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Background Biliary obstruction and cholangitis are common problems in gastroenterology. Infections of the biliary tract with Candida and other fungal species have increasingly been seen in the last few years. Objective To investigate the prevalence, associations, and trends of biliary-tract candidiasis. Design A prospective, observational, diagnostic study. Setting University Hospital, Muenster, Germany. Patients Consecutive patients undergoing ERCP for various indications. Results In 54 of 123 patients, we found Candida species in bile samples (44%). In only 7 patients, candidiasis was suspected on endoscopy before mycologic proof. Only 4 of these 7 patients were correctly diagnosed with biliary candidiasis by simple morphologic aspects. The fungus was mainly differentiated as Candida albicans or Candida glabrata and rarely as Candida parapsilosis , Candida tropicalis, or other subspecies. Immunosuppression for various reasons was significantly associated with bile-duct candidiasis ( P P = .0824) or prior ERCP ( P = .1152). Biliary candidiasis was neither associated with positive fungal cultures of buccal smears ( P = .0722) nor with positive findings in stool samples ( P = .0860). Limitations Highly selected patient population. Buccal smears and stool samples were not obtained from all patients. Contamination artifacts cannot totally be excluded with the ERCP procedure. Conclusions Candida species very frequently can be detected in the bile. Positive fungal cultures of bile samples are not just contamination artifacts. This has to be taken into account when designing an anti-infectious treatment for recurrent cholangitis or even more cholangiosepsis. Especially in immunosuppressed patients or recipients of long-term antibiotic therapy, physicians should screen for biliary-tract candidiasis during endoscopic examination.

The combination of the recently introduced soft lithographic technique of inverted microcontact printing (i-muCP) and spin-coated films of polystyrene- block-poly( tert-butyl acrylate) (PS 690- b-P tBA 1210) as a reactive platform is shown to yield a versatile approach for the facile fabrication of topographically structured and chemically patterned biointerfaces with characteristic spacings and distances that cross many orders of magnitude. The shortcomings of conventional muCP in printing of small features with large spacings, due to the collapse of small or high aspect ratio stamp structures, are circumvented in i-muCP by printing reactants using a featureless elastomeric stamp onto a topographically structured reactive polymer film. Prior to molecular transfer, the substrate-supported PS 690- b-P tBA 1210 films were structured by imprint lithography resulting in lateral and vertical feature sizes between50 microm-150 nm and1.0 microm-18 nm, respectively. Time-of-flight secondary ion mass spectrometry (ToF-SIMS) and water contact angle measurements provided evidence for the absence of surface chemical transformations during the imprinting step. Following the previously established hydrolysis and activation protocol with trifluoroacetic acid and N-hydroxysuccinimide, amino end-functionalized poly(ethylene glycol) (PEG-NH 2), as well as bovine serum albumin and fibronectin as model proteins, were successfully transferred by i-muCP and coupled covalently. As shown, i-muCP yields increased PEG coverages and thus improved performance in suppressing nonspecific adsorption of proteins by exploiting the high local concentrations in the micro- and nanocontacts during molecular transfer. The i-muCP strategy provides access to versatile biointerface platforms patterned across the length scales, as shown for guided cancer cell adhesion, which opens the pathway for systematic cell-surface interaction studies.

To determine the impact of a broad spectrum of different polymorphisms within the interleukin-10 (IL-10) promoter gene on disease susceptibility to primary Sjogren's syndrome (pSS), clinical manifestations, and autoantibody production.We genotyped 111 unrelated German Caucasian patients with pSS and 145 healthy controls for the single nucleotide polymorphisms (SNPs) at positions -2849, -2776, -2769, -2763, -1349, -1082, -851, -819, -657, and -592 and for the microsatellites IL10.R and IL10.G. Allele and haplotype distributions were compared between patients and controls and between subgroups of patients with different clinical and laboratory findings.We found no significant differences in the allele or haplotype frequencies between pSS patients and healthy controls. After Bonferroni correction we found a significant association of the ACC haplotype (at the -1082, -819, and -592 loci) with immunoglobulin (Ig)A antibodies to anti-alpha-fodrin.Overall we found no associations of IL-10 promoter polymorphisms with the susceptibility to pSS in our cohort. The finding that the production of IgA anti-alpha-fodrin antibodies is associated with polymorphisms within the IL-10 promoter region suggests a genetic contribution to the generation of these antibodies.

