Downregulation of the ubiquitin-proteasome system in normal colonic macrophages and reinduction in inflammatory bowel disease.

BACKGROUND: In normal mucosa, intestinal lamina propria macrophages (IMACs) maintain tolerance against food antigens and the commensal bacterial flora. Several mechanisms have been identified that mediate tolerance. The ubiquitin-proteasome system (UPS) is a large multiprotein complex that degrades cellular proteins. As the UPS may modulate immune functions of IMACs, we performed a detailed investigation of UPS expression and function under normal conditions and in cells derived from patients suffering from inflammatory bowel disease (IBD). METHODS: IMACs were isolated from intestinal mucosa. mRNA expression of macrophages differentiated in vitro (i.v. MACs) and IMACs was compared by Affymetrix® oligonucleotide arrays. Quantitative Taqman-PCR was performed on five exemplary proteasomal and five ubiquitinylation genes each. Proteins were analyzed by immunohistochemistry and Western blotting. Proteasome function was assessed by a fluorimetric test. RESULTS: Affymetrix analysis showed downregulation of mRNA expression of almost all represented proteasomal and of 22 ubiquitination-associated genes in IMACs as compared to i.v. MACs and monocytes. By quantitative PCR, up to tenfold higher mRNA expression of 10 exemplary genes of the UPS (UBE2A, UBE2D2, UBE2L6, USP14, UBB and ATPase2, ß2, ß5, ß2i/MECL-1, ß5i/LMP7) was demonstrated in i.v. MACs as compared to IMACs. Immunohistochemistry and Western blots confirmed these findings in intestinal mucosa of controls and patients suffering from diverticulitis. In contrast, a significant increase in protein amounts was found in mucosa of patients with IBD. CONCLUSION: Reduced expression of subunits of the UPS in IMACs of normal mucosa supports the concept of the presence of a nonreactive, anergic macrophage phenotype in the gut under normal conditions. Reinduction in IMACs of IBD mucosa reflects activated IMACs which can present antigenic peptides and thus support inflammation.

Multidetector computed tomography mesentericography for the diagnosis of obscure gastrointestinal bleeding.

PURPOSE: To evaluate the diagnostic yield of 16-row multidetector computed tomography (CT) mesentericography in patients with obscure gastrointestinal bleeding. MATERIALS AND METHODS: The radiological information system database was used to retrospectively identify all patients in whom CT mesentericography (CTM) was performed for the diagnosis of obscure gastrointestinal bleeding between July 2002 and September 2006. A subsequent prospective study was conducted between October 2006 and September 2009 to evaluate CTM in patients with major obscure gastrointestinal bleeding. The retrospectively identified patients (six patients) as well as the prospectively evaluated patients (seven patients) constitute the study population. Following mesenteric DSA the catheter was left in the superior mesenteric artery, the patient was transferred to the CT suite and CTM was carried out by scanning the abdomen after contrast material injection via the catheter. Active bleeding was suspected if a focal area of high attenuation consistent with contrast material extravasation was found within the bowel lumen. RESULTS: CTM detected the site of active bleeding in three of 13 patients (23 %). In the subpopulation of patients who were prospectively evaluated, CT mesentericography identified the site of active bleeding in one of seven patients (14 %). Depiction of active bleeding by CTM prompted surgical intervention in each case and surgery confirmed the findings of CT mesentericography. CONCLUSION: Due to the relatively low rate of positive findings and inherent drawbacks, we feel that CTM cannot be recommended in general. However, in selected patients who are continuously bleeding at a low rate and in whom iv-CT was negative, CT mesentericography might be helpful.

Giant cell hepatitis: an unusual cause of fulminant liver failure.

