Jürgen Ordemann

A novel human gastric primary cell culture system for modelling Helicobacter pylori infection in vitro

Background and aims Helicobacter pylori is the causative agent of gastric diseases and the main risk factor in the development of gastric adenocarcinoma. In vitro studies with this bacterial pathogen largely rely on the use of transformed cell lines as infection model. However, this approach is intrinsically artificial and especially inappropriate when it comes to investigating the mechanisms of cancerogenesis. Moreover, common cell lines are often defective in crucial signalling pathways relevant to infection and cancer. A long-lived primary cell system would be preferable in order to better approximate the human in vivo situation. Methods Gastric glands were isolated from healthy human stomach tissue and grown in Matrigel containing media supplemented with various growth factors, developmental regulators and apoptosis inhibitors to generate long-lasting normal epithelial cell cultures. Results Culture conditions were developed which support the formation and quasi-indefinite growth of three dimensional (3D) spheroids derived from various sites of the human stomach. Spheroids could be differentiated to gastric organoids after withdrawal of Wnt3A and R-spondin1 from the medium. The 3D cultures exhibit typical morphological features of human stomach tissue. Transfer of sheared spheroids into 2D culture led to the formation of dense planar cultures of polarised epithelial cells serving as a suitable in vitro model of H. pylori infection. Conclusions A robust and quasi-immortal 3D organoid model has been established, which is considered instrumental for future research aimed to understand the underlying mechanisms of infection, mucosal immunity and cancer of the human stomach.

The COP9 signalosome, cullin 3 and Keap1 supercomplex regulates CHOP stability and adipogenesis.

Summary Obesity is one of the most serious health problems of the 21st century. It is associated with highly increased risk of type 2 diabetes, high blood pressure, cardiovascular disease as well as several cancers. The expansion of the fat tissue needs the differentiation of preadipocytes to adipocytes, a process called adipogenesis. Dysfunction of adipogenesis is a hallmark of obesity and delineation of underlying mechanisms has high priority for identifying targets for pharmacological intervention. Here we investigate the impact of the COP9 signalosome (CSN), a regulator of cullin-RING ubiquitin ligases (CRLs), and of C/EBP homologous protein (CHOP) on the differentiation of LiSa-2 preadipocytes. CHOP induced by piceatannol or by permanent overexpression in LiSa-2 cells blocks adipocyte differentiation as characterized by inhibited fat droplet formation and vascular endothelial growth factor (VEGF) production. Knockdown of the CSN by permanent downregulation of CSN1 in LiSa-2 cells elevates CHOP and retards adipogenesis. The effect of the CSN knockdown on CHOP stability can be explained by the protection of the CRL component Keap1 by the CSN associated ubiquitin-specific protease 15 (USP15). Pulldowns and glycerol gradients reveal that CHOP interacts with a supercomplex consisting of the CSN, cullin 3 and Keap1. Transient knockdown of Keap1 increases CHOP steady state level and retards its degradation. We conclude that CHOP stability is controlled by a CSN-CRL3Keap1 complex, which is crucial for adipogenesis. Our data show that CHOP is a distinguished target for pharmacological intervention of obesity.

Isolated Amyloid Tumor in the Mediastinum: Report of a Case

Abstract.Primary amyloidosis isolated in the mediastinum is rarely encountered in thoracic surgery and few such cases have been reported. We present a case of primary isolated hilar amyloidosis of the mediastinum to illustrate the difficulties in differentiating this disorder preoperatively from central bronchial carcinoma, carcinoid tumor, and mediastinal lymphoma. Usually, a definitive diagnosis can only be made by open biopsy during thoracoscopy or thoracotomy. In conclusion, amyloidosis should be considered in the differential diagnosis of patients when calcifications are found, bearing in mind that radiologic findings are inconclusive and transbronchial biopsy can be negative.

CAND1 exchange factor promotes Keap1 integration into cullin 3-RING ubiquitin ligase during adipogenesis

Adipogenesis is governed by a plethora of regulatory proteins which are most commonly controlled by the ubiquitin proteasome system. Here, we show that the differentiation of LiSa-2 preadipocytes is associated with an increase of cullin-associated and neddylation-dissociated 1 (CAND1), COP9 signalosome (CSN), neddylated cullin 3 (Cul3) and the BTB protein Keap1. Silencing of CAND1 leads to a decrease and reduced integration of Keap1 into Cul3-RING ubiquitin ligases (CRL3) and to a retardation of adipogenesis. Transient transfection of LiSa-2 cells with CAND1 targeting miRNA148a also reduces Keap1 and slowed down adipogenesis of LiSa-2 cells. These results demonstrate for the first time that CAND1 acts as a BTB-protein exchange factor for CRL3 complexes. The specific increase of neddylated Cul3 might be explained by the recruitment of Cul3 or CRL3 in a membrane-bound location during adipogenesis. Together, the results show that during adipogenesis in LiSa-2 cells a CAND1-dependent remodeling and activation/neddylation of CRL3 complexes take place.

