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1.
Can J Surg ; 55(6): 389-93, 2012 Dec.
Article in English | MEDLINE | ID: mdl-22992401

ABSTRACT

BACKGROUND: Different approaches to study liver regeneration in murine models have been proposed. We investigated the effect of different liver resection models on liver damage and regeneration parameters in mice. METHODS: We compared the technical aspect of the 2 most commonly used techniques of 50% and 70% liver resection. Liver damage, as determined by the change in serum alanine aminotransferase and aspartate aminotransferase, as well as the regeneration parameters VEGF and FGF-2 were analyzed at 6 time points. A postoperative vitality score was introduced. RESULTS: Cholestasis was not observed for either technique. Both resection techniques resulted in full weight recovery of the liver after 240 hours, with no significant difference between sham and resection groups. Postoperative animal morbidity and total protein levels did not differ significantly for either method, indicating early and full functional recovery. However, comparing the mitogenic growth factors FGF-2 and VEGF, a significant increase in serum levels and, therefore, increased growth stimulus, was shown in the extended resection group. CONCLUSION: Extended resection led to a greater response in growth factor expression. This finding is important since it shows that growth factor response differs acdording to the extent of resection. We have demonstrated the need to standardize murine hepatic resection models to adequately compare the resulting liver damage.


Subject(s)
Fibroblast Growth Factor 2/metabolism , Hepatectomy/adverse effects , Hepatectomy/methods , Liver Regeneration , Liver/injuries , Liver/metabolism , Vascular Endothelial Growth Factor A/metabolism , Alanine Transaminase/blood , Animals , Aspartate Aminotransferases/blood , Bilirubin/blood , Biomarkers/blood , Blood Proteins/metabolism , Liver/enzymology , Liver Diseases/metabolism , Liver Diseases/surgery , Male , Mice , Mice, Inbred BALB C , Movement
2.
Anticancer Res ; 30(12): 5257-61, 2010 Dec.
Article in English | MEDLINE | ID: mdl-21187523

ABSTRACT

BACKGROUND: To date, EGF 61*A/G, TGF-ß1 -509*T/C and TNF-α -308*A/G gene polymorphisms have been not been analysed in pancreatic carcinoma. This study investigated the frequency of these gene polymorphisms among patients with cancer of the pancreatic head. PATIENTS AND METHODS: A total of 73 pancreatic head cancer patients and 117 cancer-free healthy people were recruited at the Surgical Department of the University Hospital Mannheim. Genomic DNA was isolated from peripheral blood and gene polymorphisms were analysed by PCR-RFLP. RESULTS: The distribution of EGF 61*G/G homozygotes among pancreatic head cancer patients was more frequent than that in the control group (24.7% vs 11.1%, odds ratio (OR) = 2.618, 95% confidence interval (CI) = 1.195-5.738). In addition, the frequency of the G allele in the pancreatic head cancer patient group was also higher than that in the control group (45.9% vs. 33.3%, OR = 1.696, 95% CI = 1.110-2.592). No difference was found for the TGF-ß1 -509 and TNF-α -308 genotypes among pancreatic head cancer patients and healthy controls. CONCLUSION: The frequencies of the EGF 61*G/G genotype and G allele are significantly increased among patients with pancreatic head cancer. TGF-ß1-509*T/C and TNF-α -308*A/G gene polymorphisms are not related to this cancer entity.


Subject(s)
Epidermal Growth Factor/genetics , Pancreatic Neoplasms/genetics , Transforming Growth Factor beta1/genetics , Tumor Necrosis Factor-alpha/genetics , Adult , Aged , Aged, 80 and over , Case-Control Studies , Female , Humans , Male , Middle Aged , Polymerase Chain Reaction , Polymorphism, Single Nucleotide
3.
World J Surg ; 33(1): 124-9, 2009 Jan.
Article in English | MEDLINE | ID: mdl-19011936

ABSTRACT

INTRODUCTION: Up to the present, EGF 61 A/G, TGF-beta1 -509 T/C, and VEGF 936 T/C gene polymorphisms have been analyzed in other cancer entities than colorectal cancer. We have now investigated the frequency of these gene polymorphisms among colorectal cancer patients. MATERIAL AND METHODS: A total of 157 colorectal cancer patients and 117 cancer-free healthy people were recruited at the Surgical Department of the Universitätsklinikum Mannheim. All patients and healthy people are Caucasians. Genomic DNA was isolated from peripheral blood, and gene polymorphisms were analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). RESULTS: The distribution of EGF 61 G/G homozygotes among colorectal cancer patients was more frequent than that in the control group (33.1% versus 11.1%; Odds Ratio [OR]=3.962; 95% Confidence Interval [CI]=2.036-7.708). The frequency of the "G" allele in the colorectal cancer patient group was also higher than that in the control group (51.3% versus 33.3%; OR=2.105; 95% CI=1.482-2.988). No difference could be found for the TGF-beta1 and VEGF genotypes among colorectal cancer patients and healthy controls. CONCLUSIONS: The EGF 61 G/G genotype and the G allele are significantly related to colorectal cancer. The TGF-beta1 -509 T/C and VEGF 936 T/C gene polymorphisms are not related to colorectal cancer.