Background: The understanding of genetic risk factors for chronic pancreatitis (CP) increased in the last decade with the discovery of mutations in the cationic trypsinogen gene (PRSS1). The first mutation was detected at the R122 autocleavage site of the protein (R122H) and subsequently two other mutations in this region, R122C and V123M, were described that resulted in a similar phenotype of hereditary pancreatitis. This study reports a novel A121T mutation within this region and characterizes the resulting molecular properties at the autocleavage site. Methods: Blood samples of a PRSS1 A121T carrier family were analyzed for PRSS1 mutations using melting point curve analysis, restriction endonucleases and DNA sequencing. Conformation dependent properties of the mutated sequence were analyzed by molecular modelling. The autodegradation kinetic of the mutated trypsin sequence was measured by a novel fluorescence resonance energy transfer (FRET) assay using designed 11 amino acid peptides from PRSS1 aa 118 - aa 127 containing the trypsin cleavage site at aa 122 coupled to a Dabcyl/EDANS FRET system. The kinetic of tryptic peptide cleavage was measured in a fluorescence ELISA reader. Results: DNA sequencing revealed a novel G to A transition at position 133279 of the published genomic sequence (#U66061 GenBank). The mutation results in an amino acid substitution of Alanine by Threonine at position 121 (A121T) of the cationic trypsinogen. Four additional mutation carriers could be identified among the relatives while only the first patient developed chronic pancreatitis. Molecular modelling of PRSS1 A121T revealed a change in the bond pattern between the R122 region and the calcium binding loop, whereas FRET assays showed an increased trypsin cleavage rate with a reaction kinetic elevated by more than 80 %. Conclusion: The novel PRSS1 A121T mutation highlights the surface exposed region PRSS1 A121-R122-V123 as a hotspot for hereditary pancreatitis associated trypsinogen mutations. Molecular modelling and FRET assays provide evidence for a A121T mutation dependent increase in susceptibility to trypsin digestion at the R122 cleavage site suggesting an enhanced autodegradation and a loss-of-function at the autocleavage site.

Diagnostic and therapeutic interventions in the biliary and pancreatic system in the previously operated patient by conventional endoscopic retrograde cholangiopancreaticography (ERCP) are difficult and, depending on the surgical procedure, in many cases unsuccessful. We describe our experience of ERCP performed with a double balloon enteroscope (DBE) as an alternative examination technique for these patients.In a retrospective analysis of all DBE procedures at our department between November 2004 and June 2007, 11 patients were identified with various anatomic variations in whom ERCP was performed using a DBE.In 72% of the patients, previous conventional ERCP examinations failed (8/11). In these patients, DBE-ERCP was successful in 63%. The overall success rate of DBE-ERCP in all patients was 64% (7/11 patients). In those patients, interventions such as papillotomy, calculus extractions, as well as stent placement could be performed even though tools for DBE-ERCP are still very limited. Despite most of the DBE-ERCPs having included therapeutic interventions, no major complications occurred in our case series and minor side effects were restricted to meteorism and mild to moderate abdominal pain.DBE-ERCP is an alternative method for diagnostic as well as therapeutic interventions in the biliary as well pancreatic system in the operated patient. However, it should be limited to selected patients, e.g., with contraindications for PTC, as it is a time-consuming as well as a cost-intensive procedure.