Changes in self-reported eating patterns after laparoscopic sleeve gastrectomy: a pre-post analysis and comparison with conservatively treated patients with obesity.

BACKGROUND Patients with severe obesity need to adapt to surgically induced changes in their eating behaviors to maintain treatment success. OBJECTIVES This study examined the effects of laparoscopic sleeve gastrectomy (LSG) on weight loss and on 3 dimensions of eating behavior, namely, cognitive restraint, disinhibition, and hunger. Outcomes of the LSG group were compared with a group of conservatively treated (CT) patients, who underwent a 1-year multimodal weight-reduction group program that included dietary advice, physical exercise, psychoeducation, cognitive-behavioral therapy, training in Jacobsons progressive muscle relaxation, and social group support. SETTING The study setting was a multidisciplinary obesity center located in a university hospital. METHODS A sample of 102 patients with obesity were investigated using the Three-Factor Eating Questionnaire before and, on average, 19 (±5) months after weight loss intervention. Of the 102 patients, 62 (age 45.8±10.8 years, 71% females) underwent LSG, and 40 patients (age 50.6±11.3 years, 77.5% females) underwent the CT program. Patients were assigned to either the surgical or the nonsurgical intervention group following clinical guidelines and patient preference. RESULTS In the LSG group, total weight loss was 25.9±11.0%, excess weight loss was 52.8±24.1%, and body mass index decreased from 51.4±8.1 to 38.0±7.8 kg/m². In the CT group, total weight loss was 5.4±10.6%, excess weight loss was 13.9±27.1%, and body mass index decreased from 40.3±6.7 to 38.0±7.2 kg/m². Significant improvements in self-reported eating behaviors were observed in both groups, that is, an increased cognitive restraint of eating, a decreased disinhibition of eating control, and a reduced degree of perceived hunger. In contrast, whereas Three-Factor Eating Questionnaire scores before weight loss intervention did not differ between groups, LSG patients reported significantly greater reductions in disinhibition and hunger than CT patients did after weight loss intervention. In both groups, greater weight loss was associated with decreased hunger sensations. CONCLUSION In the second follow-up year, LSG was associated with greater weight loss and greater improvements in self-reported eating behaviors compared with conservative treatment.

Verfahrenswahl und Technik der metabolischen Chirurgie

ZusammenfassungDie metabolische Chirurgie entwickelt sich zu einer eindrucksvollen Therapieoption des Diabetes mellitus Typ 2 und anderen metabolischen Erkrankungen. Im Vergleich zur konservativen Therapie können mit bariatrischen Verfahren (Magenbypass, Schlauchmagen, Magenband und biliopankreatischer Diversion) sowohl höhere Remissionsraten als auch eine deutliche Verbesserungen des Blutzuckerstoffwechsels erreicht werden. Zunehmend beschreiben Studien Wirkmechanismen, die über das bisherige Verständnis wie Restriktion und Malabsorption hinausgehen. Die aktuelle Literatur deutet darauf hin, dass Magenbypass und Schlauchmagen das günstigste Nutzen-Risiko-Profil erzielen. Magenband und die biliopankreatische Diversion sind in Einzelfällen zu empfehlen.AbstractMetabolic surgery is becoming an impressive therapeutic option for type 2 diabetes mellitus and other metabolic diseases. Compared to conservative therapy bariatric procedures, such as gastric bypass, sleeve gastrectomy, gastric banding and biliopancreatic diversion, seem to achieve significantly higher remission rates and improvements in blood glucose metabolism. Recent studies describe additional effect mechanisms which go beyond the assumed mechanisms of restriction and malabsorption. The results in the current literature suggest that gastric bypass and sleeve gastrectomy provide the best metabolic risk-benefit profiles. Gastric banding and biliopancreatic diversion can only be recommended in specific cases.Metabolic surgery is becoming an impressive therapeutic option for type 2 diabetes mellitus and other metabolic diseases. Compared to conservative therapy bariatric procedures, such as gastric bypass, sleeve gastrectomy, gastric banding and biliopancreatic diversion, seem to achieve significantly higher remission rates and improvements in blood glucose metabolism. Recent studies describe additional effect mechanisms which go beyond the assumed mechanisms of restriction and malabsorption. The results in the current literature suggest that gastric bypass and sleeve gastrectomy provide the best metabolic risk-benefit profiles. Gastric banding and biliopancreatic diversion can only be recommended in specific cases.