Subject(s)
Colorectal Neoplasms/genetics , Epidermal Growth Factor/genetics , Polymorphism, Genetic/genetics , Transforming Growth Factor beta1/genetics , Vascular Endothelial Growth Factor A/genetics , Adult , Aged , Aged, 80 and over , Female , Gene Frequency/genetics , Genetic Predisposition to Disease , Genotype , Humans , Male , Middle Aged
4.
Surg Laparosc Endosc Percutan Tech ; 18(6): 561-4, 2008 Dec.
Article in English | MEDLINE | ID: mdl-19098660

ABSTRACT

Perforations of the colonic wall or splenic injury during colonoscopy are rare complications. Treatment of these complications by laparoscopy is an advisable compromise instead of an invasive surgery with a laparotomy or a noninvasive and potentially risky nonoperative therapy. All surgical procedures that can be performed by open approach can also be performed laparoscopically. We present in this report 15 patients who were treated for a perforation after colonoscopy. In addition, 2 cases of splenic injury after colonoscopy are described. Twelve perforations were sutured laparoscopically and 3 perforations were sutured via laparotomy. Except for 1 minor wound infection, there were no complications. One splenic injury was treated by spleen wrapping via an open approach due to former pancreatic surgery, and 1 injury was treated laparoscopically with a hemostypticum. Mortality was 0%. Early laparoscopic intervention is a safe and effective method in the treatment of serious complications after colonoscopy.


Subject(s)
Colonoscopy/adverse effects , Digestive System Surgical Procedures/methods , Intestinal Perforation/etiology , Intestinal Perforation/surgery , Laparoscopy/methods , Aged , Colon/injuries , Colon/surgery , Female , Humans , Length of Stay , Middle Aged , Spleen/injuries , Spleen/surgery , Splenic Diseases/etiology , Splenic Diseases/surgery , Treatment Outcome
5.
Zhonghua Wei Chang Wai Ke Za Zhi ; 11(6): 569-71, 2008 Nov.
Article in Chinese | MEDLINE | ID: mdl-19031138

ABSTRACT

OBJECTIVE: To elucidate the association of TNF-alpha-308G/A gene functional polymorphism with the development and progression of colorectal cancer. METHODS: PCR-RFLP was employed to detect the TNF-alpha-308 G/A genotypes in 157 colorectal cancer patients and 117 healthy controls. RESULTS: The frequency of TNF-alpha-308 genotype and allele were not significantly different between colorectal cancer patients and healthy controls (genotype chi(2)=1.054, P=0.591, allele chi(2)=0.404, P=0.525). The frequency of A/A genotype and A allele in III+IV stage (62 patients in total) were higher than those in I+II stages (85 patients in total) (A allele: 22.6% vs 12.9%, A/A genotype: 8.1% vs 1.2%), and the differences were significant (genotype P=0.048, OR=7.368, 95% CI=0.839-64.743, allele chi(2)=4.720, P=0.03, OR=1.962, 95% CI=1.061-3.628). The frequency of TNF-alpha-308 genotype were not significantly different among different colorectal cancer grades (chi(2)=3.009,P=0.591). CONCLUSION: TNF-alpha-308G/A gene polymorphism is not associated with the development of colorectal cancer, but TNF-alpha-308 A/A genotype and A allele are related to the progression of colorectal cancer.


Subject(s)
Colorectal Neoplasms/genetics , Polymorphism, Single Nucleotide , Tumor Necrosis Factor-alpha/genetics , Adult , Aged , Aged, 80 and over , Alleles , Case-Control Studies , Colorectal Neoplasms/pathology , Female , Gene Frequency , Genetic Predisposition to Disease , Genotype , Humans , Male , Middle Aged , Neoplasm Staging
7.
Zhonghua Wai Ke Za Zhi ; 44(21): 1505-7, 2006 Nov 01.
Article in Chinese | MEDLINE | ID: mdl-17349182

ABSTRACT

OBJECTIVE: To investigate the association between VEGF gene 936 T/C polymorphism and colorectal cancer together with anastomotic leakage. METHODS: Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to detect the VEGF 936 T/C genotypes in colorectal cancer patients and healthy controls. RESULTS: There was no significant difference in the frequency of VEGF 936 C/C genotype or C allele between colorectal cancer patients and healthy controls (P > 0.05). The C/C genotype or C allele in colorectal cancer patients with anastomotic leakage was less frequently found than in the group without anastomotic leakage (P < 0.05). CONCLUSIONS: VEGF 936 C/C genotype or C allele is not related to the development of colorectal cancer, but they can reduce the risk of anastomotic leakage after surgery in colorectal cancer patients.