BACKGROUND Undifferentiated embryonal sarcoma of the liver (UESL), a rare tumor that predominantly affects children, generally has been considered an aggressive neoplasm with an unfavorable prognosis. More recent reports have indicated that modern multimodal treatment and supportive care improve the survival of children with UESL. Data regarding the treatment and survival of adults have not been reviewed comprehensively, and only a few adult patients with UESL have been reported in the literature. METHODS The authors analyzed demographics, treatment, and actuarial survival of all reported cases of UESL in patients aged ≥15 years (n = 67 patients). In addition, 1 case is presented of a patient with UESL who was treated successfully at the authors' institution. RESULTS The median survival of all patients with UESL who were analyzed was 29 months. Patients who underwent complete tumor resection followed by adjuvant chemotherapy survived over a median follow-up of 28.5 months and had significantly better survival compared with patients who underwent surgical treatment alone. Patients who underwent an incomplete tumor resection had a tendency toward poorer outcomes. CONCLUSIONS To the authors' knowledge, this is the first report to demonstrate a significant effect on survival for adjuvant chemotherapy after complete surgical resection of UESL in adults. The role of neoadjuvant chemotherapy was not evaluated in this study. In the case study presented herein, combined therapy with surgery and chemotherapy led to a complete, sustained remission that has lasted for >6 years to date. Cancer 2008. © 2008 American Cancer Society.

This prospective single-centre phase II trial assessed the diagnostic impact of 18F-FDG PET–CT in the evaluation of solid pancreatic lesions (∅ ≥10 mm) compared to endosonography (EUS), endoscopic retrograde cholangio-pancreatography (ERCP) with intraductal ultrasound (IDUS), abdominal ultrasound (US) and histopathological reference. Forty-six patients (32 men/14 women, ∅ 61.7 years) with suspected pancreatic neoplasms underwent PET–CT with contrast-enhanced biphasic multi-detector CT of the upper abdomen followed by a diagnostic work-up with EUS, ERCP with IDUS and US within 3 weeks. PET–CT data sets were analysed by two expert readers in a consensus reading. Histology from surgery, biopsy/fine-needle aspiration and/or clinical follow-up ≥12 months served as standard of reference. Twenty-seven pancreatic malignancies were histopathologically proven; 19 patients had benign diseases: 36/46 lesions (78%) were detected in the head of the pancreas, 7/46 and 3/46 in the body and tail region, respectively. Sensitivity and specificity of PET–CT were 89% and 74%, respectively; positive predictive value (PPV) and negative predictive value (NPV) were 83% and 82%, respectively. Sensitivity (81–89%), specificity (74–88%), PPV (83–90%) and NPV (77–82%) achieved by EUS, ERCP and US were not significantly different. PET analysis revealed significantly higher maximum mean standardised uptake values (SUVmax 6.5 ± 4.6) in patients with pancreatic malignancy (benign lesions: SUVmax 4.2 ± 1.5; p

Citrobacter rodentium induces an acute, self-limited colitis in mice which is histologically associated with crypt hyperplasia. The infection serves as a model for human infectious colitis induced by enteropathogenic Escherichia coli. We investigated if Balb/c mice, which had spontaneously cleared C. rodentium infection, were protected against re-infection and if resistance against intestinal infection can be systemically transferred using spleen cells. The course of infection was monitored by faecal excretion. Spleen cells, splenic CD3+ and CD4+ cells were transferred from resistant mice to non-infected recipients prior to infection. Cytokine secretion, serum and faecal antibody titres and histological disease severity were assessed. Balb/c mice were resistant against re-infection. The course of infection was shorter in mice receiving primed spleen cells, CD3+ and CD4+ cells. Transfer of CD4+ T cells from resistant mice induced gamma-interferon, interleukin (IL)-2 and IL-17 secretion and suppressed IL-10 secretion. Anti-Citrobacter serum IgG1 and IgG2a enzyme-linked immunosorbent assay OD levels were increased. Faecal IgA secretion was increased while serum IgA was suppressed in recipients of CD4+ cells. Large bowel histology showed protection from colitis in recipients of primed cells as indicated by normal colonic epithelium. In Balb/c mice, C. rodentium infection is followed by resistance, which can be transferred by CD4+ cells. Transfer of protection is associated with IL-17 secretion, enhanced serum IgG and faecal IgA secretion. This is the first study to demonstrate the mechanisms by which systemic resistance from previously C. rodentium-infected mice can be transferred to non-infected animals.

