Fraedrich K.,Universitatsklinikum Hamburg Eppendorf |
Schrader J.,Universitatsklinikum Hamburg Eppendorf |
Ittrich H.,Universitatsklinikum Hamburg Eppendorf |
Keller G.,Universitatsklinikum Hamburg Eppendorf |
And 11 more authors.
Clinical Cancer Research | Year: 2012
Purpose: Aurora kinases play a crucial role in cell-cycle control. Uncontrolled expression of aurora kinases causes aneuploidy and tumor growth. As conservative treatment options for advanced gastroenteropancreatic neuroendocrine tumors (GEP-NET) are disappointing, aurora kinases may be an interesting target for novel therapeutic strategies. Experimental Design: Human GEP-NETs were tested for aurora kinase expression. The efficacy of the new aurora kinase inhibitor danusertib was evaluated in two human GEP-NET cell lines (BON1 and QGP) in vitro and in vivo. Results: The majority of ten insulinomas and all 33 nonfunctional pancreatic or midgut GEP-NETs expressed aurora A despite a mostly high degree of cell differentiation. Both human GEP-NET cell lines expressed aurora kinase A and B, and high Ser10 phosphorylation of histone H3 revealed increased aurora B activity. Remarkably, danusertib led to cell-cycle arrest and completely inhibited cell proliferation of the GEP-NET cells in vitro. Decreased phosphorylation of histone H3 indicated effective aurora B inhibition. In a subcutaneous murine xenograft model, danusertib significantly reduced tumor growth in vivo compared with controls or mice treated with streptozotocine/5-fluorouracil. As a consequence, decreased levels of tumor marker chromogranin A were found in mouse serum samples. In a newly developed orthotopic model for GEP-NET liver metastases by intrasplenic tumor cell transplantation, dynamic MRI proved significant growth inhibition of BON1- and QGP-derived liver metastases. Conclusions: These results show that danusertib may impose a new therapeutic strategy for aurora kinase expressing metastasized GEP-NETs. ©2012 AACR.
PubMed | Labor Lademannbogen, University College London, Albert Ludwigs University of Freiburg, University of Hamburg and University of Gottingen
Type: Journal Article | Journal: Gene | Year: 2016
Recently, gain-of-function (GOF) mutations in the gene encoding signal transducer and activator of transcription 1 (STAT1) have been associated with chronic mucocutaneous candidiasis (CMC). This case report describes a 10-year-old boy presenting with signs of common variable immunodeficiency (CVID), failure to thrive, impaired neurological development, and a history of recurrent mucocutaneous Candida infections. Sequencing of the STAT1 gene identified a heterozygous missense mutation in exon 7 encoding the STAT1 coiled-coil domain (c.514T>C, p.Phe172Leu). In addition to hypogammaglobulinemia with B-cell deficiency, and a low percentage of Th17 cells, immunological analysis of the patient revealed a marked depletion of forkhead-box P3(+)-expressing regulatory T cells (Tregs). In vitro stimulation of T cells from the patient with interferon- (IFN) and/or IFN resulted in a significantly increased expression of STAT1-regulated target genes such as MIG1, IRF1, MX1, MCP1/CCL2, IFI-56K, and CXCL10 as compared to IFN-treated cells from a healthy control, while no IFN/-mediated up-regulation of the FOXP3 gene was found. These data demonstrate that the STAT1 GOF mutation F172L, which results in impaired stability of the antiparallel STAT1 dimer conformation, is associated with inhibited Treg cell development and neurological symptoms.
Streichert T.,Universitatsklinikum Eppendorf Hamburg |
Otto B.,Universitatsklinikum Eppendorf Hamburg |
Schnabel C.,Universitatsklinikum Eppendorf Hamburg |
Nordholt G.,Universitatsklinikum Eppendorf Hamburg |
And 5 more authors.