Subject(s)
Colorectal Neoplasms/genetics , Digestive System Fistula/genetics , Endothelial Growth Factors/genetics , Polymorphism, Single Nucleotide , Adult , Aged , Aged, 80 and over , Alleles , Colorectal Neoplasms/surgery , Digestive System Fistula/etiology , Female , Gene Frequency , Genotype , Humans , Male , Middle Aged , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Postoperative Complications/etiology , Surgical Stomas/adverse effects
8.
J Biol Chem ; 280(30): 27826-31, 2005 Jul 29.
Article in English | MEDLINE | ID: mdl-15908435

ABSTRACT

In tumor cells, high phosphorylation levels of receptor tyrosine kinases may occur in the absence of exogenous ligands due to autocrine signaling or enhanced tyrosine kinase activity. Here we show that the phosphorylation state of the endogenous epidermal growth factor receptor (EGFR) can be quantitatively imaged in tumor cells and tissues by detecting fluorescence resonance energy transfer between fluorophores conjugated to antibodies against the receptor and phosphotyrosine, respectively. Five different human colorectal cell lines were analyzed for activity and expression of EGFR. All cell lines exhibited basal EGFR phosphorylation under serum starvation conditions. Phosphorylation levels increased after stimulation with EGF or pervanadate, dependent on the level of basal EGFR phosphorylation in the respective cell lines. This basal activity correlated inversely with receptor expression. Using the acceptor photobleaching fluorescence resonance energy transfer imaging approach, a significantly higher phosphorylation state of EGFR was also found in resected human colorectal tumor samples as compared with adjacent healthy tissue. Imaging of EGFR phosphorylation may thus serve as a valuable tool to investigate the role of receptor tyrosine kinase activity in malignant cell growth.


Subject(s)
Colorectal Neoplasms/metabolism , ErbB Receptors/chemistry , Fluorescence Resonance Energy Transfer/methods , Microscopy, Confocal/methods , Cell Line, Tumor , Cell Proliferation , Cloning, Molecular , Culture Media, Serum-Free/pharmacology , DNA, Complementary/metabolism , Enzyme-Linked Immunosorbent Assay , ErbB Receptors/metabolism , Humans , Immunoblotting , Immunoprecipitation , Kinetics , Microscopy, Fluorescence , Phosphorylation , Phosphotyrosine/metabolism , RNA/metabolism , Receptor Protein-Tyrosine Kinases/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Signal Transduction , Time Factors , Vanadates/pharmacology
9.
Pancreas ; 29(3): 204-11, 2004 Oct.
Article in English | MEDLINE | ID: mdl-15367886

ABSTRACT

OBJECTIVES: The cysteine proteases cathepsin B (CTSB) and L (CTSL) have been implicated in tumor spread and metastatic formation. In pancreatic adenocarcinoma, the role of these proteases is not very well defined. To find out which cell types produce CTSB and CTSL and to evaluate the prognostic impact of these proteases, 70 specimens from curatively resected patients with pancreatic adenocarcinoma were examined by in situ hybridization and immunohisto-chemistry. METHODS: Seventy patients with ductal adenocarcinoma of the pancreas were studied after R0 resection with a follow-up of at least 3 years. CTSB and CTSL expression was performed immunohisto-chemically using polyclonal anti-CTSB and CTSL antibodies. To detect cell types involved in producing CTSB and CTSL as well as the intracellular localization of specific mRNA sequences, nonisotopic in situ hybridization was performed. The correlations among CTSB and CTSL expression, clinicopathologic parameters, and clinical outcome were analyzed. RESULTS: The immunoreactivity was 96% for CTSB and 90% for CTSL. Positive mRNA signals were obtained in the cytoplasm tumor cells, macrophages, and fibroblasts in 77% for CTSB and 81% for CTSL, respectively. Statistical analysis showed a significant correlation between CTSB/CTSL expression and tumor grading (P < 0.05) and between CTSB and lymphatic invasion (P = 0.05). Kaplan-Meier analyses revealed statistical significance for CTSB/CTSL expression with the survival after curative resection (P < 0.05). Both proteases are strong prognostic markers in multivariate analysis (P = 0.0001) beside UICC stage, nodal status, tumor size, and grading (P < 0.05). Furthermore, CTSB expression is an independent prognostic marker for cancer recurrence within 6 months after curative surgery in multivariate analysis (P = 0.0001). CONCLUSIONS: CTSB and CTSL are strong and independent prognostic markers in resectable pancreatic adenocarcinoma rather than UICC stage, TNM classification, or tumor grading. Furthermore, CTSB is a predictor for early recurrence after curative resection. These data underline the significance of tumor-associated proteolysis for cancer invasion and metastasis and may lead to defining subgroups of patients with early recurrence and poor outcome.


Subject(s)
Biomarkers, Tumor/analysis , Carcinoma, Pancreatic Ductal/enzymology , Cathepsin B/analysis , Cathepsins/analysis , Cysteine Endopeptidases/analysis , Neoplasm Proteins/analysis , Pancreatic Neoplasms/enzymology , Adult , Aged , Biomarkers, Tumor/biosynthesis , Carcinoma, Pancreatic Ductal/mortality , Carcinoma, Pancreatic Ductal/pathology , Carcinoma, Pancreatic Ductal/secondary , Carcinoma, Pancreatic Ductal/therapy , Cathepsin B/biosynthesis , Cathepsin L , Cathepsins/biosynthesis , Combined Modality Therapy , Cysteine Endopeptidases/biosynthesis , Endothelial Cells/chemistry , Epithelial Cells/chemistry , Female , Fibroblasts/chemistry , Follow-Up Studies , Humans , In Situ Hybridization , Islets of Langerhans/chemistry , Life Tables , Liver Neoplasms/secondary , Macrophages/chemistry , Male , Middle Aged , Neoplasm Invasiveness , Neoplasm Proteins/biosynthesis , Pancreatic Neoplasms/mortality , Pancreatic Neoplasms/pathology , Pancreatic Neoplasms/therapy , Prognosis , RNA, Messenger/analysis , RNA, Neoplasm/analysis , Subcellular Fractions/enzymology , Survival Analysis
10.
Cell Physiol Biochem ; 14(4-6): 249-60, 2004.
Article in English | MEDLINE | ID: mdl-15319528