One hundred symptomatic patients with radiolucent gallbladder stones were treated with a new piezoelectric lithotripter and oral chemolitholytic agents. Stone disintegration was achieved in 99 of these patients (99%) with a mean (+/- SD) maximum fragment size of 5.1 +/- 4.1 mm. Significant differences were found when the mean (+/- SD) fragment sizes of single stones less than or equal to 20 mm (4.2 +/- 2.5 mm) were compared with those of single stones greater than 20 mm (5.8 +/- 3.4 mm; P less than 0.05) and multiple stones (6.2 +/- 3.8 mm; P less than 0.05), respectively. None of the patients required anesthesia, analgesics, or sedatives before or during the treatment. The stone-free rates for all patients followed up for up to 4-12 months (mean +/- SD, 10.7 +/- 2.9 months) were 18% (1 month), 25% (2 months), 38% (4 months), 52% (8 months), and 67% (12 months). Partly significant differences were obtained in stone-free rates for single stones (less than or equal to 20 mm) compared with larger stones (greater than 20 mm) and multiple stones (P less than 0.05), respectively. Serious adverse reactions (i.e., cholestasis and pancreatitis) were observed in only 3 patients (3%). These conditions were induced by fragment impaction in the common bile duct. In 2 of these patients, endoscopic retrograde cholangiopancreatography with endoscopic sphincterotomy was required. It is concluded that piezoelectrically generated shock waves are suitable for the effective and safe disintegration of gallbladder stones in humans. The anesthesia-free and analgesia-free shock-wave application opens up the possibility to perform biliary lithotripsy as an outpatient procedure. The stone-free rate achieved in combination with oral bile acids is most promising for single stones (less than or equal to 20 mm).

A randomised controlled multicentre trial was performed in 160 patients with gastric ulcer, proved by endoscopy and biopsy, to compare ulcer healing with sucralfate and ranitidine (double blind double dummy design) and to assess the effect of maintenance treatment with sucralfate on ulcer recurrence (double blind placebo controlled design). The healing rates were similar with 4 g sucralfate suspension per day and 300 mg ranitidine per day (82% and 88% after 12 weeks, respectively). Of the 109 patients with healed ulcers, 92 were entered into the maintenance trial and treated with sucralfate tablets (2 g per day) or placebo tablets. Maintenance treatment with sucralfate delayed symptoms of gastric ulcer recurrence. Lifetable analysis showed significant differences between sucralfate and placebo, both after six months (p = 0.018) and after 12 months (p = 0.044). The rates of symptom recurrences were 13% and 34% after six months and 34% and 55% after 12 months for sucralfate and placebo, respectively. The rate of asymptomatic recurrences after 12 months was similar in the two groups (9% and 10%, respectively). The recurrence rate was higher in patients who had never taken non-steroidal anti-inflammatory drugs than in those who had but had stopped on admission to the study. It was also higher in patients with recurrent ulcer and in those with scarring deformation and narrowing of the pylorus. Maintenance treatment with sucralfate slowed the appearance of symptom recurrences of gastric ulcer.