Giant cell hepatitis is a very rare disease of unknown origin. It has been hypothesized that drugs, viral infections, or autoimmune reactions may play a pathogenetic role. Here, we describe a 33 year old patient with bacterial bronchitis who was treated with doxycycline (100 mg/d) for one week. Furthermore the patient complained of malaise and a distinct jaundice. Liver parameters increased dramatically (AST 4670 U/l, ALT 5350 U/l, bilirubin 226 µmol/l) and liver function was impaired (INR = 1,45). The ultrasound scan showed a hepatomegaly with no signs of cirrhosis, normal spleen size and normal bile ducts; liver perfusion was normal. No evidence of Wilson's disease, hemochromatosis, autoimmune hepatitis, primary sclerosing cholangitis, primary biliary cirrhosis, hepatitis A, B, C and E, HIV, CMV, VZV, adenoviral infections, or paracetamol intoxication was found. Subsequently, the patient developed acute liver failure (AST 2134 U/l, ALT 2820 U/l, bilirubin 380 µmol/l, INR 3.0) and a beginning renal failure. Therefore, he was transferred to our transplant center. Due to increasing confusion and somnolence due to cerebral edema mechanical ventilation was needed. Because of an acute renal failure and severe hepatic encephalopathia MARS-hemodialysis was performed. Three weeks after the appearance of the jaundice he underwent liver transplantation (MELD 40). Surprisingly, in the explanted liver the diagnosis of giant cell hepatitis was made. Today--2 years after successful liver transplantation--the patient is in very good condition with normal liver function. In conclusion, giant cell hepatitis is a rare cause of acute liver failure that is often recognized only histologically.

Sono-hepatic-arteriography (Sono-HA) in the assessment of hepatocellular carcinoma in patients undergoing transcatheter arterial chemoembolization (TACE).

PURPOSE: The aim of our study was to evaluate sono-hepatic-arteriography in patients with hepatocellular carcinoma undergoing transcatheter arterial chemoembolization. MATERIALS AND METHODS: We evaluated 15 patients with hepatocellular carcinoma undergoing TACE who presented in our institution from February 2006 to May 2008. All patients underwent a conventional B-mode ultrasound examination using a high-end machine and a multi-frequency transducer (2.5 - 4 MHz) before dynamic contrast-enhanced ultrasound examination was carried out. For the sono-hepatic-arteriography 1 ml SonoVue was injected as a bolus using the formerly placed intraarterial catheter. Biphasic enhanced computed tomography was performed using a 16-slice CT scanner up to 48 hours before transcatheter arterial chemoembolization and during follow-up. RESULTS: The lesion size (of the largest lesion) ranged from 1 to 13 centimeters in their largest diameter (mean: 4.8 cm). Contrast-enhanced ultrasound diagnosed more lesions than B-mode sonography in eight cases and more lesions than computed tomography in 5 patients. The findings of sono-hepatic-arteriography were correct in fourteen cases (93.3 %). Direct impact on patient management was seen in eleven patients (73.3 %). CONCLUSION: We were able to show that the application of an intraarterial sonographic contrast agent during embolization is able to diagnose new lesions on the one hand and to assess the embolization success on the other. This might improve transcatheter arterial chemoembolization results and patient outcome.

Decreased cytosolic glucocorticoid receptor levels in critically ill patients.

The immune system and the hypothalamic-pituitary-adrenal axis are linked by several mechanisms. Intracellular glucocorticoid receptors represent one important connection. The aim of this study was to examine the coherence between the number of glucocorticoid receptors, activation of the hypothalamic-pituitary-adrenal axis, inflammatory cytokine levels and the severity of illness in critically ill patients. In a prospective study, blood was collected from 20 healthy members of the hospital staff (control group) and 50 ventilated patients (sample group) within the first 24 hours after intubation and within three days of extubation. 3H-dexamethasone-binding assay was used to assess cytoplasmatic free glucocorticoid receptors levels. ACTH, cortisol, IL-6 and TNFa levels were measured by ELISA. In the sample group, specific binding of 3H-dexamethasone was significantly decreased compared to the control group. Glucocorticoid receptor levels tended to be lower in more severely ill patients. Plasma cortisol and ACTH levels were significantly different from the control group after extubation but not at intubation. Severe illness is associated with rapid down-regulation of 3H-dexamethasone binding. This decrease occurs before elevation of plasma cortisol. Therefore, down-regulation of cortisol binding may be directly associated with the stress response and not due to feedback regulation following increase in plasma cortisol levels.