ABSTRACT

BACKGROUND/AIMS: To investigate postulated differences related to FGF-2 in liver morphology and expression of apoptosis-related factors after partial hepatectomy (PH). METHODS: Homogenous FGF-2-deficient mice (C57BL/6J) with their FGF-2-(+/+) littermates (control) were used to examine the structure of regenerating livers after PH with light and electron microscopy. The regenerative response and BrDu incorporation were monitored. The expression of BclX-l, Bax, Fas, TNF-alpha, and Caspase-3 were measured by reverse transcription PCR (RT-PCR) and Northern blot analysis. RESULTS: In the FGF-2-(-/-) group, hepatocytes and endothelial cells contain mitochondria with atypical cristae and fragmented endoplasmic reticulum structures compared to control. Sinusoids show irregular basal laminae. These changes are in accordance with a differential expression of apoptosis-related factors: FasL was expressed throughout the entire observation span (days 0 to 10 post-PH). Following PH, tumor necrosis factor alpha (TNFalpha)-mRNA levels were higher in FGF-2-(+/+) animals, while Fas as well as Bax and BclXl were overexpressed in FGF-2-(-/-) mice. Caspase-3-mRNA was similarly expressed in both groups, but Caspase-3 activity was elevated for 4 days in FGF-2-(-/-) mice. CONCLUSION: Despite morphologic differences, differences in the time schedule of DNA synthesis and differences in apoptotic response, the dynamics of liver regeneration in FGF-2-(-/-) mice were not impaired.


Subject(s)
Apoptosis/physiology , Fibroblast Growth Factor 2/genetics , Hepatocytes/ultrastructure , Liver Regeneration/physiology , Animals , Apoptosis/genetics , Caspase 3 , Caspases/analysis , Caspases/genetics , Caspases/metabolism , Cell Size , Endoplasmic Reticulum/ultrastructure , Fas Ligand Protein , Fibroblast Growth Factor 2/physiology , Gene Expression , Hepatectomy , Hepatocytes/chemistry , Hepatocytes/physiology , Liver/ultrastructure , Liver Regeneration/genetics , Membrane Glycoproteins/genetics , Membrane Glycoproteins/metabolism , Mice , Mice, Mutant Strains , Mitochondria/ultrastructure , Proto-Oncogene Proteins c-bcl-2/genetics , Proto-Oncogene Proteins c-bcl-2/metabolism , RNA, Messenger/analysis , RNA, Messenger/metabolism , Tumor Necrosis Factor-alpha/genetics , Tumor Necrosis Factor-alpha/metabolism , bcl-2-Associated X Protein , bcl-X Protein , fas Receptor/genetics , fas Receptor/metabolism
11.
Cancer Lett ; 208(2): 197-206, 2004 May 28.
Article in English | MEDLINE | ID: mdl-15142679

ABSTRACT

The expression of 14 individual and two groups of tumor antigens was characterized for colorectal carcinoma by RT-PCR using 26 colorectal carcinoma specimens, eight cell lines, six samples of patients with inflammatory bowl diseases, and nine specimens from different locations of an individual patient with a metastasized rectal carcinoma. The most frequently detected mRNAs were MAGE-A1 (58%), GAGE-3-7 (54%), and cTAGE-5a (31%). At medium frequencies (12-19%) we found cTAGE-1, MAGE-A2, se57-1, RAGE-4, and GAGE-1,2,8, while other tumor antigens were expressed rarely (<9%). 85% of the samples were positive for at least one of the most frequently expressed antigens. Using a secondary SEREX approach and sera of eight colorectal cancer patients we found reactive antibodies against recombinant cTAGE-1 (2 sera), se57-1 (2), truncated GAGE (1), and MAGE-A1 (1). We conclude that certain cancer-germline genes can be detected in colorectal cancer and might therefore be promising targets for immunotherapy.


