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Biomedical subjects

Gerhard Rogler

Publications and source records attributed to Gerhard Rogler.

At least 37 records · Page 2Linked to original sources
PubMed2005

Association of antibodies to exocrine pancreas with subtypes of Crohn's disease.

OBJECTIVES: The Vienna classification of Crohn's disease (CD) subdivides patients according to their age at diagnosis, disease location, and disease behaviour. This study tested whether antibodies against exocrine pancreatic tissue (PAbs) are associated with Vienna classification subtypes of CD. METHODS: PAbs were examined by indirect immunofluorescence in sera of patients with CD (n = 208), or ulcerative colitis (n = 47), and in normal controls (n = 50). Presence of PAbs was compared to the Vienna classification, disease activity, and drug therapy. RESULTS: PAbs were present in 60 CD patients (28.8%), but in no patient with ulcerative colitis or in controls (P < 0.001). There was no significant association between PAbs and disease activity, or drug therapy. Complete classification into Vienna subtypes was possible in 168 patients. There was significant heterogeneity in PAb prevalence in these CD subtypes (P < 0.01). Variation was found in the behaviour category (B1, non-stricturing, non-penetrating CD: 14.0% PAb-positive; B2, stricturing CD: 41.7%; B3, penetrating CD: 31.5%; P = 0.02), but not age or location categories. PAbs were associated with long disease duration (P = 0.04). CONCLUSION: This study confirms that PAbs are highly specific for CD. The small differences in their prevalence in CD subtypes do not suggest that PAb detection is useful in discrimination of CD phenotypes.

Adult↗
PubMed2005

Crohn's disease patient characteristics in a tertiary referral center: comparison with patients from a population-based cohort.

OBJECTIVES: Data on the clinical presentation, effects of therapy and prognosis of patients with Crohn's disease are often based on patients from specialized referral centers. We assessed the extent of the selection based on the clinical and demographic characteristics. METHODS: All patients with Crohn's disease presenting to the University Hospital of Regensburg (Medical Department) were analyzed retrospectively with respect to demographic and disease specific characteristics. Only patients diagnosed <2 years before presentation were included in the main analysis. The original data from a population-based, prospectively assembled incidence cohort were available for comparison (EC-IBD, northern centers only, n=475). Age at diagnosis, disease location and behavior were categorized according to the Vienna classification. Differences were examined using chi-square tests. MAIN RESULTS: At the referral center, 394 patients were treated within a 5-year period. Of these, 116 patients fulfilled the inclusion criteria for the comparative analysis. Sixteen percent of the referral patients were diagnosed at age 40 or older, as compared with 32% in the population-based group (P<0.004). The distribution of disease location, sex, smoking behavior and positive family history was similar in both groups. Among the referral patients, more had fistulas (39% versus 20%, P<0.001). Also, more patients were receiving steroids (49% versus 27%) or other immunosuppressive therapy (12% versus 4%). The selection effects increase with duration of disease. CONCLUSIONS: Patients with late onset of disease, inflammatory only disease behavior and no need for immunosuppression are under-represented at a tertiary referral center.

Adolescent↗
PubMed2005

Dark lumen magnetic resonance enteroclysis in combination with MRI colonography for whole bowel assessment in patients with Crohn's disease: first clinical experience.

BACKGROUND: Magnetic resonance enteroclysis (MRE) is a recently introduced imaging technique that assesses the small bowel with similar sensitivity and specificity as the fluoroscopically performed conventional enteroclysis. Magnetic resonance imaging colonography (MRC) seems to be a promising technique for polyp assessment in the colon. In this feasibility study, we evaluated the combination of small bowel MRI with unprepared MRC as an integrative diagnostic approach of the whole bowel in patients with Crohn's disease. METHODS: Thirty patients with known Crohn's disease were prospectively examined. No particular colonic preparation was applied. Applying the dark lumen technique in all patients, MRE and MRC were performed within 1 session using an integrative examination protocol. T2-weighted and contrast-enhanced T1-weighted sequences were acquired. Inflammation assessment (grades 0 to 2) of the colon was compared with conventional colonoscopy in 29 patient and with surgery in 1 patient. The entire colon was graded fair to good distended in all patients. In 11 of 210 evaluated colonic segments, feces hindered an adequate intraluminal bowel assessment. Twenty-three of 30 patients had complete colonoscopy as the gold standard. In 7 patients, complete colonoscopy could not be performed because of an inflamed stenosis. RESULTS: Correct grading of colonic inflammation was performed with 55.1% sensitivity and 98.2% specificity in all segments. Considering only more extensive inflammation (grade 2), the sensitivity of MRC increased to 70.2% with a specificity of 99.2%. CONCLUSIONS: The combination of MRE and MRC could improve the diagnostic value of abdominal MRI evaluation in patients with Crohn's disease. However, MRC can not replace conventional colonoscopy in subtle inflammation assessment.