Einfluß verschiedener Gase in der Laparoskopie auf das Tumorzellwachstum. Vorläufige Ergebnisse einer experimentellen Studie im Rattenmodell

Eine potentielle Gefahr laparoskopischer Operationen von malignen Tumoren stellt die Entwicklung von Tumormetastasen in den Trokaren und das fruhzeitige Auftreten von Tumorrezidiven dar. Trotz unklarer Pathogenese, wird hauptsachlich die instrumenteile Manipulation sowie eine hierdurch bedingte Kontamination der Instrumente und der Bauchhohle fur das Auftreten dieser Phanomene verantwortlich gemacht [1,2,3]. Zusatzlich wird die mimeiformige Wundflache der Trokarinzision als optimale Vorraussetzung fur eine Implantation von Tumorzellen angesehen [1]. O’Rourke et al. berichteten uber 3 Rezidive, die bereits 3 Wochen nach laparoskopischer Entfernung eines Gallenblasenkarzinoms auftraten [3]. Die beschriebenen Rezidive traten nicht nur im Bereich der Bergeinzision, sondern auch in anderen Trokarinzisionen auf. Die Implantation von Tumorzellen in die Bauchwand ist deshalb nicht allein durch die lokale Verschleppung von Tumorzellen bei der Entfernung des Tumors zu erklaren. Eine mogliche Ursache des vermehrten Tumorzellwachstums konnte in der Verwendung von Kohlendioxid zum Aufbau des Pneumoperitoneums liegen. Ein vermehrtes Tumorwachstum ware theoretisch durch eine direkte Beeinflusung der Zellen selbst oder aber durch Veranderung der immunologischen Funktion des Patienten denkbar. Impfmetastasen sind in der offenen Chirurgie sehr selten und konnten ein typisches Phanomen der laparoskopischen Chirurgie darstellen, welches diese Technik bei malignen Erkrankungen in Frage stellen wurde. In einer experimentellen Studie haben wir deshalb zunachst invitro den Einflus von Kohlendioxid, sowie Luft und Helium auf das Tumorzellwachstum untersucht und die Ergebnisse im Rattenmodell uberpruft.

Improvement in self-reported eating-related psychopathology and physical health-related quality of life after laparoscopic sleeve gastrectomy: A pre-post analysis and comparison with conservatively treated patients with obesity

OBJECTIVE The present study examined the effects of laparoscopic sleeve gastrectomy (LSG) on self-reported eating-related psychopathology and health-related quality of life (HRQoL). Outcomes of the LSG group were compared with a group of conservatively treated (CT) patients, who underwent a 1-year multimodal weight reduction group program that included dietary advice, physical exercise, psychoeducation, cognitive-behavioral therapy, training in Jacobsons progressive muscle relaxation, and social group support. The setting was a multidisciplinary obesity center. METHOD A sample of 103 patients with obesity were investigated using the Eating Disorder Inventory and the Short Form Health Survey before and, on average, 19 (±5) months after weight loss intervention. Thereof, 63 patients (age 45.6±10.9years, 71.4% females) underwent LSG, and 40 patients (age 50.6±11.3years, 77.5% females) underwent the CT program. Patients were assigned to either the surgical or the nonsurgical intervention group following clinical guidelines and patient preference. RESULTS In the LSG group, excess weight loss (%EWL) was 53.0±24.0%, and body mass index (BMI) decreased from 51.5±8.1 to 38.0±7.7kg/m2. In the CT group, %EWL was 13.9±27.1%, and BMI decreased from 40.3±6.7 to 38.0±7.2kg/m2. Significant improvements in eating-related psychopathology were observed in both groups. Although both groups had a similar BMI after the respective interventions, LSG patients reported significantly greater body satisfaction and substantial improvement in perceived physical health from a lower baseline level than CT patients. DISCUSSION In the second follow-up year, LSG was associated with greater weight loss from a higher baseline weight, and greater improvements in self-reported eating-related psychopathology and physical HRQoL compared with conservative treatment.

