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Jólkowska, Justyna ; Pieczonka, Anna ; Strabel, Tomasz ; Boruczkowski, Dariusz ; Wachowiak, Jacek ; Bader, Peter ; Witt, Michal
Background: Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is performed mainly in patients with high-risk or advanced hematologic malignancies and congenital or acquired aplastic anemias. In the context of the significant risk of graft failure after allo-HSCT from alternative donors and the risk of relapse in recipients transplanted for malignancy, the precise monitoring of posttransplant hematopoietic chimerism is of utmost interest. Useful molecular methods for chimerism quantification after allogeneic transplantation, aimed at distinguishing precisely between donor's and recipient's cells, are PCR-based analyses of polymorphic DNA markers. Such analyses can be performed regardless of donor's and recipient's sex. Additionally, in patients after sex-mismatched allo-HSCT, fluorescent in situ hybridization (FISH) can be applied. Methods: We compared different techniques for analysis of posttransplant chimerism, namely FISH and PCR-based molecular methods with automated detection of fluorescent products in an ALFExpress DNA Sequencer (Pharmacia) or ABI 310 Genetic Analyzer (PE). We used Spearman correlation test. Results: We have found high correlation between results obtained from the PCR/ALF Express and PCR/ABI 310 Genetic Analyzer. Lower, but still positive correlations were found between results of FISH technique and results obtained using automated DNA sizing technology. Conclusions: All the methods applied enable a rapid and accurate detection of post-HSCT chimerism.
Poster presentation: NO-sensitive guanylyl cyclases (GC) are the principal receptors for nitric oxide (NO) and convert GTP into the second messenger cGMP. We showed that GC is prone to tyrosine phosphorylation in COS1 cells overexpressing the human holoenzyme. Similar results were obtained in PC12 cells and in rat aortic tissue slices. The major phosphorylation site was mapped to position 192 in the regulatory domain of the beta1 subunit. Tyrosine phosphorylation of GC was reduced in the presence of the inhibitors PP1 and PP2 indicating that Src-like kinases are critically involved in phosphorylation. Moreover, co-immunoprecipitation experiments revealed an interaction between Src and GC. To further analyse the relevance of this posttranslational modification we generated a phospho-specific antibody raised against pTyr192. This antibody clearly distinguishes between phosphorylated and non-phosphorylated GC and may be a powerful tool to analyse the subcellular localisation of the phosphorylated enzyme.
Poster presentation: NO-sensitive guanylyl cyclases (sGCs) are cytosolic receptors for nitric oxide (NO) catalyzing the conversion of GTP to cGMP. sGCs are obligate heterodimers composed of one alpha and beta subunit each. The allosteric mechanism of sGC activation via NO is well understood, however, our knowledge about alternative mechanisms such as protein-protein interactions regulating activity, availability, translocation and expression of sGC is rather limited. In a search by the yeast two-hybrid system using the catalytic domain of the alpha1 subunit as the bait, we have identified two structurally related proteins AGAP1 [1] and MRIP2 as novel sGC interacting proteins. MRIP2 is a multi-domain protein of 75 kDa comprising a single PH and ArfGAP domain each and two ankyrin repeats. Co-immunoprecipitation experiments using COS1 cells overexpressing both proteins demonstrated the interaction of MRIP2 with both subunits of the sGC alpha1beta1. Confocal microscopical analysis showed a prominent plasma membrane staining of MRIP2. This membrane association is mediated through an N-terminal myristoylation site and through binding of its PH domain to phospholipids such as phosphatidylinositol-3,5-bisphosphate (PI(3,5)P2). We hypothesize that MRIP2 may represent an acceptor protein for sGC that mediates recruitment of cytosolic sGC to the plasma membrane or other subcellular compartments.