Adolescent↗
PubMed2005

The effect of chromoendoscopy on the diagnostic improvement of gastric ulcers by endoscopists with different levels of experience.

BACKGROUND: In recent years chromoendoscopy has become popular as a diagnostic enhancement tool in endoscopy. Using the macroscopic description of gastric ulcers, experienced endoscopists may be able to differentiate malignant and benign lesions. The aim of our study was to determine whether indigo carmine staining improves the ulcer differentiation by experienced and inexperienced endoscopists. METHODS: 50 patients were enrolled, 7 with malignant gastric ulcers and 43 with benign gastric ulcers. Gastroscopy was initially videotaped native, then on a second tape after staining with 0.2% indigo carmine. Later on biopsies were taken for histology. Subsequently the tapes were randomly evaluated by three experienced (>2000 gastroscopies; group A) and by three inexperienced (<100 gastroscopies; group B) investigators blinded from any personal data of the patients. The investigators had to classify the ulcers, using published criteria, native as well as stained. The results were compared within each group and with the histology. RESULTS: The endoscopic native diagnosis showed a sensitivity of 66.3%, a specificity of 86.3%, a positive predictive value of 48.1% and a negative predictive value of 94% for group A, respectively 66%, 62.5%, 22.7% and 92% for group B. After staining, the values of these parameters were reduced insignificantly to a sensitivity of 60.2%, a specificity of 78.5%, a positive predictive value of 36.1% and a negative predictive value of 92.8% for group A. Group B, on account of one investigator who demonstrated excellent skills, showed a significant better sensitivity (79.9%) and a slight improvement of the positive and negative predictive values to 25.7% respectively 94.8%, whereas the specificity very slightly decreased to 61.3%. The diagnostic accuracy before and after staining was 83.6%, respectively 76.5%, in group A and 63.2%, respectively 63.9% in group B. The correlation with the histology, determined by Cohen's kappa coefficient (median value), decreased from 0.46 for the native to 0.30 for the chromoendoscopic diagnosis in group A and remained unchanged (0.17) in group B. CONCLUSION: We concluded that chromoendoscopy does not improve the classification of gastric ulcers with respect to malignant or benign origin. The role of endoscopic experience could only be proved in the native macroscopic diagnosis of the investigators. After staining, with the exception of one investigator, experienced as well as inexperienced endoscopists lost their diagnostic accuracy.

Coloring Agents↗
PubMed2004

Bile salt-induced apoptosis in human colon cancer cell lines involves the mitochondrial transmembrane potential but not the CD95 (Fas/Apo-1) receptor.

BACKGROUND AND AIMS: Depending on their physico-chemical characteristics, bile acids can be potent inducers of apoptosis in colon cancer cells. This observation contrasts with bile acids being promoters of colorectal cancer carcinogenesis. Our recent observation of caspase activation in deoxycholate (DC)-treated colon cancer cell lines prompted us to analyze the mechanisms of bile acid-induced colon cancer cell death. METHODS: CD95 expression was correlated to DC-induced cell death in four colon cancer cell lines. Mitochondrial transmembrane potential (MTP) was determined in whole cells as well as in isolated mitochondria. RESULTS: On 2 of the 4 human colon cancer cell lines investigated, no CD95 was detected. These data were supported by a lack of CD95 mRNA in those cell lines that did not express CD95 on their surface. The apoptotic response to bile acids did not correlate with CD95 receptor expression on the respective cell lines. Therefore, we analyzed the MTP after the addition of toxic bile acids. MTP was destabilized early after the addition of deoxycholate to SW480 cells. These data were confirmed in isolated mitochondria, which showed strong swelling after the addition of DC. Accordingly, release of cytochrome-c from the mitochondrial intermembrane space into the cytosol, indicating dissipation of the MTP, and subsequent caspase-3 cleavage were detectable as early as 3 min after the addition of DC. CONCLUSION: In contrast to hepatocytes and hepatic carcinoma cell lines, DC induces apoptosis in colon cancer cell lines via a CD95 receptor-independent mechanism. Direct induction of the mitochondrial permeability transition by toxic bile acids is suggested as the apoptosis-inducing mechanism in colon cancer cells.