Subject(s)
Antigens, Neoplasm/genetics , Colorectal Neoplasms/immunology , Antigens, Differentiation, T-Lymphocyte , Antigens, Neoplasm/immunology , Humans , Melanoma-Specific Antigens , Membrane Glycoproteins , Neoplasm Proteins/genetics , RNA, Messenger/analysis , Reverse Transcriptase Polymerase Chain Reaction
12.
Liver Int ; 24(2): 161-8, 2004 Apr.
Article in English | MEDLINE | ID: mdl-15078481

ABSTRACT

BACKGROUND/AIMS: The angiogenic properties, its role in mesoderm differentiation and cell culture studies implicate an important role of fibroblast growth factor (FGF-2) in liver regeneration. The aim of the study was to evaluate this role in a FGF-2 knockout mouse model. METHODS: Liver regeneration after left hemihepatectomy (partial hepatectomy, PH) was evaluated in homozygous FGF-2 deficient (-/-) mice (male C57BL/6J) and their FGF-2 competent (+/+) littermates (controls) (day 0-10). RESULTS: FGF-2-(-/-) mice displayed normal dynamics in liver regeneration. FGF-2 protein was overexpressed 4 days post PH in controls. BrdU incorporation showed a biphasic pattern in FGF-2-(-/-) mice, whereas it decreased continuously after one peak (day 2) in controls. In FGF-2-(-/-) livers hepatic growth factor mRNA post PH was 1 day longer decreased and markedly less elevated thereafter compared with control. Vascular endothelial growth factor (VEGF) mRNA levels were clearly increased in FGF-2-(-/-) mice pre- and postoperatively in contrast to controls. VEGF protein levels in livers of FGF-2-(-/-) mice were elevated preoperatively, but similar in both groups after PH. With SU5416, a VEGF-receptor inhibitor, liver regeneration in FGF-2-(-/-) mice was reduced significantly, whereas it remained unchanged in controls. CONCLUSIONS: Liver regeneration dynamics in FGF-2-(-/-) mice were comparable with controls, potentially due to a functional substitution of FGF-2 by VEGF.


Subject(s)
Fibroblast Growth Factor 2/genetics , Fibroblast Growth Factor 2/metabolism , Liver Regeneration/physiology , Vascular Endothelial Growth Factor A/genetics , Vascular Endothelial Growth Factor A/metabolism , Angiogenesis Inhibitors/administration & dosage , Angiogenesis Inhibitors/pharmacology , Animals , Hepatectomy , Homozygote , Indoles/administration & dosage , Indoles/pharmacology , Injections, Intraperitoneal , Liver Regeneration/drug effects , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Neovascularization, Pathologic/metabolism , Pyrroles/administration & dosage , Pyrroles/pharmacology , RNA, Messenger/genetics , RNA, Messenger/metabolism , Vascular Endothelial Growth Factor Receptor-2/antagonists & inhibitors
13.
Clin Exp Metastasis ; 20(5): 395-405, 2003.
Article in English | MEDLINE | ID: mdl-14524528

ABSTRACT

Many animal models for metastatic colorectal cancer represent clinical manifestations just inaccurately. We introduce a novel mouse model for metastastatic colorectal cancer. In order to remain close to the clinical disease a syngenic murine colon carcinoma cell line (colon 26 cells) was transfected with enhanced green fluorescent protein (EGFP). The transfected cells maintain the highly malignant attributes of the wild-type cells. Following injection into the portal circulation of Balb/c-mice, liver metastases occur in the same time span. Using the fluorescent attributes of the transfected cells, an approximation of the tumor load in liver tissue can be achieved by fluorescence activated cell sorting (FACS) and fluoroscan analysis. Tumor cell load in liver tissue can be accurately measured by Northern blot and Western blot analysis of liver tissue containing EGFP-transfected colon cancer metastases (1250 cells/mg liver tissue and 1000 cells/mg liver tissue) respectively. Confocal microscopy and intravital microscopy confirmed the growth of tumor metastases, originating from the intravascular compartments. The presented animal model using EGFP-transfected colon 26 cells allows the detecting of tumor growth in vivo and post mortem, as well as an accurate quantification of the tumor load in the liver tissue.


Subject(s)
Luminescent Proteins/metabolism , Animals , Blotting, Northern , Blotting, Western , Cell Division , Cell Separation , Cloning, Molecular , DNA, Complementary/metabolism , Female , Flow Cytometry , Green Fluorescent Proteins , Hepatocytes/metabolism , Liver/metabolism , Liver/pathology , Mice , Mice, Inbred BALB C , Microscopy, Confocal/methods , Microscopy, Fluorescence , Microscopy, Video , Neoplasm Metastasis , Neoplasm Transplantation , Time Factors , Transfection , Tumor Cells, Cultured
14.
Int J Cancer ; 107(1): 11-21, 2003 Oct 20.
Article in English | MEDLINE | ID: mdl-12925951

ABSTRACT

The level of TNFalpha expression is increased after partial hepatectomy, and experimental evidence exists that TNFalpha plays a key role in liver regeneration. Contradictory results are reported about the influence of TNFalpha on tumor growth: on the one hand, stimulation of tumor growth in various animal models and, on the other hand, intraperitoneally administered TNFalpha leads to reduced metastasis formation. TNFalpha may be one responsible factor for increased metastasis formation after surgical trauma. The objective of our study was to clarify the influence of TNFalpha on the formation of liver metastases in a syngenic mouse model in vivo. We used a novel marker system, EGFP transfected C26 tumor cells for in vivo observation of metastasis formation by intravital microscopy. We analyzed the effect of intraperitoneal TNFalpha-injection on tumor cell adhesion, extravasation and tumor development. The expression of ICAM-1, VCAM-1 and E-Selectin was measured by Western blot and immunohistochemical staining. Tumor load was assessed by determining EGFP in Western blots. GdCl(3) was employed 24 and 48 hr before tumor cell injection to selectively deplete the liver of functioning Kupffer cells. We observed significantly more extravasated tumor cells in the TNFalpha-pre-treated animals at early time points with increased expression of adhesion molecules. Measurement of the EGFP levels showed fewer liver metastases in the TNFalpha-pretreated animals at day 8. After GdCl(3) pretreatment even lower levels of EGFP, i.e., fewer metastases and also lower expression levels of ICAM-1, VCAM-1 and E-Selectin could be observed. TNFalpha, acts in a bidirectional manner: whereas TNFalpha facilitates tumor cell adhesion and extravasation of C26 tumor cells by inducing the expression of adhesion molecules, at later time points, TNFalpha seems to hinder the formation of liver metastases.


