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  • 1
    Keywords: EXPRESSION ; IN-VITRO ; Germany ; TYROSINE KINASE ; DIAGNOSIS ; NETWORKS ; SYSTEM ; TOOL ; DISEASE ; DISEASES ; PROTEIN ; PROTEINS ; TISSUE ; COMPLEX ; COMPLEXES ; RAT ; RATS ; PHOSPHORYLATION ; NERVOUS-SYSTEM ; IDENTIFICATION ; PATTERNS ; NUMBER ; mass spectrometry ; MASS-SPECTROMETRY ; DIETARY ; CENTRAL-NERVOUS-SYSTEM ; HABITS ; protein expression ; POLYACRYLAMIDE GELS ; GEL-ELECTROPHORESIS ; MASSES ; development ; PART ; MASS ; 2-DE ; ADULT-RAT ; enteric nervous system ; ENTERIC NERVOUS-SYSTEM ; MALDI-TOF mass spectrometry ; myenteric plexus ; peripheral nervous system ; PHOSPHOPROTEIN ; Sprague Dawley rats ; STATHMIN ; two-dimensional gel electrophoresis (2-DE)
    Abstract: The enteric nervous system in vertebrates is the most complex part of the peripheral nervous system. Concerning chemical coding, ultrastructure and neuronal circuits, it is more similar to the central than to the peripheral nervous system. Its networks, the myenteric and submucous plexus are integrated in the gut wall. The enteric nervous system is a system of high plasticity, which not only changes during pre- and postnatal development, but also with disease or changing dietary habits. The Aim of this study was to elucidate changes in protein expression during the first two postnatal weeks in the rat myenteric plexus. Colonic and duodenal myenteric plexus from newborn (P1) and fourteen-day old (P14) Sprague-Dawley rats was isolated following a procedure that combines enzymatic digestion and mechanical agitation. The neuronal tissue was collected and processed for two-dimensional gel electrophoresis (2-DE). The obtained 2-D gels were stained with silver for image analysis or with colloidal Coomassie for subsequent protein identification. Gels from the various samples showed a high degree of consistence concerning protein-spots found in all preparations. Nevertheless, there was a number of proteins that were clearly detected in one sample but not, or only in significantly smaller amounts in the other. Several differentially expressed proteins in the postnatal myenteric plexus were identified with MALDI-TOF mass spectrometry. Especially stathmin, polyubiquitin and heterogeneous nuclear ribonucleoprotein seem to play an important role in pre- and postnatal development. 2-DE combined with mass spectrometry can help to identify pathological relevant proteins in the enteric nervous system, and so deliver a valuable tool for the early diagnosis of also central nervous system diseases by using biopsies from the gut. (c) 2005 Elsevier B.V All rights reserved
    Type of Publication: Journal article published
    PubMed ID: 16183334
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  • 2
    Keywords: CELLS ; EXPRESSION ; IN-VITRO ; tumor ; carcinoma ; CELL ; Germany ; MODEL ; VITRO ; GENE ; GENES ; HYBRIDIZATION ; PROTEIN ; PROTEINS ; SAMPLES ; TUMORS ; MARKER ; prognosis ; CARCINOGENESIS ; SEQUENCE ; SUPPRESSION ; ALPHA ; MOUSE ; COMPARATIVE GENOMIC HYBRIDIZATION ; CANCER-CELLS ; TRANSFORMATION ; adenocarcinoma ; ki-ras ; K-RAS ; HUMAN BREAST-TUMORS ; MOLECULAR-CLONING ; PROTEOMICS ; Ras ; PROTEOMIC ANALYSIS ; 2-DIMENSIONAL GEL-ELECTROPHORESIS ; DRUG-INDUCED APOPTOSIS ; RE ; TRANSFECTION ; development ; pancreatic ; BOVINE ; UNIT ; BINDING PROTEINS ; pancreatic carcinogenesis ; ANNEXIN-I ; SV40 large T ; transcriptomics