[Localisation of hepatic lesions to liver segments according to sonography (US), computed tomography (CT) and surgery (OP): a comparative study]. / Untersuchung der Segmentzugehörigkeit von Leberraumforderungen in Sonografie (US), Computertomografie (CT) und intraoperativ (OP): eine Vergleichsstudie.

INTRODUCTION: The localisation of focal liver lesions is usually performed according to the Couinaud classification system. The exact description of localisation and size of liver lesions is especially important for surgical procedures. The aim of this prospective study was the evaluation of differences and agreements in the localisation and size of hepatic lesions as found by ultrasound (US), computed tomography (CT) and according to the intraoperative status (OP). MATERIAL AND METHODS: 32 patients (21 male, 11 female) were enrolled in the study. The results obtained from sonography, computed tomography and surgery were classified into 5 categories for localisation and for size, respectively. RESULTS: According to the agreement between sonography and computed tomography, 25 % of all hepatic lesions were classified into category 1 (exact agreement), whereas 40.6 % were ranked into category 2 (almost exact agreement). Correlating sonography and intraoperative results, 31.3 % of the lesions were classified into category 1 and 46.9 % into category 2. In the comparison of CT with OP, 34.4 % of the lesions were found to be in category 1 and 43.8 % in category 2. Concerning the size of the lesions, almost half of the tumours (46.9 %) were classified into category 1 on the basis of the correlation between US and CT and 21.9 % on the basis of the correlation between US and OP. DISCUSSION: The localisation and description of the size of hepatic lesions is mainly similar or even identical on the basis of the different methods. Further improvements might be achieved by the introduction of a consistent nomenclature.

Severe acute pancreatitis requiring drainage therapy: findings on computed tomography as predictor of patient outcome.

BACKGROUND/AIMS: To evaluate whether morphologic features on computed tomography (CT) correlate with outcome of patients with severe acute pancreatitis (SAP). METHODS: 80 patients with SAP requiring percutaneous drainage therapy were retrospectively analyzed. Twelve CT features beyond the CT severity index (CTSI) were studied. Endpoints for patient outcome were patient death, length of hospital and ICU stay. The twelve features and the CTSI score were correlated with mortality using Kaplan-Meier estimator and correlated with length of hospital and ICU stay using the χ(2) test. A p value ≤0.05 was considered statistically significant. RESULTS: Two CT features exhibited a significant correlation with mortality: (1) the number of parts of pancreas (head, corpus, tail) that exhibited areas of necrosis and (2) the presence of distant fluid collections (posterior pararenal space and/or paracolic gutter). Mortality was 42% (21 of 50 patients) and 20% (6 of 30 patients) if two/all three parts or none/one part of the pancreas exhibited necrosis, respectively. Mortality was 46% (18 of 39 patients) and 22% (9 of 41 patients) if distant fluid collections were present or absent, respectively. All other imaging features including the CTSI showed no significant correlation with patient outcome. CONCLUSION: We identified two morphologic features on CT that might be helpful to predict prognosis of patients suffering from SAP. and IAP.

Recipient NOD2/CARD15 status affects cellular infiltrates in human intestinal graft-versus-host disease.