Poster presentation NO-sensitive guanylyl cyclases (soluble guanylyl cyclase, sGC) are among the key regulators of intracellular cGMP concentration. The mechanisms underlying NO-mediated activation of sGC are quite well understood, however, little is known about the fine-tuning of sGC activity through alternative mechanisms such as protein phosphorylation. Several reports have demonstrated the reversible phosphorylation of sGC on serine/threonine residues, and it has been speculated, though not experimentally proven, that sGC might also be phosphorylated on tyrosine residues. Using broad-spectrum phosphatase inhibitors we were able to demonstrate tyrosine phosphorylation at Tyr192 of the beta 1 subunit of human sGC in COS1 cells. This residue forms part of a sequence segment (YEDL) representing a preferential binding site for SH2 domains of Src-like kinases. Pull-down assays and co-immunoprecipitation experiments showed that Src can indeed bind via its SH2 domain to pTyr192 of beta 1 indicating that tyrosine phosphorylation of sGC may be followed by recruitment of Src-like kinases to the phosphorylated beta 1 subunit. In support of this hypothesis, immunofluorescence studies showed a colocalization of overexpressed sGC and Src at the plasma membrane of COS1 and Hela cells. Together, our results point to an unexpected crosstalk between tyrosine kinase pathway(s) and the NO/cGMP signalling cascade which may result in translocation of the predominantly cytosolic sGC to the cytosolic face of the plasma membrane.
Severe acute respiratory syndrome (SARS) is a new infectious disease caused by a novel coronavirus that leads to deleterious pulmonary pathological features. Due to its high morbidity and mortality and widespread occurrence, SARS has evolved as an important respiratory disease which may be encountered everywhere in the world. The virus was identified as the causative agent of SARS due to the efforts of a WHO-led laboratory network. The potential mutability of the SARS-CoV genome may lead to new SARS outbreaks and several regions of the viral genomes open reading frames have been identified which may contribute to the severe virulence of the virus. With regard to the pathogenesis of SARS, several mechanisms involving both direct effects on target cells and indirect effects via the immune system may exist. Vaccination would offer the most attractive approach to prevent new epidemics of SARS, but the development of vaccines is difficult due to missing data on the role of immune system-virus interactions and the potential mutability of the virus. Even in a situation of no new infections, SARS remains a major health hazard, as new epidemics may arise. Therefore, further experimental and clinical research is required to control the disease.
Keywords: Severe Acute Respiratory Syndrome; SARS; coronavirus; molecular mechanisms; therapy; vaccination
Doğan, Selami ; Akbulut, Birkan ; Aybek, Tayfun ; Mierdl, Stefan ; Moritz, Anton ; Wimmer-Greinecker, Gerhard
Minimal invazif endoskopik giriflimlerin kalp cerrahisinde kullanımı ancak telemanipülatör sistemlerin kullanıma girmesiyle mümkün olmufltur. Bu çalıflmada total endoskopik revaskülarizasyon için kullanılan robotik destekli telemanipülasyon sistemleri gözden geçirilip, çalıflan ve duran kalpte uygulamalar derlenmifltir. Robotik cerrahi günümüzde halen geliflme aflamasındadır. Maliyetin yüksek oluflu ve sadece seçilmifl bir hasta grubunda uygulanabiliyor olması bu yeni tekni¤in en büyük sınırlayıcı faktörleridir. Ancak teknoloji üzerine çalıflmalar ve özellikle anastomoz tekniklerinin geliflmesiyle koroner revaskülarizasyon için bir alternatif olacaktır. Henüz istenen hedeflere ulaflılmamıflsa da gelecek umut vericidir.
Background: To investigate the occupational risk of tuberculosis (TB) infection in a low-incidence setting, data from a prospective study of patients with culture-confirmed TB conducted in Hamburg, Germany, from 1997 to 2002 were evaluated. Methods: M. tuberculosis isolates were genotyped by IS6110 RFLP analysis. Results of contact tracing and additional patient interviews were used for further epidemiological analyses. Results: Out of 848 cases included in the cluster analysis, 286 (33.7%) were classified into 76 clusters comprising 2 to 39 patients. In total, two patients in the non-cluster and eight patients in the cluster group were health-care workers. Logistic regression analysis confirmed work in the health-care sector as the strongest predictor for clustering (OR 17.9). However, only two of the eight transmission links among the eight clusters involving health-care workers had been detected previously. Overall, conventional contact tracing performed before genotyping had identified only 26 (25.2%) of the 103 contact persons with the disease among the clustered cases whose transmission links were epidemiologically verified. Conclusion: Recent transmission was found to be strongly associated with health-care work in a setting with low incidence of TB. Conventional contact tracing alone was shown to be insufficient to discover recent transmission chains. The data presented also indicate the need for establishing improved TB control strategies in health-care settings.