Apoptosis↗
PubMed2004

Both donor and recipient NOD2/CARD15 mutations associate with transplant-related mortality and GvHD following allogeneic stem cell transplantation.

Single nucleotide polymorphisms (SNPs) of the NOD2/CARD15 gene resulting in a diminished nuclear factor-kappaB (NF-kappaB) response to bacterial cell wall products have been associated with an increased incidence of Crohn disease. To assess a possible contribution of NOD2/CARD15 mutations to graft-versus-host disease (GvHD) and complications following allogeneic stem cell transplantation, we retrospectively typed DNA from donor/recipient pairs in 169 consecutive patients receiving transplants from related or unrelated donors. Mutated alleles were observed in 21% of patients and in 14% of donors. Cumulative incidence of 1-year, transplant-related mortality rose from 20% in donor/recipient pairs without mutated SNPs to 49% in pairs with recipient mutations (P =.03) and 59% in pairs with donor mutations (P <.005), and was highest in 12 pairs with mutated alleles in both donor and recipients (83%; P <.001). Similar associations were observed for severe overall and severe gastrointestinal GvHD. The impact of NOD2/CARD15 mutations was more prominent for HLA-identical sibling transplantations but was also observed in unrelated donor transplantation. Mutations proved to be independent risk factors for transplant-related mortality. Our findings indicate a major role of monocyte/macrophage dysfunction in the pathophysiology of GvHD and strongly suggest a future risk assessment or even donor selection through NOD2/CARD15 typing.

Adolescent↗
PubMed2004

[Extraintestinal manifestations of inflammatory bowel disease].

Extraintestinal manifestations of Crohn's disease and ulcerative colitis are found in > 50% of all patients. These extraintestinal manifestations sometimes impair the overall life quality much more than the bowel-related symptoms. Extraintestinal manifestations need to be distinguished from secondary diseases or complications of inflammatory bowel diseases, as they require a different and specific therapy. Complications of the intestinal disease, such as vitamin deficiency or osteoporosis, can be treated specifically by substitution of vitamin D, calcium, or other vitamins. However, extraintestinal manifestations of Crohn's disease and ulcerative colitis, such as primary sclerosing cholangitis, arthritis or granulomatous inflammation of the skin, lung, or liver, are much more difficult to treat sufficiently. Almost every organ can be a localization of extraintestinal symptoms of inflammatory bowel diseases. It is important to acquire knowledge on these extraintestinal manifestations of Crohn's disease and ulcerative colitis to start the respective treatment early. Perhaps even more important, these extraintestinal symptoms can be the primary manifestation of Crohn's disease and ulcerative colitis. Therefore, they have to be recognized as extraintestinal manifestations to adequately treat the intestinal disease.

Arthritis, Rheumatoid↗
PubMed2004

Optimising the diagnostic value of anti-Saccharomyces cerevisiae-antibodies (ASCA) in Crohn's disease.

BACKGROUND/AIMS: Anti-Saccharomyces cerevisiae-antibodies (ASCA) are used to discriminate Crohn's disease (CD) from ulcerative colitis (UC). ASCA tests are not standardised and different methods for ASCA detection exist. This study was undertaken to compare ASCA tests and to clarify their diagnostic value. PATIENTS/METHODS: One hundred and two sera from CD patients, 53 from UC patients, and 50 sera from normal controls were examined for ASCA IgA and ASCA IgG using four different ELISA (Aesku.lab, Inova, Euroimmun, and Medipan) and indirect immunofluorescence (Euroimmun). Sensitivity, specificity, and positive and negative predictive values (PPV and NPV) for CD in this population were determined. Agreement between tests was expressed by kappa statistics. RESULTS/FINDINGS: The presence of either ASCA IgA or ASCA IgG had a PPV between 77 and 88%. Only the combined presence of ASCA IgA and ASCA IgG had a specificity which was constantly above 90%. Sensitivity, specificity, PPV, and NPV were then: Inova: 53, 97, 95, and 68%; Euroimmun ELISA: 46, 97, 94, and 66%; Aesku.lab: 50, 97, 94, and 66%; Medipan: 30, 98, 94, and 59%; indirect immunofluorescence: 51, 97, 94, and 66%. Agreement between ELISA test results of the Aesku.lab, Euroimmun, Medipan IgA and Inova systems was good (kappa: 0.63-0.79); between the Medipan IgG ELISA or indirect immunofluorescence and the others, it was lower (kappa: 0.33-0.6). If both ASCA IgA and ASCA IgG were detected by indirect immunofluorescence as well as ELISA, specificity for CD increased to >99% at a sensitivity of 23-38%. INTERPRETATION/CONCLUSIONS: The combined detection of ASCA IgA and ASCA IgG by indirect immunofluorescence as well as ELISA may optimise the discrimination of CD from UC.