Nucleotide-binding oligomerization domain 2/caspase recruitment domain 15 (NOD2/CARD15) polymorphisms have been identified as risk factors of both Crohn's disease and graft-versus-host disease (GVHD) following allogeneic stem cell transplantation. However, the role of these receptors of innate immunity in the pathophysiology of gastrointestinal GVHD is still poorly defined. Immunohistological features of intestinal GVHD were analysed in gastrointestinal biopsies from 58 patients obtained at the time of first onset of intestinal symptoms. The observed changes were correlated with concomitant risk factors and the presence of polymorphisms within the pathogen recognition receptor gene NOD2/CARD15. Intestinal GVHD was associated with a stage-dependent decrease in CD4 T cell infiltrates and an increase in CD8 T cells in the lamina propria; CD8 infiltrates correlated with extent of apoptosis and consecutive epithelial proliferation. The presence of NOD2/CARD15 variants in the recipient was associated with a significant loss of CD4 T cells: in a semiquantitative analysis, the median CD4 score for patients with wild-type NOD2/CARD15 was 1.1 (range 3), but only 0.4 (range 2) for patients with variants (P = 0.002). This observation was independent from severity of GVHD in multivariate analyses and could not be explained by the loss of forkhead box P3(+) T cells. Our results suggest a loss of protective CD4 T cells in intestinal GVHD which is enhanced further by the presence of NOD2/CARD15 variants. Our study might help to identify more selective therapeutic strategies in the future.

Peritoneal fluid adipokines: ready for prime time?

BACKGROUND: Visceral adipose tissues secret a variety of adipokines; however, it is not known whether they are present in the peritoneal fluid. It was the aim of this study to investigate peritoneal fluid concentrations of novel (cartonectin, omentin) and classical adipokines (leptin, adiponectin, resistin, visfatin) in patients with ascites. MATERIAL AND METHODS: Ninety-six patients (71 men and 25 women) undergoing paracentesis were included. Of these, 76 suffered from liver cirrhosis. Adipokines were measured by enzyme-linked immunosorbent assay or Western blot. RESULTS: Each adipokine was detected in ascites with a broad range. Serum-ascites ratios (SAR) correlated with clinical and laboratory parameters. The main variables influencing peritoneal fluid adipokine concentrations were body mass index (BMI), local inflammation, systemic inflammation and serum adipokine concentrations. Resistin was significantly higher in patients with peritonitis and showed a positive correlation with peripheral leucocytes (white blood cell count). Leptin was correlated with the underlying disease. Visfatin correlated with peripheral white blood cell and C-reactive protein levels. Omentin expression was correlated with ascitic leucocyte count, ascitic albumin concentration and low albumin SAR. BMI was correlated positively with ascitic leptin levels and cartonectin protein levels. CONCLUSIONS: Peritoneal fluid adipokine concentrations are characterized by individual SARs, depend on the presence of peritonitis, and correlate with underlying disease, BMI and systemic inflammation. The data open a new field of research on the role of the peritoneum and visceral adipokines in gastrointestinal diseases.

Nutritional composition in different training stages in young female athletes (swimming) and association with leptin, IGF-1 and estradiol.

OBJECTIVES: To investigate the interaction of serum leptin, IGF-1, estradiol and cortisol in salvia as well as IL-6 with nutritional composition in female athletes (swimming) according to the training protocol (competition (C), sprint (S), recreation (R), endurance (E)). DESIGN/METHODS: In 23 young (10-19 years old) female athletes (Bavarian swimming competition) in different training stages nutritional protocols were evaluated using standardized questionnaires. Body composition was measured by using analysis of bioimpedance. Estradiol, IGF-1, leptin and IL-6 in serum were measured by ELISA. To obtain circadian profiles of cortisol salvia probes were sampled at 4 h intervals and cortisol in saliva was measured. RESULTS: Daily intake of kilocalories varied significantly and was highest during R with the highest percentage of nutritional fat intake (37.3%) when compared to C (28.1%, p=0.001). BMI was associated with leptin in all training stages and IL-6 in R, C and E. Leptin, IGF-1 and cortisol in salvia were dependent on training stages while serum levels of estradiol were not. Nutritional fat intake (p=0.07 in R) as well as serum levels of IGF-1 (p=0.014 in E) were significantly associated with estradiol but not with serum leptin levels or salivary cortisol. CONCLUSIONS: In female athletes nutritional composition has an impact on serum hormones (leptin, IGF-1 as well as estradiol) and may be also on cytokines (IL-6). Leptin, IGF-1 and salivary cortisol levels represent the intensity of physical training and possibly overtraining. In our female athletes no findings indicative of female athlete triad could be demonstrated.