Clinical Chemistry | Year: 2011
BACKGROUND: Pneumatic tube systems (PTSs) for the transport of blood samples are regaining popularity in medical centers after earlier reports that their use could introduce preanalytical distortions such as hemolysis and changes in blood gases. METHODS: We drew duplicate blood samples from 30 volunteers. One sample was hand transported, and the other sample was transported through a PTS together with a mini- data logger that provided continuous measurements of temperature, humidity, pressure, and acceleration. After transport the samples were analyzed at the same time. We looked for possible relationships of the transport method and the parameters measured by the data loggers with differences in hematological parameters, standard clinical chemistry analyses, blood coagulation, erythrocyte sedimentation rate, and blood gas analysis. RESULTS: There were no significant differences in temperature, humidity, and pressure between the methods of transport, but we observed significant differences in 3-axis accelerations. The combined effect of these forces could be described by the righttailed area under the vector sum acceleration distribution. Our data show that this area correlated with PTS speed and that PTS speed and the area under the curve exhibited a direct relation to the degree of hemolysis. CONCLUSIONS: Assessment of 3-axis acceleration by use of data loggers can be used to identify preanalytical deviations that result from the transportation of blood samples in PTSs. Our approach could be used for the evaluation and regular control of PTSs without the need for repeated blood drawing and laboratory analyses. © 2011 American Association for Clinical Chemistry.
Obermeier M.,Medizinisches Labor Dr Berg |
Pironti A.,Max Planck Institute for Informatics |
Berg T.,Medizinisches Labor Dr Berg |
Braun P.,PZB Aachen |
And 14 more authors.
Intervirology | Year: 2012
Background: Genotypic drug resistance testing provides essential information for guiding treatment in HIV-infected patients. It may either be used for identifying patients with transmitted drug resistance or to clarify reasons for treatment failure and to check for remaining treatment options. While different approaches for the interpretation of HIV sequence information are already available, no other available rules-based systems specifically have looked into the effects of combinations of drugs. HIV-GRADE (Genotypischer Resistenz Algorithmus Deutschland) was planned as a countrywide approach to establish standardized drug resistance interpretation in Germany and also to introduce rules for estimating the influence of mutations on drug combinations. The rules for HIV-GRADE are taken from the literature, clinical follow-up data and from a bioinformatics-driven interpretation system (geno2pheno [resistance]). HIV-GRADE presents the option of seeing the rules and results of other drug resistance algorithms for a given sequence simultaneously. Methods: The HIV-GRADE rules-based interpretation system was developed by the members of the HIV-GRADE registered society. For continuous updates, this expert committee meets twice a year to analyze data from various sources. Besides data from clinical studies and the centers involved, published correlations for mutations with drug resistance and genotype-phenotype correlation data information from the bioinformatic models of geno2pheno are used to generate the rules for the HIV-GRADE interpretation system. A freely available online tool was developed on the basis of the Stanford HIVdb rules interpretation tool using the algorithm specification interface. Clinical validation of the interpretation system was performed on the data of treatment episodes consisting of sequence information, antiretroviral treatment and viral load, before and 3 months after treatment change. Data were analyzed using multiple linear regression. Results: As the developed online tool allows easy comparison of different drug resistance interpretation systems, coefficients of determination (R 2) were compared for the freely available rules-based systems. HIV-GRADE (R 2 = 0.40), Stanford HIVdb (R 2 = 0.40), REGA algorithm (R 2 = 0.36) and ANRS (R 2 = 0.35) had a very similar performance using this multiple linear regression model. Conclusion: The performance of HIV-GRADE is comparable to alternative rules-based interpretation systems. While there is still room for improvement, HIV-GRADE has been made publicly available to allow access to our approach regarding the interpretation of resistance against single drugs and drug combinations. Copyright © 2012 S. Karger AG, Basel.
Herden U.,University of Hamburg |
Kromminga A.,University of Hamburg |
Hagel C.,Labor Lademannbogen |
Hartleb J.,Labor Lademannbogen |
And 3 more authors.
Therapeutic Drug Monitoring | Year: 2011
Pharmacodynamic drug monitoring might allow an improved use of immunosuppressive medication in transplant recipients. We assessed whether drug concentrations reflect the effect of cyclosporine (CsA) on expression of nuclear factor of activated T-cells-regulated cytokines. CsA drug concentrations and expression of interleukin-2, interferon-γ, and granulocyte-macrophage colony-stimulating factor in stimulated blood lymphocytes were determined predose (C0) and 2 hours after (C2) CsA intake in 20 de novo (less than 3 months) and 20 long-term (3 months to 10 years) liver transplant patients. The residual cytokine expression at C2 relative to C0 was calculated. Mean CsA C0 and C2 concentrations were 236 and 776 μg/L in de novo and 100 and 573 μg/L in long-term liver transplant patients, respectively. Two hours after CsA intake, the residual cytokine expression for all cytokines was comparable in both groups (de novo patients mean 16%; long-term patients mean 17%). CsA C2 concentrations showed a significant (P < 0.01) correlation with the residual cytokine expression of interleukin-2, interferon-γ, and granulocyte-macrophage colony-stimulating factor in both de novo and long-term patients, whereas CsA C0 concentrations did not. The data suggest that CsA C2 concentrations, but not C0 concentrations, reflect the effect of CsA on downregulation of cytokine expression in both de novo and long-term liver transplant patients. © 2011 by Lippincott Williams & Wilkins.