Minckwitz, Gunter von ; Harder, Sebastian ; Hövelmann, Sascha ; Jäger, Elke ; Batran, Salah-Eddin al- ; Loibl, Sibylle ; Atmaca, Akin ; Cimpoiasu, Christian ; Neumann, Antje ; Abera, Aklil ; Knuth, Alexander ; Kaufmann, Manfred ; Jäger, Dirk ; Maurer, Alexander B. ; Wels, Winfried
Introduction: ScFv(FRP5)-ETA is a recombinant antibody toxin with binding specificity for ErbB2 (HER2). It consists of an N-terminal single-chain antibody fragment (scFv), genetically linked to truncated Pseudomonas exotoxin A (ETA). Potent antitumoral activity of scFv(FRP5)-ETA against ErbB2-overexpressing tumor cells was previously demonstrated in vitro and in animal models. Here we report the first systemic application of scFv(FRP5)-ETA in human cancer patients.
Methods: We have performed a phase I dose-finding study, with the objective to assess the maximum tolerated dose and the dose-limiting toxicity of intravenously injected scFv(FRP5)-ETA. Eighteen patients suffering from ErbB2-expressing metastatic breast cancers, prostate cancers, head and neck cancer, non small cell lung cancer, or transitional cell carcinoma were treated. Dose levels of 2, 4, 10, 12.5, and 20 μg/kg scFv(FRP5)-ETA were administered as five daily infusions each for two consecutive weeks.
Results: No hematologic, renal, and/or cardiovascular toxicities were noted in any of the patients treated. However, transient elevation of liver enzymes was observed, and considered dose limiting, in one of six patients at the maximum tolerated dose of 12.5 μg/kg, and in two of three patients at 20 μg/kg. Fifteen minutes after injection, peak concentrations of more than 100 ng/ml scFv(FRP5)-ETA were obtained at a dose of 10 μg/kg, indicating that predicted therapeutic levels of the recombinant protein can be applied without inducing toxic side effects. Induction of antibodies against scFv(FRP5)-ETA was observed 8 days after initiation of therapy in 13 patients investigated, but only in five of these patients could neutralizing activity be detected. Two patients showed stable disease and in three patients clinical signs of activity in terms of signs and symptoms were observed (all treated at doses ≥ 10 μg/kg). Disease progression occurred in 11 of the patients.
Conclusion: Our results demonstrate that systemic therapy with scFv(FRP5)-ETA can be safely administered up to a maximum tolerated dose of 12.5 μg/kg in patients with ErbB2-expressing tumors, justifying further clinical development.
Schmidtko, Achim ; Tegeder, Irmgard ; Niederberger, Ellen ; Hofmann, Franz ; Ruth, Peter ; Geisslinger, Gerd
First paragraph (this article has no abstract) Persistent stimulation of nociceptors results in sensitization of nociceptive sensory neurons, which is associated with hyperalgesia and allodynia. The release of NO and subsequent synthesis of cGMP in the spinal cord are involved in this process. cGMP-dependent protein kinase I (PKG-I) has been suggested to act as a downstream target of cGMP, but its exact role in nociception hadn't been characterized yet. To further evaluate the NO/cGMP/PKG-I pathway in nociception we assessed the effects of PKG-I inhibiton and activaton in the rat formalin assay and analyzed the nociceptive behavior of PKG-I-/- mice. Open access article.
Vollmar, Horst Christian ; Waldmann, Uta-Maria ; Soennichsen, Andreas ; Schürer-Maly, Cornelia-Christine ; Gilbert, Katja ; Gensichen, Jochen
Kurzfassung Vortrag: Fächerübergreifende Lehre und verpflichtende Fortbildung (CME) für Ärzte erfordern innovative Lernmethoden. Eine Lösung wird teilweise in der Nutzung elektronischer Medien gesehen. Unklar ist jedoch, wie konkret eine Umsetzung in der Aus-, Fort- und Weiterbildung im Fach Allgemeinmedizin bisher erfolgte, welche Chancen und Möglichkeiten es gibt und wie die weitere Entwicklung aussehen kann. Um einen Überblick über die aktuellen E-Learning-Aktivitäten zu erhalten, formierte sich eine universitäts- und bundesländerübergreifende Initiative. In drei Phasen soll die Grundlage für die Entwicklung einer Strategie für die effektive Nutzung elektronischer Lehr- und Lernmedien für das Fach Allgemeinmedizin geschaffen werden: Phase 1 - Nationales Expertentreffen (Juli 2005 in Frankfurt): Diskussion von Erfahrungen, Problemen und Möglichkeiten des Einsatzes elektronischer Medien in der Allgemeinmedizin. Phase 2 - Gründung eines Netzwerks: Zusammenarbeit, Koordination und gegenseitige Unterstützung bei der Entwicklung von E-Learning-Modulen. Phase 3 - Evaluation von E-Learning in der Allgemeinmedizin: systematische qualitative und quantitative Untersuchungen. Im Rahmen eines Workshops sollen die Erfahrungen aus dem Expertentreffen mit Lehrenden und Fortbildenden aus anderen Fachbereichen diskutiert werden. Kooperationen über die Allgemeinmedizin hinaus können weitere Synergien schaffen. Der Workshop dient dem Austausch über Chancen und Limitationen entsprechender Angebote.