Endoscopic retrograde cholangiography of a hepaticojejunostomy using double balloon enteroscopy.

Examination of a biliodigestive anastomosis presents a diagnostic and therapeutic challenge. Visualization of biliodigestive anastomosis and endoscopic retrograde cholangiography with intervention is possible with a double balloon enteroscope.

[Minimal invasive therapy in a patient with necrotizing pancreatitis and disrupted duct by percutaneous endoscopy: report of a case and review of literature]. / Minimalinvasive Therapie einer Patientin mit nekrotisierender Pankreatitis und unterbrochenem Pankreasgang: Darstellung des Falles und Review der Literatur.

A 68-year-old lady was admitted to a primary care centre with abdominal pain. She presented with a tender abdomen and a lipase level of 29 456 U/L. Cholecystectomy had been performed in 1971. Because her clinical state worsened the patient was transferred to our intensive care unit. Computed tomography and contrast ultrasound revealed necrotising pancreatitis. Antibiotics were started immediately. 10 days later the patient became febrile and an area of liquid necrosis between the spleen and the left kidney was drained under sonographic guidance by a 12 Fr pigtail. Every ten days the drain was replaced by a larger one. ERCP revealed a communication between the pancreatic duct and the growing necrotic cavity. A 7 Fr/ 7 cm plastic stent was put in. After 30 days an unsuccessful endoscopic necrosectomy was attempted with the cholangioscope through a 28 Fr peel-away sheath. In a further session an effective percutaneous necrosectomy was possible through a laparascopy trocar with a standard gastroscope. The patient became afebrile and signs of infection decreased. The persisting communication between the pancreatic duct and the necrotic cavity was closed with onyx. In the following days the dimensions of the necrotic cavity decreased rapidly. Finally the patient could be discharged home in good health. Antibiotics were stopped and the percutaneous drain was taken out after some weeks of ambulatory care.

Percutaneous necrosectomy in patients with acute, necrotizing pancreatitis.

The objective of this retrospective study was to evaluate the outcome of patients with acute necrotizing pancreatitis treated by active percutaneous necrosectomy. By searching the radiological, surgical and internal medicine databases, all patients with acute necrotizing pancreatitis treated by active percutaneous necrosectomy between 1992 and 2004 were identified. Demographic, laboratory, and clinical data, and details about invasive procedures were collected by reviewing patient charts, radiological and surgical reports. The computed tomography severity index (CTSI) scores were determined by reviewing CT images. Eighteen patients were identified. Median Ranson score on admission was 2. The Acute Physiology and Chronic Health Evaluation (APACHE) II score was median 22. Median CTSI score was 7. Initially all patients were treated with CT-guided drainage placement. Because passive drainage proved not to be effective, subsequent minimally invasive, percutaneous necrosectomy was performed. Eight out of 18 patients recovered fully without the need for surgery. Ten of 18 patients required additional surgical necrosectomy. For one of ten patients, percutaneous necrosectomy allowed postponing surgery by 39 days. Four of ten surgically treated patients died: three from septic multiorgan failure, one from pulmonary embolism. Percutaneous minimally invasive necrosectomy can be regarded as a safe and effective complementary treatment modality in patients with necrotizing pancreatitis. It is suitable for a subset of patients to avoid or delay surgery.

Outcome of patients with acute, necrotizing pancreatitis requiring drainage-does drainage size matter?