Adult↗
PubMed2004

Abdominal MRI after enteroclysis or with oral contrast in patients with suspected or proven Crohn's disease.

BACKGROUND & AIMS: Diagnostic results of magnetic resonance (MR) enteroclysis correlate highly with those from conventional enteroclysis; nevertheless, intubation of the patient and positioning of an intestinal tube is still necessary for the examination, which is often remembered as the most embarrassing part of the examination by the patient. A more comfortable and highly sensitive examination of the small bowel therefore would increase patient acceptance for recurring examinations, which are often necessary, for example, in patients with Crohn's disease. This study evaluates the diagnostic efficacy of abdominal MR imaging (MRI) of the small bowel after drinking contrast agent only compared with conventional enteroclysis and abdominal MRI performed after enteroclysis in patients with suspected or proven Crohn's disease. METHODS: Twenty-one patients with Crohn's disease referred for conventional enteroclysis underwent abdominal MRI after enteroclysis. Additionally, 1 to 3 days before or after these examinations, abdominal MRI was performed using only orally administered contrast. All MRI examinations were performed using a 1.5T scanner. RESULTS: All pathological findings on conventional enteroclysis were shown correctly with MRI after enteroclysis and MRI after oral contrast only. Additional information by MRI was obtained in 6 of 21 patients. No statistically significant differences were found in assessing the diagnostic efficacy of the 3 examinations. CONCLUSIONS: Abdominal MRI with oral contrast only can be used as a diagnostic tool for evaluation of the small bowel in patients with Crohn's disease and has the potential to replace conventional enteroclysis as follow-up.

Administration, Oral↗
PubMed2004

Update in inflammatory bowel disease pathogenesis.

PURPOSE OF REVIEW: During the last few years, significant advances have been achieved in the understanding of the pathogenesis of inflammatory bowel disease (IBD). By gaining new insights, paradigms that seemed to be a safe basis of our knowledge on IBD pathogenesis have recently become doubtful. This review discusses and summarizes the most recent developments. RECENT FINDINGS: Important new insights have been gained into the function of caspase-activating and recruitment domain-15 (CARD15)/NOD2, the first cloned susceptibility gene for Crohn disease (CD). New data on CARD15/NOD2 function and nuclear factor-kappaB activation indicate that an inflammatory reaction of the intestinal mucosa as a response of the innate immune system may be necessary for the maintenance of gut homeostasis. CD may therefore be seen as a defective immune response, no longer only as hyperresponsiveness of the mucosal immune system. Data on CARD15/NOD2 expression suggest that macrophages and epithelial cells could be the site of a primary pathophysiologic defect, and T-cell activation might just be a secondary effect inducing chronification of the inflammation, perhaps as a backup mechanism to a defective innate immunity. In addition to CARD15/NOD2, there are additional "innate" pathways by which commensal and pathogenic bacteria can directly interact with cells of the intestinal mucosa (eg, toll-like receptors). The "germ concept" and the "genetic concept" of IBD pathophysiology are converging. SUMMARY: New findings are changing our concepts of the pathogenesis of IBD. The innate immune system, early responses to bacterial products, and the modulation of T-cell responses are important aspects that are reviewed.

Journal Article↗
PubMed2004

Modern imaging using computer tomography and magnetic resonance imaging for inflammatory bowel disease (IBD) AU1.