Verfahrenswahl und Technik der metabolischen Chirurgie@@@Procedure selection and technique of metabolic surgery

ZusammenfassungDie metabolische Chirurgie entwickelt sich zu einer eindrucksvollen Therapieoption des Diabetes mellitus Typ 2 und anderen metabolischen Erkrankungen. Im Vergleich zur konservativen Therapie können mit bariatrischen Verfahren (Magenbypass, Schlauchmagen, Magenband und biliopankreatischer Diversion) sowohl höhere Remissionsraten als auch eine deutliche Verbesserungen des Blutzuckerstoffwechsels erreicht werden. Zunehmend beschreiben Studien Wirkmechanismen, die über das bisherige Verständnis wie Restriktion und Malabsorption hinausgehen. Die aktuelle Literatur deutet darauf hin, dass Magenbypass und Schlauchmagen das günstigste Nutzen-Risiko-Profil erzielen. Magenband und die biliopankreatische Diversion sind in Einzelfällen zu empfehlen.AbstractMetabolic surgery is becoming an impressive therapeutic option for type 2 diabetes mellitus and other metabolic diseases. Compared to conservative therapy bariatric procedures, such as gastric bypass, sleeve gastrectomy, gastric banding and biliopancreatic diversion, seem to achieve significantly higher remission rates and improvements in blood glucose metabolism. Recent studies describe additional effect mechanisms which go beyond the assumed mechanisms of restriction and malabsorption. The results in the current literature suggest that gastric bypass and sleeve gastrectomy provide the best metabolic risk-benefit profiles. Gastric banding and biliopancreatic diversion can only be recommended in specific cases.Metabolic surgery is becoming an impressive therapeutic option for type 2 diabetes mellitus and other metabolic diseases. Compared to conservative therapy bariatric procedures, such as gastric bypass, sleeve gastrectomy, gastric banding and biliopancreatic diversion, seem to achieve significantly higher remission rates and improvements in blood glucose metabolism. Recent studies describe additional effect mechanisms which go beyond the assumed mechanisms of restriction and malabsorption. The results in the current literature suggest that gastric bypass and sleeve gastrectomy provide the best metabolic risk-benefit profiles. Gastric banding and biliopancreatic diversion can only be recommended in specific cases.

Procedure selection and technique of metabolic surgery

ZusammenfassungDie metabolische Chirurgie entwickelt sich zu einer eindrucksvollen Therapieoption des Diabetes mellitus Typ 2 und anderen metabolischen Erkrankungen. Im Vergleich zur konservativen Therapie können mit bariatrischen Verfahren (Magenbypass, Schlauchmagen, Magenband und biliopankreatischer Diversion) sowohl höhere Remissionsraten als auch eine deutliche Verbesserungen des Blutzuckerstoffwechsels erreicht werden. Zunehmend beschreiben Studien Wirkmechanismen, die über das bisherige Verständnis wie Restriktion und Malabsorption hinausgehen. Die aktuelle Literatur deutet darauf hin, dass Magenbypass und Schlauchmagen das günstigste Nutzen-Risiko-Profil erzielen. Magenband und die biliopankreatische Diversion sind in Einzelfällen zu empfehlen.AbstractMetabolic surgery is becoming an impressive therapeutic option for type 2 diabetes mellitus and other metabolic diseases. Compared to conservative therapy bariatric procedures, such as gastric bypass, sleeve gastrectomy, gastric banding and biliopancreatic diversion, seem to achieve significantly higher remission rates and improvements in blood glucose metabolism. Recent studies describe additional effect mechanisms which go beyond the assumed mechanisms of restriction and malabsorption. The results in the current literature suggest that gastric bypass and sleeve gastrectomy provide the best metabolic risk-benefit profiles. Gastric banding and biliopancreatic diversion can only be recommended in specific cases.Metabolic surgery is becoming an impressive therapeutic option for type 2 diabetes mellitus and other metabolic diseases. Compared to conservative therapy bariatric procedures, such as gastric bypass, sleeve gastrectomy, gastric banding and biliopancreatic diversion, seem to achieve significantly higher remission rates and improvements in blood glucose metabolism. Recent studies describe additional effect mechanisms which go beyond the assumed mechanisms of restriction and malabsorption. The results in the current literature suggest that gastric bypass and sleeve gastrectomy provide the best metabolic risk-benefit profiles. Gastric banding and biliopancreatic diversion can only be recommended in specific cases.