Ertl H.,Labor Lademannbogen |
Butte W.,Carl von Ossietzky University
Journal of Exposure Science and Environmental Epidemiology | Year: 2012
Semi-volatile chemicals like pesticides and polychlorinated biphenyls (PCB) tend to accumulate in house dust. This may result in residues of some parts per million (p.p.m.), closely associated with health impairments and diseases like cancer. To explain these associations, we must establish whether a relevant absorption from house dust into human organisms occurs, and most crucially the release of chemicals, that is, their bioaccessibility. Digestive as well as dermal bioaccessibilities were examined using in-vitro methods. On average, the digestive bioaccessibility was 40% for the pesticides and 60% for the PCB. The dermal penetration availability reached 60% for the pesticides and 70% for the PCB (percentages of the concentrations in the dust). Based on the bioaccessibility, an estimate of internal exposure was calculated and expressed as percentages of acceptable or tolerable daily intake (ADI/TDI) values. Exposure via the respiratory tract proved to be very low. Exposure via the digestive tract had maximum values of 4% for pesticides and 12% for PCB. Dermal exposure was much higher. Even for average concentrations in house dust (0.5 p.p.m.), children exposed to DDT and PCB showed up to 300% of the ADI/TDI values, and adults about 60%. With high concentrations of contaminants in house dust, the maximum doses absorbed through the skin reached 5000%. © 2012 Nature America, Inc. All rights reserved.
Arndt-Maric R.,Labor Lademannbogen
Clinical laboratory | Year: 2010
Dilated cardiomyopathy (DCM) is one of the leading causes of heart failure in the western world but there is still no specific and early diagnosis available. Besides a genetic predisposition and viral infections, autoimmune reactions play an important role in the pathogenesis of DCM. The beta1-adrenergic receptor (beta1-AR) has been described as the major target structure in autoimmune DCM. In this study a recombinant GST-beta1-AR fusion protein comprising the second extracellular loop was generated as a target for the analysis of autoantibodies in sera from 115 patients with different heart failure diseases (41 DCM, 30 non-ischemic secondary cardiomyopathy [NISCM], 44 coronary artery disease [CAD]). Sera were collected from a non-selected population of heart failure patients in consecutive order. Autoantibodies against the beta1-AR were detected in 37% of DCM, 30% of NISCM, and 36% of CAD patients but none of the controls were positive. Furthermore, our data show that cardiomyopathy patients with anti-beta1-AR antibodies are younger (54 years vs. 61 years [DCM], 53 years vs. 56 years [NISCM], 61 years vs. 61 years [CAD]. Regardless of diagnosis antibody-positive patients had lower EF levels (29% vs. 32%, p = 0.0001 [DCM]; 23% vs. 25%, p < 0.0001 [NISCM]; 23% vs. 25%, p < 0.0001 [CAD]) than the antibody-negative counterparts but, nevertheless, also lower NT-proBNP levels compared to antibody negative patients (567 pg/mL vs. 1296 pg/mL, p = 0.0005 [DCM]; 224 pg/mL vs. 1135 pg/mL, p = 0.0002 [NISCM]; 605 pg/mL vs. 940 pg/mL, p = 0.0005 [CAD]). We conclude that DCM patients should be further characterized and differentiated by the detection of autoantibodies against beta1-AR. Autoimmune DCM patients are younger compared with their non-autoimmune counterparts, possibly due to the autoimmune trigger of the disease or reflecting an early stage of the disease. Surprisingly, the autoimmune patients have worse clinical manifestations but show less excessive NT-proBNP levels. It is not clear yet, though, whether these autoantibodies have a direct impact on the NT-proBNP levels. Whether or not these data are a consequence of pathogenic antibodies has to be elucidated in further studies.
