to Crohns disease (CD) have been reported. However, it is not known whether certain HLA-DR genotypes or IL-1ra gene polymorphism are associated with responsiveness to treatment or different clinical patterns of disease. Design/setting In a large, randomized, controlled multicentre trial, 318 patients with CD were treated with daily doses of 6, 9 or 18 mg budesonide. Patients were stratified into two groups: patients without steroid pretreatment and with active CD (CDAI > 150) and patients with conventional steroid pretreatment of ⩽ 30 mg prednisolone per day, which was replaced by oral budesonide within 3 weeks. Main outcome measures The HLA-DRB1 genotypes 1–16 and the IL-1ra gene polymorphism were examined for an association with budesonide treatment failure. Results Only HLA-DR 8 was associated with treatment failure of budesonide. HLA-DR 8 is not very common. Only 17/243 patients who could be evaluated expressed this genotype, and 13 of these 17 patients did not respond to budesonide (P < 0.00067). Neither the other HLA-DR genotypes nor the IL-1ra gene polymorphism had an influence on treatment outcome of budesonide therapy. No significant association of fistulas, perianal disease, need for bowel resections, and disease localization with certain HLA-DRB1 genotypes or the IL-1ra gene polymorphism were found. Conclusions This is the first description of an association of a certain HLA-DR genotype (HLA-DR 8) with treatment failure in inflammatory bowel disease (IBD).

Eine neue Parazentesenadel für Aszites und Pleuraerguss im Vergleich mit der Venenverweilkanüle Eine prospektive, randomisierte Studie

ZusammenfassungHintergrund: Die diagnostische und therapeutische Punktion von Aszites oder Pleuraerguss gehört zur täglichen Routine auf internistischen und chirurgischen Klinikstationen. Die Parazentese wird in Deutschland in der Regel mit Venenverweilkanülen durchgeführt. Die therapeutische Punktion von Aszites oder Pleuraerguss mittels Venenverweilkanüle ist jedoch oft genug eine unbefriedigende, zeitraubende und oft begrenzt erfolgreiche Intervention. Gründe hierfür liegen hauptsächlich im für die Durchführung von Parazentesen ungeeigneten Material und in der Konzeption der Venenverweilkanüle.nn Patienten und Methode: 45 Patienten mit Aszites oder Pleuraerguss wurden randomisiert entweder mit einer von uns speziell für die Parazentese entwickelten neuen Punktionsnadel oder mit einem Venenverweilkanüle punktiert. Der Gebrauch beider Nadeltypen wurde unter folgenden Gesichtspunkten verglichen: Notwendigkeit und Zahl der Lageänderungen der Nadel während der Punktion, Notwendigkeit der Repunktion und Abbruchursache, Flussgeschwindigkeit, subjektive Praktikabilität der Parazentese und Patientenakzeptanz.nn Ergebnisse: Die Parazentesenadel war der Venenverweilkanüle unter allen untersuchten Aspekten deutlich überlegen. Insbesondere gelang die komplette Entlastung von Aszites und Pleuraerguss mit der Parazentesenadel signifikant häufiger als mit der Venenverweilkanüle.nn Schlussfolgerung: Die Anwendung der Parazentesenadel ist der der Venenverweilkanüle objektiv überlegen. Sie kann helfen, zusätzliche, durch Repunktionen ausgelöste Komplikationen zu vermeiden und den Patientenkomfort zu erhöhen.AbstractBackground: Diagnostic or therapeutic paracentesis of ascites or pleural effusions is part of the daily routine on many hospitals wards and in outpatient clinics. In Germany, paracentesis is usually performed with angiocaths. However, the therapeutic large volume paracentesis of ascites and paracentesis of pleural effusions with angiocaths is often cumbersome and quite often paracentesis fails, forcing the physician to repuncture. This is mainly due to the fact that angiocaths are not designed for such interventions.nn Patient and Method: 45 patients with ascites or pleural effusions were treated with a new needle especially designed for paracentesis, or with an angiocath. The new paracentesis needle was compared with the angiocath needle under the following aspects: necessity and number of positional corrections of the needle, necessity of and reasons for repuncture, duration of puncture, flow capacity, subjective practicability of paracentesis and patient acceptance of the paracentesis needle.nn Results: The paracentesis needle was superior to the angiocath in all investigated respects. Significantly, the paracentesis needle had a much higher success rate in the complete drainage than had the angiocath.nn Conclusion: The paracentesis needle was objectively superior as compared to the angiocath. It might help to avoid additional complications due to repuncture and it will increase the patients comfort.