Einleitung: Der Chemokinrezeptor CCR6 ist essentiell fur die Entwicklung des organisierten Immunsystems des Darmes (gut associated lymphatic tissue, GALT), seine Deletion fuhrt zu einer funktionellen Beeintrachtigung der Peyer'schen Plaque bzw. isolierter lymphatischer Follikel mit deutlich reduzierter Anzahl von M-Zellen im Follikel-assoziierten Epithel. Da das organisierte GALT an der Entwicklung chronisch-entzundlicher Darmerkrankung beteiligt ist, soll der Einfluss der CCR6-Deletion auf die Entwicklung einer chronischen Kolitis untersucht werden. Methodik: Zur Beurteilung der Bedeutung von CCR6 fur die Entwicklung einer chronischen Kolitis wurden IL-10-/- CCR6-/- doppeldefiziente Mausen generiert und mit IL-10 defizienten Mausen verglichen. Die Erkrankungsaktivitat wurde anhand des Gewichtsverlaufs, der Entwicklung eines Prolaps bzw. der spontanen und Piroxicam-induzierten Letalitat bestimmt. Nach Entwicklung einer manifesten Kolitis erfolgte eine histologische Beurteilung des gesamten Kolons. Die Zytokinexpression (Interferon-γ, TNF) wurde in CD3-stimulierten Lymphozyten aus der Milz bzw. den mesenterialen Lymphknoten untersucht. Ergebnisse: IL-10 defiziente C57BL/6-Mause entwickeln eine spontan nicht-letale chronische Kolitis im Verlauf von 10 Wochen, wahrend CCR6-defziente Mause keine Zeichen der Kolitis aufweisen. IL-10-/- CCR6-/- doppeldefiziente Mause zeigen dagegen eine sich deutlich fruher entwickelnde Kolitis mit beeintrachtigter Gewichtszunahme ab der 4 Woche, fruhem Analprolaps und signifikanter spontaner (50% vs. 5%) bzw. Piroxicam induzierter Letalitat (100% vs. 15%). Histologisch zeigt sich eine deutlich vermehrte Epithelzellhyperplasie bei vergleichsweise reduziertem lymphatischen Infiltrat. Dagegen kann nach CD3-Stimulation von Lymphozyten aus mesenterialen Lymphkonten eine geringere Sekretion TH-1 spezifischer Zytokine nachgewiesen werden. Diskussion: Der Chemokinrezeptor CCR6 ist fur die Entwicklung intestinaler Immunantworten essentiell. Seine Deletion fuhrt zu einer Akzentuierung der klinischen Aktivitat bzw. Letalitat einer chronischen Kolitis, die moglicherweise Folge einer Inhibition des organisierten GALT ist.

The objective of this study was to analyse levels of the proinflammatory cytokine macrophage migration inhibitory factor (MIF) in patients with primary Sjögren's syndrome (pSS) and to examine associations of MIF with clinical, serological and immunological variables. MIF was determined by ELISA in the sera of 76 patients with pSS. Further relevant cytokines (IL-1, IL-6, IL-10, IFN-gamma and TNF-alpha) secreted by peripheral blood mononuclear cells (PBMC) were determined by ELISPOT assay. Lymphocytes and monocytes were examined flow-cytometrically for the expression of activation markers. Results were correlated with clinical and laboratory findings as well as with the HLA-DR genotype. Healthy age- and sex-matched volunteers served as controls. We found that MIF was increased in patients with pSS compared with healthy controls (p0.01). In particular, increased levels of MIF were associated with hypergammaglobulinemia. Further, we found a negative correlation of MIF levels with the number of IL-10-secreting PBMC in pSS patients (r = -0.389, p0.01). Our data indicate that MIF might participate in the pathogenesis of primary Sjögren's syndrome. MIF may contribute to B-cell hyperactivity indicated by hypergammaglobulinemia. The inverse relationship of IL-10 and MIF suggests that IL-10 works as an antagonist of MIF in pSS.