AIM: To assess the outcome of patients with acute necrotizing pancreatitis treated by percutaneous drainage with special focus on the influence of drainage size and number. METHODS: We performed a retrospective analysis of 80 patients with acute pancreatitis requiring percutaneous drainage therapy for infected necroses. Endpoints were mortality and length of hospital stay. The influence of drainage characteristics such as the median drainage size, the largest drainage size per patient and the total drainage plane per patient on patient outcome was evaluated. RESULTS: Total hospital survival was 66%. Thirty-four patients out of all 80 patients (43%) survived acute necrotizing pancreatitis with percutaneous drainage therapy only. Eighteen patients out of all 80 patients needed additional percutaneous necrosectomy (23%). Ten out of these patients required surgical necrosectomy in addition, 6 patients received open necrosectomy without prior percutaneous necrosectomy. Elective surgery was performed in 3 patients receiving cholecystectomy and one patient receiving resection of the parathyroid gland. The number of drainages ranged from one to fourteen per patient. The drainage diameter ranged from 8 French catheters to 24 French catheters. The median drainage size as well as the largest drainage size used per patient and the total drainage area used per patient did not show statistically significant influence on mortality. CONCLUSION: Percutaneous drainage therapy is an effective tool for treatment of necrotizing pancreatitis. Large bore drainages did not prove to be more effective in controlling the septic focus.

[The diagnostic approach to hepatocellular carcinoma]. / Diagnostik des hepatozellulären Karzinoms.

Risk factors and symptoms of hepatocellular carcinoma (HCC): The main risk factors of HCC include infection with hepatitis B or C virus, as well as alcohol consumption. There are no specific symptoms of HCC, making early diagnosis and detection of the disease difficult. When HCC presents with specific clinical symptoms, the tumour is typically very far advanced. Surveillance in liver cirrhosis: The most common serological marker used in HCC diagnosis is alpha-fetoprotein (AFP), but other tumour markers such as the des-gamma-carboxyprothrombin (DGCP) or fractions of AFP (AFP-L3) exist and there use is discussed in this context. Surveillance should be done by sonography at 6 (to 12) months intervals. The single nodule in the cirrhotic liver: Ultrasound is the most commonly used imaging modality for detecting HCC tumour nodules with a large range of reported sensitivities. HCC may appear as a hypoechoic, isoechoic, or hyperechoic round or oval lesion with intratumoural flow signals on Doppler or power Doppler sonography. The differentiation of smaller malignant lesions in cirrhotic livers can be improved by contrast-enhanced ultrasound (CEUS). Spiral computed tomography (CT) and magnetic resonance imaging (MRI) with and without contrast enhancement play an important role in the diagnosis and staging of HCC. If the vascular pattern on imaging is not typical, biopsy becomes necessary. The patient with known HCC: Different tumour markers are used in the evaluation of tumour progression, prediction of patient outcome and treatment efficacy. Among the various staging systems used in the context of HCC, the Barcelona-Clinic-Liver-Cancer (BCLC) staging system is currently the only staging system that takes into account tumour stage, liver function, physical status and cancer-related symptoms. Beside surgical resection, non-surgical treatments such as percutaneous ethanol injection (PEI), radiofrequency thermoablation (RFTA) and trans-arterial chemoembolisation (TACE) are used. Successful tumour "bridging" with ablative therapy methods can be achieved in carefully selected patients on the waiting list for orthotopic liver transplantation. Contrast-enhanced sonography is able to control the ablation treatment of HCC.

Outcome of hemato-oncologic patients with and without stem cell transplantation in a medical ICU.

OBJECTIVE: To assess the outcome of a mixed population of critical ill patients with haematological malignancies with special focus on the comparison of patients who underwent stem cell transplantation with non-transplanted patients. METHODS: Retrospective, unicentric analysis of 94 critical ill cancer patients in a tertiary care centre in a period of two years time. RESULTS: We analysed different variables at admission as well as different treatment modalities during the ICU stay. We compared the outcome by using chi-square test by Pearson for categorical variables and Kaplan-Meier as well as Cox-Regression for survival analysis. The general patients characteristics did not significantly differ between transplanted and non-transplanted patients. The overall ICU and hospital mortality were 43% and 54%. Considering just patients with mechanical ventilation we found ICU and hospital mortalities of 65% and 82% in the stem cell transplantation group vs. 67% and 74% in the non-transplanted group, respectively. As risk factors for overall mortality in multivariate analysis only the Simplified Acute Physiology Scale II and the need of ventilation remained significant. Between the underlying diseases mortality did not show significant differences at all. CONCLUSIONS: The outcome and prognosis of critical ill cancer patients has generally slightly improved over the last years. Our data show no statistically significant differences regarding outcome and prognosis between stem cell transplanted and non-transplanted patients receiving ICU treatment. A stem cell transplantation should not be considered a strong contraindication for ICU treatment or artificial ventilation.