Subject(s)
Adenocarcinoma/metabolism , Colonic Neoplasms/metabolism , Liver Neoplasms/metabolism , Liver Neoplasms/secondary , Tumor Necrosis Factor-alpha/metabolism , Adenocarcinoma/pathology , Animals , Anti-Inflammatory Agents/pharmacology , Colonic Neoplasms/pathology , Disease Models, Animal , E-Selectin/metabolism , Fluorescent Dyes , Gadolinium/pharmacology , Green Fluorescent Proteins , Indicators and Reagents/metabolism , Injections, Intravenous , Intercellular Adhesion Molecule-1/metabolism , Luminescent Proteins/metabolism , Male , Mice , Mice, Inbred BALB C , RNA, Messenger/metabolism , Rhodamines , Tumor Cells, Cultured/transplantation , Tumor Necrosis Factor-alpha/genetics , Vascular Cell Adhesion Molecule-1/metabolism
15.
Resuscitation ; 58(2): 219-25, 2003 Aug.
Article in English | MEDLINE | ID: mdl-12909385

ABSTRACT

We report a case of sudden death after gas gangrene. A 67-year-old male patient with diabetes mellitus and chronic renal failure (on haemodialysis three times a week) presented in the surgical emergency department with a severe swelling and crepitation in the right groin. No signs of trauma were present-except for a well-healed, 1-year-old scar after femoro-popliteal bypass surgery. Two days earlier, he had presented to the internal medicine department with epigastric pain and had left against medical advice. On readmission the patient was initially conscious and in a stable cardiopulmonary condition but developed sudden cardiocirculatory failure and underwent resuscitation. Despite all resuscitation measures, including the administration of high doses of catecholamines and the treatment of hyperkalemia, the patient died. Autopsy revealed septicaemia with rod-shaped gram-positive bacteria, typical of Clostridium perfringens, evidenced by multiple areas of myonecrosis. Abscess formation was found in the myocardium. Clostridial gas gangrene is a rare clinical condition. Unless immediate diagnosis and adequate therapy measures are taken, the outcome and chances for survival are poor as demonstrated by this case.


Subject(s)
Abscess/etiology , Cardiomyopathies/etiology , Gas Gangrene/complications , Shock, Septic/etiology , Abscess/pathology , Aged , Cardiomyopathies/pathology , Cardiopulmonary Resuscitation , Gas Gangrene/mortality , Gas Gangrene/pathology , Heart Arrest/therapy , Humans , Male , Shock, Septic/pathology
16.
Shock ; 19(4): 378-82, 2003 Apr.
Article in English | MEDLINE | ID: mdl-12688551

ABSTRACT

In the obstructed gut, nitric oxide (NO) may influence intestinal barrier function and translocation of bacteria. By using a novel experimental approach, we investigated the effect of supplementation and inhibition of NO synthesis on the time interval necessary for translocation of green fluorescent protein-transfected Escherichia coli (GFP-uv E. coli) in a rat model of small bowel obstruction. In anesthetized Wistar rats, 4 x 10(8) GFP-uv E. coli were administered into a reservoir of terminal ileum formed by ligature. Animals were randomized to receive either i.v. arginine (10 mg/kg), aminoguanidine (300 mg/kg), L-NAME (25 mg/kg), or saline (control). Translocation of GFP-uv E. coli was assessed using intravital video microscopy. Minimal transit time of translocation was measured as time from injection of GFP-uv E. coli into the gut lumen until bacteria were observed in the lamina submucosa and as time from injection of bacteria into the gut lumen until bacteria were observed in the lamina muscularis propria. Minimal transit times were expressed as mean +/- SD. Bacterial translocation into the submucosa and muscularis propria took 36 +/- 7 min and 81 +/- 9 min, respectively in control animals receiving saline. Aminoguanidine and L-NAME caused a marked delay of minimal transit time into the submucosa (63 +/- 5 min and 61 +/- 7 min, respectively; P < 0.05). Arginine significantly accelerated bacterial translocation into the muscularis propria (61 +/- 9 min, P < 0.05). GFP-uv E. coli were detected on frozen sections of small bowel, mesentery, liver, and spleen 2 h after GFP-uv E. coli administration in all animals. A marked upregulation of inducible NO synthase (NOS) in the obstructed bowel segment was demonstrated on immunohistochemistry. The assessment of a newly defined parameter, minimal bacterial transit time, may serve as an additional functional aspect of intestinal barrier function for pathophysiological and pharmacological studies. Aminoguanidine, L-NAME, and arginine were effective in influencing minimal transit time of E. coli during small bowel obstruction.