Radiologic imaging--especially of the small bowel--plays an important role in the diagnosis and management of patients with inflammatory bowel disease. The radiographic examination of the small intestine with barium either as enteroclysis or as small bowel follow through are still the mainstays in small bowel imaging. However, abdominal CT or MRI, which has the advantage of not utilizing ionizing radiation, or the techniques of CT- or MR-enteroclysis, are overall comparable with regard to the sensitivity and specificity in detecting intestinal pathologies and have already replaced the conventional techniques in centers dedicated to the management of inflammatory bowel disease. Additionally, these cross-sectional imaging techniques provide, in a sense, a "one stop abdominal imaging workup," the diagnosis of extraluminal disease manifestations or complications. Future developments of CT- or MR-based virtual colonography and endoscopy in patients with inflammatory bowel disease are currently being investigated, but should momentarily be considered as purely experimental approaches.

Colonoscopy↗
PubMed2004

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 microg/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 microg/ml) diluted in nonconditioned medium induced migration of CLPF in a dose-dependent manner. Maximum migration was induced with 25 microg/ml fibronectin (37 +/- 5 cells/hpf). CONCLUSION: 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.

Autocrine Communication↗
PubMed2003

Functional expression of the interleukin-11 receptor alpha-chain and evidence of antiapoptotic effects in human colonic epithelial cells.

A tissue-protective effect of interleukin-11 (IL-11) for the intestinal mucosa has been postulated from animal models of inflammatory bowel disease (IBD). Despite the fact that the clinical usefulness of the anti-inflammatory effects of this cytokine is presently investigated in patients with IBD, there are no data available regarding the target cells of IL-11 action and the mechanisms of tissue protection within the human colonic mucosa. IL-11 responsiveness is restricted to cells that express the interleukin-11 receptor alpha-chain (IL-11Ralpha) and an additional signal-transducing subunit (gp130). In this study, we identified the target cells for IL-11 within the human colon with a new IL-11Ralpha monoclonal antibody and investigated the functional expression of the receptor and downstream effects of IL-11-induced signaling. Immunohistochemistry revealed expression of the IL-11Ralpha selectively on colonic epithelial cells. HT-29 and colonic epithelial cells (CEC) constitutively expressed IL-11Ralpha mRNA and protein. Co-expression of the signal-transducing subunit gp130 was also demonstrated. IL-11 induced signaling through triggering activation of the Jak-STAT pathway without inducing anti-inflammatory or proliferative effects in colonic epithelial cells. However, IL-11 stimulation resulted in a dose-dependent tyrosine phosphorylation of Akt, a decreased activation of caspase-9, and a reduced induction of apoptosis in cultured CEC. In HLA-B27 transgenic rats treated with IL-11, a reduction of apoptotic cell numbers was found. This study demonstrates functional expression of the IL-11Ralpha restricted on CEC within the human colonic mucosa. IL-11 induced signaling through triggering activation of the Jak-STAT pathway, without inducing anti-inflammatory or proliferative effects. The beneficial effects of IL-11 therapy are likely to be mediated by CEC via activation of the Akt-survival pathway, mediating antiapoptotic effects to support mucosal integrity.

Animals↗
PubMed2003

Stimulation of human colonic epithelial cells by leukemia inhibitory factor is dependent on collagen-embedded fibroblasts in organotypic culture.

The colonic epithelium undergoes a continuous cycle of proliferation, differentiation, and apoptosis. To characterize factors important for colonic homeostasis and its dysregulation, human fetal colonic epithelial cells were isolated and seeded on a collagen type I matrix with embedded colonic fibroblasts. The epithelial cells rapidly spread from clusters and proliferated, and within 3 days, a columnar layer of polarized epithelium surrounded the surface of the constricted collagen matrix. The polarized enterocytes developed brush borders, tight junctions and desmosomes, and goblet and enteroendocrine cells were present. A balance of growth and differentiation was maintained for several weeks in the presence of collagen-embedded fibroblasts and a complex mixture of growth factors. Leukemia inhibitory factor (LIF) was critical for proliferation of enterocytes and inhibited expression of the differentiation marker carbonic anhydrase II. In the presence of LIF, the relative number of goblet cells remained stable, whereas enteroendocrine relative cell number declined. LIF-stimulated epithelial cells remained dependent on the presence of fibroblasts in the matrix. In combination with stem cell factor and endothelin 3, LIF induced formation of disorganized structures of stratified and semi-stratified cells, suggesting that the homeostatic balance in the normal human colon requires cooperation with differentiation-inducing factors.

Carbonic Anhydrase II↗
PubMed2003

Polymorphism of monocyte chemoattractant protein 1 in Crohn's disease.