Despite tremendous effort and progress in the diagnostics of pancreatic cancer with respect to imaging techniques and molecular genetics, only very few patients can be cured by surgery leading to a 5-year survival rate of only 3%. Especially the lack of chemotherapeutical options in this entity requires a better understanding of the molecular mechanisms leading to pancreatic carcinoma growth and progression in order to develop novel treatment regimens. To identify signaling pathways that are critical for this tumor entity, we compared six well-established pancreatic cancer cell lines (Capan-1, Capan-2, HUP-T3, HUP-T4, KCL-MOH, PaTu-8903) with colon cancer cell lines and tumor cell lines of non-epithelial origin by expression profiling. For this purpose we employed Human Genome Focus Arrays representing about 8500 well annotated human genes. We identified 353 genes with significantly high expression in the group of pancreatic carcinomas. Based on Gene Ontology annotations these genes are especially involved in Rho protein signal transduction, proteasome activator activity, cell motility, apoptotic program, and cell-cell adhesion processes indicating these pathways to be interesting candidates for the design of targeted therapies. Most pancreatic carcinomas are characterized by mutations in the TP53 and the KRAS genes and the absence of microsatellite instability, which could also be confirmed for our panel of pancreatic carcinoma cell lines. Looking for individual differences within this group that may be responsible for more or less aggressive behavior, we identified genomic amplifications at the 8q22.1 and the 8q24.22 loci to be associated with enhanced gene transcription. Because we have previously shown that gains of genomic material from the long arm of chromosome 8 have an adverse effect on the outcome of pancreatic carcinoma patients, we conclude that functional analysis of amplified genes at 8q22 and/or 8q24 may lead to an improved understanding of pancreatic carcinoma progression.

Highly localized, selective deprotection chemistry and efficient grafting reactions in microcontacts between elastomeric stamps and reactive polystyrene-block-poly(tert-butyl acrylate) (PS690-b-PtBA1210) diblock copolymer films are developed. The procedure yields well-defined protein-poly(ethylene glycol) (PEG) nanopatterns (see figure), which may find applications in, for example, cancer-cell-surface interaction studies.

The inflammatory bowel diseases (IBD) are comprised of two major phenotypes, Crohn's disease (CD) and ulcerative colitis (UC). Research over the last couple of years has led to great advances in understanding the inflammatory bowel diseases and their underlying pathophysiologic mechanisms. From the current understanding, it is likely that chronic inflammation in IBD is due to aggressive cellular immune responses to a subset of luminal bacteria. Susceptibility to disease is thereby determined by genes encoding immune responses which are triggered by environmental stimuli. Based on extensive research over the last decade, there are several new and novel pathways and specific targets on which to focus new therapeutics. The following review summarizes the current view on the four basic tenets of the pathophysiological basis of IBD and its implications for therapies of IBD: genetics, immune dysregulation, barrier dysfunction and the role of the microbial flora.

Hallmarks of the adaptive immune system are antigen-specific cellular and humoral immune responses. Secondary lymphoid organs serve as sites of contact between antigen-presenting cells (APCs) and immune effector T and B lymphocytes. The gut-associated lymphatic system (GALT) as the intestinal branch of the immune system provides different mechanisms to protect organisms against pathogens. Simultaneously, immune activation secondary to genetic factors and/or environmental signals can induce detrimental autoimmunity. The effector pathways in host defense and autoimmunity use similar cytokines and chemokines. Unlike few other cytokines, lymphotoxin (LT) alpha/beta regulates the development of intestinal lymphoid organs, including Peyer's patches (PPs) and mesenteric lymph nodes (MLNs). In addition, intestinal inflammation is suppressed by inhibition of LTbeta signalling, an observation which has initiated clinical studies using this treatment principle. Conversely, the course of Citrobacter rodentium-induced infectious colitis is more severe in mice with impaired LTbeta-receptor-mediated signalling. This report provides an overview on the role of the different organs of the GALT in intestinal inflammation. Moreover, it describes the role of the LTbeta-receptor-mediated signalling in intestinal inflammation as encountered in autoimmune and infectious pathology. The contribution of LT to the delicate balance of immune effector functions in host defense and autoimmunity is discussed.