Subject(s)
Arginine/pharmacology , Bacterial Translocation/drug effects , Enzyme Inhibitors/pharmacology , Escherichia coli/physiology , Guanidines/pharmacology , Ileal Diseases/microbiology , Intestinal Obstruction/microbiology , NG-Nitroarginine Methyl Ester/pharmacology , Nitric Oxide Donors/pharmacology , Nitric Oxide/pharmacology , Animals , Bacterial Translocation/physiology , Escherichia coli/chemistry , Genes, Reporter , Green Fluorescent Proteins , Ileal Diseases/complications , Intestinal Mucosa/drug effects , Intestinal Mucosa/microbiology , Intestinal Obstruction/complications , Luminescent Proteins/analysis , Luminescent Proteins/genetics , Male , Models, Animal , Muscle, Smooth/drug effects , Muscle, Smooth/microbiology , Nitric Oxide Synthase/antagonists & inhibitors , Rats , Rats, Wistar , Time Factors , Transfection
17.
World J Surg ; 27(3): 324-9, 2003 Mar.
Article in English | MEDLINE | ID: mdl-12607060

ABSTRACT

The prognosis of patients who undergo resection for pancreatic ductal adenocarcinoma with curative intention is generally poor unless they have early-stage disease. Based on our 25-year experience, the results of 194 patients after a standardized Kausch-Whipple resection for adenocarcinoma of the pancreatic head were analyzed and the prognostic factors were evaluated. Between 1972 and 1998 a total of 221 patients were diagnosed for ductal adenocarcinoma of the pancreatic head, and 194 of them subsequently underwent a standardized Kausch-Whipple resection. Long-term results and prognostic factors were examined by multivariate and univariate analyses. The overall postoperative mortality was 3.09%, and the morbidity was 29.9%. By multivariate analysis only curative resection (R0) was significantly related to a favorable prognosis ( p < 0.0001). Furthermore, in case of a curative resection, the presence of lymph node metastases showed prognostic significance in the multivariate analysis ( p = 0.005). Cumulative survival analysis revealed a 5-year survival rate of 25.4%, a 7-year survival rate of 12.3%, and a 10-year survival rate of 8.2% for patients who underwent curative resection (R0) for adenocarcinoma of the pancreatic head. We demonstrated that the R0 status is the only independent prognostic factor after surgery for adenocarcinoma of the pancreatic head. In the case of a curative resection, the presence of lymph node metastases is of prognostic relevance. In view of considerable surgical morbidity and mortality, resection for cancer of the pancreatic head is the only option if the lesion is resectable. We concluded that surgical treatment is "as good as it gets," as extended techniques have not proved to produce better results.


Subject(s)
Carcinoma, Pancreatic Ductal/surgery , Pancreatic Neoplasms/surgery , Pancreaticoduodenectomy , Carcinoma, Pancreatic Ductal/mortality , Carcinoma, Pancreatic Ductal/pathology , Female , Humans , Male , Multivariate Analysis , Neoplasm Staging , Pancreatic Neoplasms/mortality , Pancreatic Neoplasms/pathology , Pancreaticoduodenectomy/methods , Prognosis , Proportional Hazards Models , Retrospective Studies , Survival Analysis , Treatment Outcome
18.
Am J Surg Pathol ; 26(12): 1578-87, 2002 Dec.
Article in English | MEDLINE | ID: mdl-12459624

ABSTRACT

Cure for ductal adenocarcinoma of the pancreas is restricted to resectable tumors, but survival after surgery is still poor. Despite apparently curative resection, these cancers rapidly recur. Thus, the present pathologic examination should be enriched by sensitive methods to detect minimal residual disease. In a prospective setting we studied the frequency of minimal residual disease after curative resection by routine histopathology, immunohistology, and polymerase chain reaction (PCR) for mutated K-ras. Furthermore, the prognostic implication of detecting of MRD was determined. Prospectively, tumor tissue and corresponding paraaortic lymph nodes were obtained from 78 patients, who underwent surgery for pancreatic head tumors between 1999 and 2001. Sixty-nine of 78 cases were diagnosed for ductal adenocarcinoma (study group), whereas nine cases were diagnosed for benign pancreatic tumors (control group). Paraaortic lymph nodes were examined in step sections by routine histopathology (hematoxylin and eosin) and immunohistology using a pan-cytokeratin antibody. DNA of the primary tumor and corresponding paraaortic lymph nodes were analyzed by PCR-based assays with respect to mutated K-ras in codon 12. The recurrence-free survival and overall survival were correlated with the results of the latter methods. In 3 of 69 patients tumor cells were detected in paraaortic lymph nodes by routine histopathology and in 5 of 69 patients by immunohistology. K-ras mutations were detected in 42 of 69 ductal adenocarcinomas (61%), whereas 12 (17%) were positive in paraaortic lymph nodes. All of the latter patients had recurrence after surgery and a significant poorer survival than those without mutated K-ras. Furthermore, paraaortic lymph nodes diagnosed for K-ras mutation were independent prognostic markers in multivariate analysis. In the control group K-ras mutations were detected in one adenoma of Vater's papilla but not in paraaortic lymph nodes. Tumor cell DNA can be detected more sensitively by the described PCR method than with hematoxylin and eosin or immunohistologic staining, leading to a higher sensitivity for detection of micrometastases. The described PCR method clearly determines subgroups of patients after curative resection with early recurrence and poor survival and could therefore enrich the pathologic examination.