    Abstract: Pancreatic carcinoma has an extremely bad prognosis due to lack of early diagnostic markers and lack of effective therapeutic strategies. Recently, we have established an in vitro model recapitulating the first steps in the carcinogenesis of the pancreas. SV40 large T antigen-immortalized bovine pancreatic duct cells formed intrapancreatic adenocarcinoma tumors on k-ras(mut) transfection after orthotopic injection in the nude mouse pancreas. Here we identified genes and proteins differentially expressed in the course of malignant transformation using reciprocal suppression subtractive hybridization and 2D gel electrophoresis and mass spectrometry, respectively. We identified 34 differentially expressed genes, expressed sequence tags, and 15 unique proteins. Differential expression was verified for some of the genes or proteins in samples from pancreatic carcinoma. Among these genes and proteins, the majority had already been described either to be influenced by a mutated ras or to be differentially expressed in pancreatic adenocarcinoma, thus proving the feasibility of our model. Other genes and proteins (e.g., BBC1, GLTSCR2, and rhoGDI alpha), up to now, have not been implicated in pancreatic tumor development. Thus, we were able to establish an in vitro model of pancreatic carcinogenesis, which enabled us to identify genes and proteins differentially expressed during the early steps of malignant transformation
    Type of Publication: Journal article published
    PubMed ID: 17356710
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  • 3
    Keywords: INHIBITOR ; Germany ; MICROSCOPY ; THERAPY ; PROTEIN ; PROTEINS ; COMPONENTS ; PATIENT ; MECHANISM ; mechanisms ; IDENTIFICATION ; mass spectrometry ; MASS-SPECTROMETRY ; LENGTH ; Jun ; DEGRADATION ; CALCIUM ; PROTEASOME ; chronic pancreatitis ; MANAGEMENT ; GEL-ELECTROPHORESIS ; INHIBITORS ; MASSES ; albumin ; PRODUCTS ; LIGHT ; PH ; WEIGHT ; SODIUM ; development ; methods ; pancreatic ; MASS ; PLACEMENT ; CRITERIA ; CRYSTAL ; JUICE ; LITHOSTATHINE ; STENTS ; STONE PROTEIN ; SULFATE ; technique
    Abstract: Background: Endoscopic management of chronic pancreatitis (CP), especially pancreatic stent placement, has made tremendous advances. However, good clinical results are hampered by rapid occlusion. The objective of this study was to understand mechanisms and materials that cause stent occlusion. Methods: The clogging material of 50 lyophilized pancreatic endoprostheses (length 8.5 cm, range 5-14 cm, diameter 7-11F) from patients with CP was completely removed and weighed. Protein solubilization was achieved at pH 8.0 by using sodium dodecyl sulfate (SDS) and 2-mercaptoethanol in the presence of proteasome inhibitors. Proteins were separated by using a SDS-polyacrylamide gel electrophoresis. Protein identification was performed by the Western blot technique, as well as by mass spectrometry. Insoluble components were examined by polarized light microscopy and after staining (periodic acid-Schiff [PAS]). Results: Clogging material was found in 49 prostheses, mainly at the duodenal flap (80%). More than a third of the prostheses contained visible calcium carbonate calculi. Light microscopy and PAS staining showed plant debris (80%), crystals (73.5%), and mucopolysaccharides (100%). The dry weight of clogging material (18 +/- 13 mg, range 3-72 mg) correlated significantly with the stent diameter (p = 0.029) but not with any other stent- or patient-related criteria. Albumin, its degradation products, and lithostathine were identified as the main proteinaceous components. Conclusions: Almost all pancreatic stents had clogging material, predominantly located at the duodenal flap, which contained plant material, mucopolysaccharides, and crystals, as well as visible calcium carbonate calculi. Albumin and lithostathine may play an important role in the development of stent occlusion
    Type of Publication: Journal article published
    PubMed ID: 15933688
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