Background and aims: α-Melanocyte stimulating hormone (αMSH) is known to exert anti-inflammatory effects, for example in murine DSS (dextran sodium sulphate induced) colitis. The anti-inflammatory functions of αMSH are mediated by the melanocortin1-receptor (MC1R) in an autoregulatory loop. The aim of this study was therefore to determine whether a breakdown of the αMSH–MC1R pathway leads to worsening of disease. Methods: Experimental colitis was induced in mice with a frameshift mutation in the MC1R gene (MC1Re/e), C57BL/6 wild type mice, and MC1Re/e-C57BL/6 bone marrow chimeras. The course of inflammation was monitored by weight loss, histological changes in the colon, and myeloperoxidase activity. In addition, MC1R expression was analysed in intestinal epithelial cells. Results: While the colon of untreated MC1Re/e appeared normal, the course of DSS-colitis in MC1Re/e mice was dramatically aggravated, with a significantly higher weight loss and marked histological changes compared to C57BL/6WT. The inflammation eventually led to death in all MC1Re/e, while all C57BL/6WT survived. Similar observations were detected in a transmissible murine colitis model induced by Citrobacter rodentium . Infected MC1Re/e showed delayed clearance of infection. To determine whether missing haematopoietic cell expressed MC1R was responsible, DSS colitis was induced in MC1Re/e-C57BL/6 bone marrow chimeras. MC1Re/e mice receiving MC1R+ bone marrow showed a similar course of inflammation to non-transplanted MC1Re/e. Likewise, transplantation of MC1R bone marrow into C57BL/6WT mice did not lead to any worsening of disease. Conclusions: This is the first study to show a functional role of MC1R in intestinal inflammation. The data suggest a pivotal role of non-haematopoietic cell expressed MC1R in the host’s response to pathogenic stimuli.

Einleitung: Mikrovaskulare Endothelzellen stellen eine sehr heterogene Zellpopulation in Abhangigkeit von ihrem Ursprungsgewebe dar. Die Mikrovaskulatur des Magens spielt eine wichtige Rolle sowohl bei Verdauungsprozessen als auch im Rahmen von Entzundungsreaktionen und in der Tumorentstehung. Methoden zur Isolierung und Kultivierung spezifischer Endothelzellen sind bereits fur andere Organsysteme etabliert worden. Im Folgenden beschreiben wir eine neue Methode zur Isolierung von magenspezifischen mikrovaskularen Endothelzellen. Methoden und Material: HuGE wurden aus gesundem Randgewebe von Tumorgastrektomiepraparaten gewonnen. Nach mechanischer und enzymatischer Aufbereitung mit Kollagenase wurden kleine, mikroskopisch identifizierte Endothelzellcluster auf kollagen-beschichteten Kulturschalen kultiviert. Kontaminierende Zellen wie Fibroblasten und glatte Muskelzellen wurden mechanisch entfernt. Ausreichend grose Endothelzellkolonien wurden auf Kulturplatten bzw. Gewebekulturflaschen transferriert. Die Reinheitskontrolle der Zellen erfolgte flowzytometrisch. Um den endothelialen Ursprung der Zellen zu beweisen, wurde von-Willebrand-factor (vWF) immunhistochemisch und die ICAM-1-Expression nach proinflammatorischer Stimulation flowzytometrisch bestimmt. Ergebnisse: Die nach dieser Methode isolierten HuGE wuchsen in Monolayern und zeigten lichtmikroskopisch ein typisches “Kopfsteinpflaster“-Muster. Flowzytometrisch konnte eine Reinheit von mehr als 95% nachgewiesen werden. Immunhistochemisch zeigte sich eine zytosolische Expression von vWF. Nach proinflammatorischer Stimulation mit TNF-α resultierte eine Hochregulation von ICAM-1. Die entstandenen HuGE-Monolayer-Kulturen waren mikroskopisch frei von Kontamination durch Fibroblasten oder glatte Muskelzellen. Eine Kultivierung gelang bis zur Passage 15. Diskussion: In dieser Arbeit wurde eine effektive Methode zur Isolation und Kultivierung von humanen Magenendothelzellen etabliert. Diese Methode ist kostensparend im Vergleich zu anderen zuvor beschriebenen Methoden auf der Basis von magnetischen Beads und ermoglicht die Herstellung sehr reiner primarer Magenendothelzellkulturen, welche zur Beantwortung einer Vielzahl biochemischer und zellbiologischer Fragestellungen eingesetzt werden konnen.