In vivo treatment with the herbal phenylethanoid acteoside ameliorates intestinal inflammation in dextran sulphate sodium-induced colitis.

Recently we demonstrated that in inflammatory bowel disease (IBD) macrophage-oxidative burst activity is increased and NADPH oxidase mRNA is induced. The herbal phenylethanoid acteoside isolated from Plantago lanceolata L. was shown to exhibit anti-oxidative potential. Using the dextran sulphate sodium (DSS)-induced colitis model, in this study we have assessed whether systemic application of acteoside affects colitis. Colitis was induced by DSS in Balb/c mice. Treatment with acteoside (120, 600 microg/mouse/day) was performed intraperitoneally. The colon lengths were determined. Colonic tissue was scored histologically (max. score 8) by a blinded investigator. T cells isolated from mesenteric lymph nodes (MLN) were stimulated with anti-CD3 antibody in the presence of interleukin (IL)-2 (final concentration 10 U/ml). After incubation for 24 h, IL-1beta, IL-6, IL-12 tumour necrosis factor (TNF)-alpha and interferon (IFN)-gamma levels in supernatants were analysed by the beadlyte cytokine detection system. Histological scoring of colonic tissue revealed that application of acteoside was followed by a significantly improved histological score. In acute colitis the histological score was 3.2 with acteoside versus 5.2 with phosphate-buffered saline (PBS) (P < 0.02). In chronic colitis both 120 microg (3.3 versus 5.2) or 600 microg acteoside (3.0 versus 5.2) significantly ameliorated colitis (both P < 0.02). Stimulated MLN from mice with chronic DSS-induced colitis treated with acteoside showed a significant down-regulation of IFN-gamma secretion (195 pg/ml with 600 microg acteoside versus 612 pg/ml with PBS, P < 0.02). Inhibition of oxidative burst activity with acteoside reduced mucosal tissue damage in DSS colitis and could be a therapeutic alternative for IBD treatment. Further studies of this agent are warranted.

Soluble galectin-3 is a strong, colonic epithelial-cell-derived, lamina propria fibroblast-stimulating factor.

BACKGROUND: Colonic lamina propria fibroblasts (CLPFs) play an important role in the pathogenesis of fibrosis and strictures in Crohn's disease. AIM: To identify colonic epithelial cell (CEC)-derived factors that activate CLPFs. METHODS: Primary human CECs and CLPFs were isolated from control mucosa and interleukin 8 (IL8) of CLPF cultures was quantified by ELISA. Activation of nuclear factor kappaB (NF-kappaB) was shown, and translocation of NF-kappaB was inhibited by a dominant-negative IkappaB-expressing adenovirus. The major CLPF-activating and IL8 inducing protein was purified using fast-performance liquid chromatography (HiPrep 16/60 Sephacryl S-200 High Resolution Column) and sodium dodecyl sulphate gel electrophoresis. RESULTS: A considerable increase in IL8 secretion by CLPFs cultured in CEC-conditioned media compared with that in unconditioned media (155.00 (10.00) pg/microg v 1.434 (0.695) pg/microg) was found. The effect of CEC-conditioned media on CLPF IL8 secretion was NF-kappaB dependent. A protein or DNA array confirmed the involvement of NF-kappaB and activator protein-1. Purification of a candidate band isolated with the use of sodium dodecyl sulphate-polyacrylamide gel electrophoresis and subsequent sequencing showed soluble galectin-3 to be a strong CLPF-activating factor. Depletion of galectin-3 from conditioned media by immunoprecipitation abolished the CLPF stimulatory effect. CONCLUSIONS: Using a classical biochemical approach, soluble galectin-3 was identified as a strong activator of CLPFs produced by CEC. Galectin-3 induced NF-kappaB activation and IL8 secretion in these cells and may be a target for future therapeutic approaches to reduce or avoid stricture formation.