Neben dem alltagsbegrifflichen Verständnis der "Unternehmer", das deren Lagerbildung und Interessenhomogenität unterstellt, lassen sich bei genauerem Hinsehen immer schon interne Differenzierungen erkennen, erfassbar v.a. mit der Kategorie der "Kapitalfraktionen". Öffentliche politische Interventionen des "Unternehmerlagers" erscheinen wiederum auf den ersten Blick homogen. Rolf Schmucker zeigt empirisch, dass auch dieser Blick trügt.
Gensichen, Jochen ; Torge, Marion ; Peitz, Monika ; Wendt-Hermainski, Heike ; Beyer, Martin ; Rosemann, Thomas ; Krauth, Christian ; Raspe, Heiner ; Aldenhoff, Josef B. ; Gerlach, Ferdinand M.
Background: Depression is a disorder with high prevalence in primary health care and a significant burden of illness. The delivery of health care for depression, as well as other chronic illnesses, has been criticized for several reasons and new strategies to address the needs of these illnesses have been advocated. Case management is a patient-centered approach which has shown efficacy in the treatment of depression in highly organized Health Maintenance Organization (HMO) settings and which might also be effective in other, less structured settings. Methods/Design: PRoMPT (PRimary care Monitoring for depressive Patients Trial) is a cluster randomised controlled trial with General Practice (GP) as the unit of randomisation. The aim of the study is to evaluate a GP applied case-management for patients with major depressive disorder. 70 GPs were randomised either to intervention group or to control group with the control group delivering usual care. Each GP will include 10 patients suffering from major depressive disorder according to the DSM-IV criteria. The intervention group will receive treatment based on standardized guidelines and monthly telephone monitoring from a trained practice nurse. The nurse investigates the patient's status concerning the MDD criteria, his adherence to GPs prescriptions, possible side effects of medication, and treatment goal attainment. The control group receives usual care – including recommended guidelines. Main outcome measure is the cumulative score of the section depressive disorders (PHQ-9) from the German version of the Prime MD Patient Health Questionnaire (PHQ-D). Secondary outcome measures are the Beck-Depression-Inventory, self-reported adherence (adapted from Moriskey) and the SF-36. In addition, data are collected about patients' satisfaction (EUROPEP-tool), medication, health care utilization, comorbidity, suicide attempts and days out of work. The study comprises three assessment times: baseline (T0) , follow-up after 6 months (T1) and follow-up after 12 months (T2). Discussion: Depression is now recognized as a disorder with a high prevalence in primary care but with insufficient treatment response. Case management seems to be a promising intervention which has the potential to bridge the gap of the usually time-limited and fragmented provision of care. Case management has been proven to be effective in several studies but its application in the private general medical practice setting remains unclear.
Joos, Stefanie ; Rosemann, Thomas ; Heiderhoff, Marc ; Wensing, Michel ; Ludt, Sabine ; Gensichen, Jochen ; Kaufmann-Kolle, Petra ; Szecsenyi, Joachim
Background: Diabetes model projects in different regions of Germany including interventions such as quality circles, patient education and documentation of medical findings have shown improvements of HbA1c levels, blood pressure and occurrence of hypoglycaemia in before-after studies (without control group). In 2002 the German Ministry of Health defined legal regulations for the introduction of nationwide disease management programs (DMP) to improve the quality of care in chronically ill patients. In April 2003 the first DMP for patients with type 2 diabetes was accredited. The evaluation of the DMP is essential and has been made obligatory in Germany by the Fifth Book of Social Code. The aim of the study is to assess the effectiveness of DMP by example of type 2 diabetes in the primary care setting of two German federal states (Rheinland-Pfalz and Sachsen-Anhalt). Methods/Design: The study is three-armed: a prospective cluster-randomized comparison of two interventions (DMP 1 and DMP 2) against routine care without DMP as control group. In the DMP group 1 the patients are treated according to the current situation within the German-Diabetes-DMP. The DMP group 2 represents diabetic care within ideally implemented DMP providing additional interventions (e.g. quality circles, outreach visits). According to a sample size calculation a sample size of 200 GPs (each GP including 20 patients) will be required for the comparison of DMP 1 and DMP 2 considering possible drop-outs. For the comparison with routine care 4000 patients identified by diabetic tracer medication and age (> 50 years) will be analyzed. Discussion: This study will evaluate the effectiveness of the German Diabetes-DMP compared to a Diabetes-DMP providing additional interventions and routine care in the primary care setting of two different German federal states.