Angiogenesis is a crucial process involved in numerous physiological and pathophysiological settings. During the past three decades, the field of tumour associated angiogenesis has been the focus of a plethora of basic research projects and clinical studies. Tumour associated neovessels satisfying the increased demand of oxygen and nutrients in malignant tumours are now emerging as specific targets for novel antineoplastic agents. This article discusses the present data on antiangiogenic treatment of human gastrointestinal malignancies, including gastric, pancreatic, and colorectal adenocarcinoma, and outlines potential future perspectives, such as development of surrogate markers of angiogenesis.

Objectives. To analyse the prevalence of � -fodrin antibodies in patients with primary (pSS) and secondary Sjogren's syndrome (sSS) and the relation to clinical, serological and immunological features. Methods. Serum IgA and IgG antibodies to the 120 kDa � -fodrin were determined by ELISA technique in 62 pSS patients and 28 sSS patients. Results were correlated with clinical symptoms and laboratory findings as well as with the HLA-DR genotype. Additionally, antibody concentrations were correlated with the numbers of peripheral blood mononuclear cells (PBMCs) secreting interleukin (IL)-6, IL-10, interferon-� (INF)-� , and tumour necrosis factor-� determined by ELISPOT analysis. Lymphocytes and monocytes were examined flow-cytometrically for the expression of activation markers. Healthy age- and sex-matched volunteers served as controls. Results. The sensitivity of IgA and IgG � -fodrin antibodies was 35 and 31%, respectively, in pSS patients. In sSS patients, the sensitivity was 29 and 21%, respectively. In pSS patients with IgG antibodies, recurrent parotid swelling was significantly more prevalent. Also the number of INF-� secreting PBMCs and the percentage of CD4/CD71þ lymphocytes as well as CD14/ HLA-DRþ monocytes were significantly increased in this group compared with � -fodrin-negative patients or with controls. Interestingly, these patients also had a shorter disease duration. No association of � -fodrin antibodies with the HLA-DR genotype was found. Conclusion. Due to the low prevalence, serum antibodies to � -fodrin turned out to be of limited diagnostic value in our study. However, our data suggest that IgG antibodies to � -fodrin are indicative of clinical and immunological activity in pSS especially in patients with shorter disease duration and may thus serve as a marker of disease activity.

Background: Recent reports have shown an upregulation of macrophage migration inhibitory factor (MIF) during gastric ulcer development in a rat model and elevated counts of MIF-positive cells in biopsies from Helicobacter pylori-infected patients. H. pylori infection is a proven cofactor in humans causing gastritis and gastric ulcers. The aim of this study was to characterize MIF expression in human gastric epithelial cells in response to H. pylori. Methods: MIF mRNA and MIF protein expression was detected in human gastric epithelial cell lines after stimulation with proinflammatory cytokines or infection with H. pylori (cagA+/vacA+) using real-time reverse transcriptase–polymerase and enzyme-linked immunosorbent assay. Interleukin-8 secretion was measured as positive control. MIF mRNA and MIF protein expression was assessed in H. pylori-positive and -negative human gastric biopsy samples. Results: While interleukin-8 mRNA expression and interleukin-8 secretion were upregulated in gastric epithelial cells in vitro after H. pylori infection, no changes in MIF mRNA expression and MIF secretion could be detected. We found no significant differences in MIF expression in total RNA extracted from gastric biopsy tissue when comparing H. pylori-positive to control patients. Likewise, MIF protein expression in gastric epithelium was unaffected by H. pylori infection as compared to uninfected tissue. Conclusions: While an increased MIF expression and positive effects of MIF blockade in ulcer healing have been shown in a rodent model and elevated numbers of MIF-positive cells have been found in H. pylori-infected human tissue, we herein could not confirm any differences in human gastric epithelial MIF expression and secretion after H. pylori infection in vitro and in vivo.