[Surgical management of patients with peritoneal carcinomatosis of gastrointestinal origin]. / Chirurgische behandlung von peritoneal disseminierten gastrointestinalen tumoren.

Peritoneal carcinomatosis is found in approximately 15 % of patients with colorectal cancer during the course of their disease, and is associated with a poor prognosis. Even more patients with gastric cancer develop peritoneal seeding. Cytoreductive surgery and hyperthermic intraperitoneal chemotherapy (HIPEC) have been introduced in the past decade and have led to 5-year survival rates of 30 to 40 % for selected patients with colorectal cancer and peritoneal carcinomatosis. These numbers have been demonstrated by many retrospective analyses and by prospective Phase II studies. The clinical assessment to select patients who will benefit from the combined therapy and achievement of complete macroscopic cytoreduction both play a crucial role. Less favourabale results have been achieved for patients suffering from stage IV gastric cancer with peritoneal seeding. Promising results were demonstrated for postoperative intraperitoneal chemotherapy following curative gastrectomy. Patients with hepatic, biliary and pancreatic cancers and peritoneal carcinomatosis do not benefit from cytoreductive surgery. There is a need for further multicentre, prospective trials analysing the use of hyperthermic intraperitoneal chemotherapy. They should be conducted in the specialised centres by interdisciplinary teams.

Monocyte chemoattractant protein-1 (MCP-1) inhibits the intestinal-like differentiation of monocytes.

Monocytes (MO) migrating into normal, non-inflamed intestinal mucosa undergo a specific differentiation resulting in a non-reactive, tolerogenic intestinal macrophage (IMAC). Recently we demonstrated the differentiation of MO into an intestinal-like macrophage (MAC) phenotype in vitro in a three-dimensional cell culture model (multi-cellular spheroid or MCS model). In the mucosa of patients with inflammatory bowel disease (IBD) in addition to normal IMAC, a reactive MAC population as well as increased levels of monocyte chemoattractant protein 1 (MCP-1) is found. The aim of this study was to investigate the influence of MCP-1 on the differentiation of MO into IMAC. MCS were generated from adenovirally transfected HT-29 cells overexpressing MCP-1, macrophage inflammatory protein 3 alpha (MIP-3alpha) or non-transfected controls and co-cultured with freshly elutriated blood MO. After 7 days of co-culture MCS were harvested, and expression of the surface antigens CD33 and CD14 as well as the intracellular MAC marker CD68 was determined by flow-cytometry or immunohistochemistry. MCP-1 and MIP-3alpha expression by HT-29 cells in the MCS was increased by transfection at the time of MCS formation. In contrast to MIP-3alpha, MCP-1 overexpression induced a massive migration of MO into the three-dimensional aggregates. Differentiation of IMAC was disturbed in MCP-1-transfected MCS compared to experiments with non-transfected control aggregates, or the MIP-3alpha-transfected MCS, as indicated by high CD14 expression of MO/IMAC cultured inside the MCP-1-transfected MCS, as shown by immunohistochemistry and FACS analysis. Neutralization of MCP-1 was followed by an almost complete absence of monocyte migration into the MCS. MCP-1 induced migration of MO into three-dimensional spheroids generated from HT-29 cells and inhibited intestinal-like differentiation of blood MO into IMAC. It may be speculated that MCP-1 could play a role in the disturbed IMAC differentiation in IBD mucosa.