Kurzfassung eines Vortrags gehalten auf der GMA-Tagung vom 04.11.-06.11.2005 in Münster. Aufgrund wenig überzeugender Leistungen seiner Studierenden im vorklinischen Studienabschnitt hat der Fachbereich Medizin der Goethe-Universität zum Wintersemester 2001/02 eine völlig neue Studienordnung eingeführt, deren wesentliche Ziele das erfolgreiche Erlernen natur-, geistes- und sozialwissenschaftlicher Komponenten der normalen Biologie des Menschen mit optimaler Vorbereitung auf den klinischen Studienabschnitt binnen zweier Studienjahre und die Kompatibilität mit der ÄAppO und der ZÄAppO (jeweils alte und neue Version bzw. Vorentwurf) sind. Grundprinzipien des Curriculums sind: 1. Logisch aufeinander abgestimmte Themenblöcke, 2. Integration von makroskopischer und mikroskopischer Anatomie, 3. Interdisziplinärität der Themenblöcke, 4. Immanente Wiederholung von Lehrinhalten durch enge Verzahnung von Plenar- und Kleingruppenveranstaltungen, 5. Aufteilung der Studienanfänger in gemischte Gruppen nach den Kriterien Geschlecht, Herkunft und Studienfach, 6. Erlernen des Umgangs mit Patienten schon vom ersten Semester an durch "Anatomie am Lebenden"-Kurse und durch den Kurs Einführung in die klinische Medizin. Wichtigste Ergebnisse: Steigerung der Referenzgruppe beim Physikum von ca. 25% auf >60%, relativ guter Prüfungserfolg, hervorragende Sozialisation durch die frühe randomisierte Zuordnung in Kleingruppen und der sehr glatte Übergang in den klinischen Abschnitt des Medizin- bzw. Zahnmedizinstudiums.
Kongressbericht: Auf der Tagung der Deutschen Gesellschaft für Allgemeinmedizin und Familienmedizin e.V. (DEGAM) 2004 entstand die Idee, E-Learning-Aktivitäten in der Allgemeinmedizin sichtbar zu machen und zu bündeln. Ein Kongress sollte die allgemeinmedizinischen Vertreter aus Lehre und Forschung sowie Industrievertreter zusammenbringen, um das Spektrum der Möglichkeiten und laufende Projekte kennen zu lernen. Mit motivierten Referenten, über 60 aktiven Teilnehmern und einem positiven Feedback, kann der Kongress in Frankfurt am 8. und 9. Juli 2005 als erster dieser Art in Deutschland als erfolgreich bezeichnet werden.
The illusion of apparent motion can be induced when visual stimuli are successively presented at different locations. It has been shown in previous studies that motion-sensitive regions in extrastriate cortex are relevant for the processing of apparent motion, but it is unclear whether primary visual cortex (V1) is also involved in the representation of the illusory motion path. We investigated, in human subjects, apparent-motion-related activity in patches of V1 representing locations along the path of illusory stimulus motion using functional magnetic resonance imaging. Here we show that apparent motion caused a blood-oxygenation-level-dependent response along the V1 representations of the apparent-motion path, including regions that were not directly activated by the apparent-motion-inducing stimuli. This response was unaltered when participants had to perform an attention-demanding task that diverted their attention away from the stimulus. With a bistable motion quartet, we confirmed that the activity was related to the conscious perception of movement. Our data suggest that V1 is part of the network that represents the illusory path of apparent motion. The activation in V1 can be explained either by lateral interactions within V1 or by feedback mechanisms from higher visual areas, especially the motion-sensitive human MT/V5 complex.