Jenderny J.,Labor Lademannbogen |
Schmidt W.,Labor Lademannbogen |
Bartsch O.,Johannes Gutenberg University Mainz
European Journal of Pediatrics | Year: 2010
Introduction: Low level of trisomy 13 mosaicism is a rare condition. In the present report, we describe a case of a 19-month-old boy with poor feeding, poor weight gain, mild dysmorphic features, mild muscular hypotonia, and speech delay. Discussion: Cytogenetic analysis on metaphases of lymphocytes revealed an 8% mosaic Robertsonian translocation trisomy 13 in the boy and a balanced Robertsonian translocation, 45,XX,der(13;14)(q10;q10), in his normal mother. Fluorescence in situ hybridization (FISH) on patient lymphocytes disclosed 4% of metaphases with a trisomy 13. The trisomy 13 mosaicism in metaphases could not be identified by interphase FISH. The percentage of three signals (4%) was within the standard deviation in diploid controls. Follow-up of the patient was performed at the age of 7 1/12 years, and in cells from buccal smear of the patient, trisomy 13 was detected in 11% of interphases analyzed that is a higher frequency. Uniparental disomy of chromosomes 13 and 14 were excluded in the boy, and therefore, his phenotypic abnormalities most likely were caused by the low level of trisomy 13 mosaicism. Conclusion: The detailed report of this patient described the infrequent occurrence of a low mosaic Robertsonian translocation trisomy 13. We suggest to study cases of low trisomy mosaicism preferentially using metaphase analyses rather than interphase FISH. Our case is helpful in further defining the phenotype of these patients. © 2009 Springer-Verlag.
Ingiliz P.,Medical Center for Infectious Diseases |
Krznaric I.,Medical Center for Infectious Diseases |
Stellbrink H..-J.,Study Center |
Knecht G.,Infectious Diseases Center |
And 9 more authors.
HIV Medicine | Year: 2014
Objectives: The incidence of sexually transmitted hepatitis C virus (HCV) reinfection is on the rise in HIV-infected men who have sex with men (MSM). Data on natural history of acute hepatitis C and possible factors associated with spontaneous clearance are limited. The aim of this study was to analyse the outcome of HCV reinfections in HIV-positive MSM. Methods: A retrospective analysis was carried out on patients with more than one sexually acquired HCV infection who were diagnosed at four major German HIV and hepatitis care centres. Reinfection was defined by genotype or phylogenetic clade switch, detectable HCV RNA after a sustained virological response (SVR) or after spontaneous clearance (SC). Results: In total, 48 HIV-positive MSM were identified with HCV reinfection, among them 11 with a third episode and one patient with four episodes. At the first episode, 43 and five patients had an SVR and SC, respectively. The second episode was accompanied by a genotype switch in 29 patients (60%). Whereas 30 and nine patients showed an SVR and SC, respectively, eight patients developed chronic hepatitis. Neither HCV genotype switch nor interleukin-28B genotype was associated with SC. However, SC rates at the second episode were higher for patients with SC at the first episode compared with patients without SC (60 vs. 14%, respectively; P=0.03). Two patients with SC at the first episode were reinfected with the same genotype. Conclusions: Multiple reinfections in HIV-infected MSM do occur, with or without genotype switch, and with prior SC of previous episodes. In this large case series, except for SC at the first episode, no factor was of value in clinical decision-making for early therapeutic intervention in acute HCV reinfection. Copyright © 2014 British HIV Association.
PubMed | Labor Lademannbogen
Type: Journal Article | Journal: Journal of exposure science & environmental epidemiology | Year: 2012
Semi-volatile chemicals like pesticides and polychlorinated biphenyls (PCB) tend to accumulate in house dust. This may result in residues of some parts per million (p.p.m.), closely associated with health impairments and diseases like cancer. To explain these associations, we must establish whether a relevant absorption from house dust into human organisms occurs, and most crucially the release of chemicals, that is, their bioaccessibility. Digestive as well as dermal bioaccessibilities were examined using in-vitro methods. On average, the digestive bioaccessibility was ~40% for the pesticides and ~60% for the PCB. The dermal penetration availability reached ~60% for the pesticides and ~70% for the PCB (percentages of the concentrations in the dust). Based on the bioaccessibility, an estimate of internal exposure was calculated and expressed as percentages of acceptable or tolerable daily intake (ADI/TDI) values. Exposure via the respiratory tract proved to be very low. Exposure via the digestive tract had maximum values of 4% for pesticides and 12% for PCB. Dermal exposure was much higher. Even for average concentrations in house dust (0.5p.p.m.), children exposed to DDT and PCB showed up to 300% of the ADI/TDI values, and adults about 60%. With high concentrations of contaminants in house dust, the maximum doses absorbed through the skin reached 5000%.