BACKGROUND AND AIMS: The chemokine MCP-1 is thought to be important for the recruitment of mononuclear cells and the maintenance of inflammation in inflammatory bowel disease. We investigated whether MCP-1 protein expression is correlated with the degree of mucosal inflammation in patients with Crohn's disease. Furthermore, we studied whether a functional single nucleotide polymorphism (G or A) located in the distal regulatory region of the MCP-1 gene is associated with Crohn's disease and/or its phenotype. PATIENTS AND METHODS: MCP-1 concentration in tissue homogenates was analyzed in mucosal biopsy specimens of 31 patients with Crohn's disease and 48 controls by enzyme-linked immunosorbent assay, and the correlation with an endoscopic macroscopic score was analyzed. In 179 patients with Crohn's disease and 189 controls MCP-1 genotyping was carried out by polymerase chain reaction restriction fragment length polymorphism technique. Subgroup phenotypic analysis was performed according to the Vienna classification. RESULTS: MCP-1 tissue concentrations were significantly associated with the macroscopic degree of inflammation. The gene frequency of the different MCP-1 alleles did not differ from healthy controls. However, the G/A and G/G genotype was significantly decreased in patients with a later onset of the disease and both genotypes presented also less frequently with a fistulizing disease behavior. CONCLUSION: The degree of intestinal inflammation in Crohn's disease is associated with MCP-1 tissue levels. Furthermore there is evidence for an association of different disease behavior with different MCP-1 genotypes.

Adult↗
PubMed2003

Reduced migration of fibroblasts in inflammatory bowel disease: role of inflammatory mediators and focal adhesion kinase.

BACKGROUND & AIMS: Crohn's disease (CD) and ulcerative colitis (UC) are associated with chronic tissue damage and continuous tissue repair. A central, but not well-characterized, event during this process is the migration of activated fibroblasts to the wound. METHODS: Human colonic lamina propria fibroblasts (CLPF) were isolated from patients with CD and UC and from healthy controls and were characterized by immunocytochemistry. Migration assays of CLPF were performed in the modified 48-well Boyden chamber. Focal adhesion kinase (FAK) and FAK autophosphorylation in migrating CLPF were determined by Western blotting. FAK mRNA expression was investigated by Northern blotting. RESULTS: The migration of CD-CLPF and UC-CLPF was significantly reduced when compared with control-CLPF. This was correlated with a decrease in FAK phosphorylation, whereas, in migrating control-CLPF, an increase was found. Similarly, the presence of the inflammatory mediators interferon (IFN)-gamma (50 ng/mL) or tumor necrosis factor (TNF) (30 ng/mL) in conditioned medium significantly reduced the migration of control-CLPF to 41% +/- 4% or 30% +/- 7%, respectively. Preincubation of control-CLPF with TNF (20 ng/mL) and IFN-gamma (10 ng/mL) for 3 days reduced their migratory response to 10% of control (P < 0.001), which also was correlated with a decrease in FAK phosphorylation. Culture of IFN-gamma/TNF-treated CLPF for a further 7 days without cytokines did not restore the migratory potential and FAK phosphorylation, indicating a persistent functional change. CONCLUSIONS: CD- and UC-CLPF have a reduced migratory potential compared with normal CLPF. That may be caused by contact with IFN-gamma and TNF. This loss of migratory potential was correlated with diminished FAK phosphorylation.

Adult↗
PubMed2003

Progress on isolation and short-term ex-vivo culture of highly purified non-apoptotic human intestinal epithelial cells (IEC).

Intestinal epithelial cells (IEC) form the largest surface of the human body and are of pivotal importance to digest and absorb nutrients. Furthermore these cells play a critical role shielding the organism against microorganisms and toxins present in the intestinal lumen. It is therefore not surprising that a large group of researchers take great interest in the study of these cells. However, to date it is a challenge to purify viable primary human intestinal epithelial cells and it has been even more fastidious to maintain IEC in culture ex-vivo as IEC undergo apoptosis within hours due to loss of cell anchorage ('anoikis') following the isolation process. Over recent years the authors aimed to continuously improve the isolation technique for primary IEC, allowing a simple, effective and rapid isolation of highly purified non-apoptotic human IEC. In this study the newly improved method is presented and applied to establish ex-vivo cultures of highly purified, fully viable primary IEC displaying important functional properties, making these cells amenable for ex-vivo research on primary human intestinal epithelial cells.

Apoptosis↗