Subject(s)
Adenocarcinoma/genetics , Adenocarcinoma/pathology , Genes, ras , Mutation , Pancreatic Neoplasms/genetics , Pancreatic Neoplasms/pathology , Adenocarcinoma/surgery , Aged , Aorta , Chronic Disease , Cystadenoma/genetics , Cystadenoma/pathology , DNA, Neoplasm/analysis , Diagnostic Tests, Routine , Female , Humans , Immunohistochemistry/methods , Keratins , Lymphatic Metastasis/diagnosis , Lymphatic Metastasis/genetics , Male , Middle Aged , Multivariate Analysis , Neoplasm Staging , Pancreatic Neoplasms/surgery , Pancreaticoduodenectomy , Pancreatitis/genetics , Pancreatitis/pathology , Polymerase Chain Reaction/methods , Polymorphism, Restriction Fragment Length , Predictive Value of Tests , Prognosis , Sequence Analysis, DNA , Survival Analysis
19.
Eur J Surg ; 168(6): 339-44, 2002.
Article in English | MEDLINE | ID: mdl-12428871

ABSTRACT

OBJECTIVE: To find out if resections of cancers of the head of pancreatic are justified in patients over the age of 70 years. DESIGN: Retrospective study. SETTING: University hospital, Germany. SUBJECTS: 519 patients with cancers of the pancreatic head, 93 (18%) of whom were aged 70 or over. MAIN OUTCOME MEASURES: Comparison of outcomes between those aged 70 or over, and those aged less than 70. RESULTS: There were 247 ductal adenocarcinomas, 134 carcinomas of the papilla of Vater, 79 carcinomas of the distal common bile duct, and 59 miscellaneous tumours. Of all variables compared (age, sex, symptoms, operations, clinical and pathological stage. morbidity, mortality, and long-term survival) the only significant difference between the groups was that leaks from the pancreaticojejunostomy occured more often in the older age group (p = 0.02). However, this did not influence overall morbidity or mortality. CONCLUSION: Patients' age is not a limiting factor in attempts at curative resection of cancers of the head of pancreas. If the tumour is resectable and patient is motivated and well enough, resection is indicated whatever the age.


Subject(s)
Pancreatic Neoplasms/surgery , Age Factors , Aged , Aged, 80 and over , Ampulla of Vater , Carcinoma, Pancreatic Ductal/pathology , Carcinoma, Pancreatic Ductal/surgery , Common Bile Duct Neoplasms/mortality , Common Bile Duct Neoplasms/surgery , Female , Humans , Male , Neoplasm Staging , Pancreatic Neoplasms/mortality , Pancreatic Neoplasms/pathology , Patient Selection , Retrospective Studies , Survival Analysis , Treatment Outcome
20.
Pancreatology ; 2(5): 449-55, 2002.
Article in English | MEDLINE | ID: mdl-12378112

ABSTRACT

BACKGROUND: Infections are frequent complications and determine clinical course and outcome in severe pancreatitis. A novel animal model was used to assess minimal transit time of bacterial translocation (BT) across the gut mucosa in vivo using green fluorescent protein-transfected Escherichia coli and intravital video microscopy. METHODS: Three hours after induction of acute pancreatitis by i.p. injection of 40 microg/kg cerulein, 0.5 ml of a suspension of green fluorescent protein-transfected E. coli were injected into the lumen of a small bowel reservoir formed by ligature in anesthetized Wistar rats. Translocation of E. coli was assessed by intravital microscopy. Animals were sacrificed 5 h after induction of pancreatitis. RESULTS: BT across the mucosa and into the muscularis propria took a mean +/- SD of 36.4 +/- 8 min and 80.9 +/- 9.5 min, respectively, in sham animals. Pancreatitis resulted in a significantly shorter minimal transit time across the mucosa (16.4 +/- 4.9 min, p = 0.007) and into the muscularis propria (47.7 +/- 2.5 min, p = 0.001). E. coli were detected on frozen cross-sections and on bacteriological examination of pancreatic tissue in animals with acute pancreatitis but not in controls. DISCUSSION: Intravital microscopy of fluorescent bacteria is a new approach towards studying BT in vivo. Minimal transit time of BT serves as a novel functional aspect of mucosal barrier function during acute pancreatitis. The observation of fluorescent bacteria translocating from the small bowel lumen into the pancreas provides substantial experimental proof for the gut-origin-hypothesis of infectious complications in pancreatitis.


Subject(s)
Escherichia coli/isolation & purification , Ileum/microbiology , Intestinal Mucosa/microbiology , Pancreatitis/microbiology , Acute Disease , Animals , Ceruletide , Disease Models, Animal , Male , Microscopy, Video , Muscle, Smooth/microbiology , Necrosis , Pancreatitis/chemically induced , Pancreatitis/pathology , Rats , Rats, Wistar
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