Complicated wireless capsule enteroscopy in a patient with Crohn's disease.

An 18-year-old male experienced a first episode of Crohns disease with inflammation of the colon, stenosis of small intestine and a fistula of the anal sphincter. After resection of the fistula and 30 cm of proximal ileum the patient remained free of symptoms under medication with azathioprine and mesalazine for 6 years. Then, blood in the stool occurred. Diagnostic work-up - gastroscopy and colonoscopy 2004 and 2006, magnetic resonance enteroclysis 2004 and 2006 and wireless capsule enteroscopy 2006 - revealed slight inflammation in the ileum but no bleeding source. The bleeding ceased, but after 2 uneventful years abdominal cramps appeared and diagnostic work-up (magnetic resonance enteroclysis, radiograph) located the capsule still in the terminal ileum proximal to an inflamed stenosis. Corticosteroids were subscribed for 4 weeks, but the capsule stayed in place. Surgery was discussed, but denied by the patient. Finally, the capsule could be taken out by double balloon enteroscopy.

Palpatorische und sonographische Detektion von Lymphknotenmetastasen bei lokal fortgeschrittenem malignen Melanom

Zusammenfassung.Hintergrund und Ziel:Die Detektion von Metastasen beim malignen Melanom istntherapeutisch relevant. Ziel der Arbeit ist der Vergleich dernPalpation mit der Sonographie zur Lymphknotenbeurteilung beimnlokal fortgeschrittenen malignen Melanom.Patienten und Methodik:83 Patienten mit malignem Melanom (Clark-Level IV oder V)nwurden sowohl palpatorisch als auch sonographisch auf suspektenLymphknoten untersucht. Bei positiven Lymphknoten wurde dasnUntersuchungsergebnis mit dem histopathologischen Befund odernaber dem Befund bei der nächsten Visite verglichen.Ergebnisse:Lymphknotenmetastasen wurden histopathologisch bei 14nPatienten nach der ersten Studienuntersuchung, bei drei weiterennPatienten nach der Kontrolluntersuchung nachgewiesen.nSensitivität, Spezifität, positiver und negativer Vorhersagewertnfür den Nachweis von Filiae oder von größenprogredienten undnverdächtigen Lymphknoten bei der nächsten Untersuchung lagen fürndie Palpation bei 65%, 81%, 48% bzw. 89%, für dienLymphknotensonographie bei 100%, 66%, 45% bzw. 100%.Schlussfolgerung:Die Lymphknotensonographie sollte integraler Bestandteilnbei der Detektion von Lymphknotenmetastasen beim lokalnfortgeschrittenen malignen Melanom sein.Abstract.Background and Purpose:Early detection of metastases of malignant melanoma hasntherapeutic implications. The aim of this study was to evaluatenpalpation and ultrasound examination in the diagnostics of lymphnnode metastases in locally advanced melanoma.Patients and Methods:83 patients suffering from melanoma (Clark level IV or V)nwere examined for lymph node metastases by palpation andnsonography. Findings were compared to histopathologic resultsnafter lymph node extirpation if available or the findings at thennext follow-up visit.Results:Lymph node metastases were confirmed histopathologicallynin 14 patients at the first study visit, in three others at thencontrol visit. Sensitivity, specificity, positive and negativenpredictive values of palpation for the detection of metastasesnor suspicious nodes with increasing volume at follow-up in thisnpopulation were 65%, 81%, 48%, and 89%, and of ultrasound 100%,n66%, 45%, and 100%, respectively.Conclusion:Sonography of lymph nodes should be included as a standardnprocedure in the detection of metastases of locally advancednmalignant melanoma.

[Liver histology in hepatitis C: correlation with different biochemical and virological parameters]

ZusammenfassungHintergrund und Ziel: Die Indikation zur Therapie der HCV-RNA-positiven chronischen Hepatitis C ergibt sich nach den derzeitigen deutschen Leitlinien nicht aus der Histologie, sondern durch das Vorliegen erhöhter Transaminasen. Die Indikation zur Gewinnung einer Leberhistologie vor Therapiebeginn wird kontrovers diskutiert. Ziel dieser Studie war die Untersuchung der Korrelation verschiedener biochemischer und virologischer Parameter mit dem histologischen Entzündungs- und Fibrosegrad bei chronischer Hepatitis C.nn Patienten und Methodik: In einer retrospektiven Untersuchung wurden die Daten von 126 Patienten analysiert, bei denen zwischen Januar 1994 und März 1998 bei chronischer Hepatitis C eine Leberpunktion durchgeführt worden war. Die Histologien wurden einheitlich von einem Pathologen nach einem numerischen Entzündungs- und Fibrosegrad analysiert. Entzündungs- und Fibrosegrade wurden korreliert mit biochemischen und virologischen Parametern.nn Ergebnisse: Der Entzündungsgrad korrelierte mit der Höhe der GPT (r = 0,33; p < 0,001) nicht aber das Fibrosestadiulm (r = 0,15). Weder Entzündungs- noch Fibrosegrad zeigten eine signifikante Korrelation mit dem HCV-Genotype (n = 110) oder der HCV-RNA-Kopienzahl im Serum (n = 57). Entzündungs- und Fibrosegrad waren hochsignifikant miteinander assoziiert (p < 0,0001).nn Schlußfolgerung: Transaminasen spiegeln als “Surrogatmarker” in etwa die histologische Entzündungsaktivität wider, erlauben jedoch keine Rückschlüsse auf das Fibrosestadium. Bei einem Teil der Patienten können jedoch durchaus eine hohe Entzündungsaktivität bei niedrigen Transaminasen oder hohe Transaminasen bei niedriger histologischer Entzündungsaktivität vorliegen. Virologische Parameter wie HCV-Genotyp oder HCV-RNA-Kopienzahl im Serum lassen keine Rückschlüsse auf den Entzündungsgrad oder das Fibrosestadium zu. Der Verzicht auf eine Leberhistologie vor Einleitung der Therapie einer Hepatitis C und die ausschließliche Bestimmung von “Surrogatmarkern” können zu einer deutlichen Fehleinschätzung der tatsächlichen Entzündungs- und Umbauvorgänge in der Leber führen.AbstractBackground and Aim: According to the German consensus statement, the indication for treatment of HCV-RNA-positive chronic hepatitis C is not derived from histopathology but from elevated aminotransferases. The indication for liver biopsy has been discussed controversely. This study aimed at investigating the correlation between different biochemical and virological parameters and histological scores of inflammation and fibrosis in chronic hepatitis C.nn Patients and Methods: In a retrospective study, data of 126 patients with chronic hepatitis C who had undergone liver biopsy between January 1994 and March 1998 were analyzed. Histology was interpreted according to a defined numerical score of inflammation and fibrosis by a single pathologist. Scores of fibrosis and inflammation were correlated with biochemical and virological parameters.nn Results: Inflammatory grading showed a moderate but significant correlation with ALT (r = 0,33, p < 0.001), whereas staging of fibrosis did not correlate with ALT (r = 0.15). There was no association between grading or staging and HCV genotype (n = 110) or serum viral load (n = 57). Grading and staging showed a significant association with each other (p < 0.0001).nn Conclusion: Aminotransferases as “surrogate markers” reflect more or less the histological inflammatory activity but do not allow any estimation of the extent of fibrosis. Some patients may have a high inflammatory activity with low aminotransferases or high aminotransferases with low inflammatory activity. Virological parameters such as HCV genotype or viral load do not allow an estimation of histological findings. If prior to treatment of chronic hepatitis C liver biopsy is omitted and the decision for treatment depends solely on the measurement of surrogate markers, considerable misjudgment of the actual status of liver inflammation or fibrosis may result.