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Cytoprotective mechanisms in cultured cardiomyocytes [chapter]

Hari S. Sharma, Joachim Stahl, Dirk Weisensee, Iris Löw-Friedrich
1996 Biochemical Mechanisms in Heart Function  
Tumor necrosis factor-c~ (TNF-a), a potent cytokine mainly secreted by macrophages exerts pleiotropic effects on different cell types. However, the intracellular mediators of its action are not yet well characterized. To get an insight into endogenous cytoprotective mechanisms, we developed an in vitro model based on cultured cardiomyocytes treated with TNF-c~ at which we examined gene expression of heat shock proteins (HSP-27, HSP-70 and ubiquitin). Cardiomyocytes were isolated from the hearts
more » ... of 18 day old fetal mice by enzymatic dissociation and grown in minimum essential medium containing 10% fetal calf serum. Spontaneously contractile cells were serum deprived for 24 h and treated with TNF-a (25 ng/ml) for 1, 2, 4, 6, 8, 12, and 24 h After each incubation, cells were processed to extract total proteins for Western and total RNA for Noi"thern blot analyses. TNF-ct induced arrhythmias and cessation of spontaneous contractions in a concentration and time dependent manner. Steady state (ubiquitin) or undetectable mRNA levels (HSP-27, HSP-70) were drastically induced (> 4 fold for all three genes vs untreated control cells) by TNF-c~, reaching maximal values between 6-8 h of stimulation. Thereafter, the expression of these stress genes declined but remained elevated as compared to control. By Western blot analysis, we found increased multiple bands of ubiquitin protein conjugates in TNF-c~ treated cells whereas no significant change in HSP-27 protein accumulation until 12 h was observed as compared to control. 24 h of TNF-ct incubation resulted in partial cellular necrosis. Our results indicate that TNF-c~ induces in cardiomyocytes transiently gene expression for cytoprotective molecules like HSP-27, HSP-70 and ubiquitin, suggesting these stress proteins to participate in subsequent defense mechanisms, for example in postischemic myocardial recovery. (Mol Cell Biochem 160/161:217-224, 1996)
doi:10.1007/978-1-4613-1279-6_28 fatcat:no2kqlj6inad3agkomh4toshte

Cytoprotective mechanisms in cultured cardiomyocytes

Hari S. Sharma, Joachim Stahl, Dirk Weisensee, Iris L�w-Friedrich
1996 Molecular and Cellular Biochemistry  
Tumor necrosis factor-c~ (TNF-a), a potent cytokine mainly secreted by macrophages exerts pleiotropic effects on different cell types. However, the intracellular mediators of its action are not yet well characterized. To get an insight into endogenous cytoprotective mechanisms, we developed an in vitro model based on cultured cardiomyocytes treated with TNF-c~ at which we examined gene expression of heat shock proteins (HSP-27, HSP-70 and ubiquitin). Cardiomyocytes were isolated from the hearts
more » ... of 18 day old fetal mice by enzymatic dissociation and grown in minimum essential medium containing 10% fetal calf serum. Spontaneously contractile cells were serum deprived for 24 h and treated with TNF-a (25 ng/ml) for 1, 2, 4, 6, 8, 12, and 24 h After each incubation, cells were processed to extract total proteins for Western and total RNA for Noi"thern blot analyses. TNF-ct induced arrhythmias and cessation of spontaneous contractions in a concentration and time dependent manner. Steady state (ubiquitin) or undetectable mRNA levels (HSP-27, HSP-70) were drastically induced (> 4 fold for all three genes vs untreated control cells) by TNF-c~, reaching maximal values between 6-8 h of stimulation. Thereafter, the expression of these stress genes declined but remained elevated as compared to control. By Western blot analysis, we found increased multiple bands of ubiquitin protein conjugates in TNF-c~ treated cells whereas no significant change in HSP-27 protein accumulation until 12 h was observed as compared to control. 24 h of TNF-ct incubation resulted in partial cellular necrosis. Our results indicate that TNF-c~ induces in cardiomyocytes transiently gene expression for cytoprotective molecules like HSP-27, HSP-70 and ubiquitin, suggesting these stress proteins to participate in subsequent defense mechanisms, for example in postischemic myocardial recovery. (Mol Cell Biochem 160/161:217-224, 1996)
doi:10.1007/bf00240052 pmid:8901476 fatcat:drz2dgvhefhi5jm23rueqwcfvi

Characterization of a gap-junctional intercellular communication (GJIC) assay using cigarette smoke

Ewald Roemer, Hans-Peter Lammerich, Lynda L. Conroy, Dirk Weisensee
2013 Toxicology Letters  
h i g h l i g h t s • Normal cells communicate by gap-junctional intercellular communication (GJIC). • Transformed cells show inhibition of GJIC. • We adapted and automated a GJIC assay. • Reproducibility and repeatability (for cigarette smoke) were 3.7% and 6.9%. • The assay could discriminate the promoting activity of different cigarette types. a b s t r a c t Inhibition of gap-junctional intercellular communication (GJIC) via exposure to various toxic substances has been implicated in tumor
more » ... romotion. In the present study, cigarette smoke total particulate matter (TPM), a known inhibitor of GJIC, were used to characterize a new GJIC screening assay in three independent experiments. The main features of this assay were automated fluorescence microscopy combined with non-invasive parachute technique. Rat liver epithelial cells (WB-F344) were stained with the fluorescent dye Calcein AM (acetoxymethyl) and exposed to TPM from the Kentucky Reference Cigarette 2R4F (a blend of Bright and Burley tobaccos) and from two single-tobacco cigarettes (Bright and Burley) for 3 h. Phorbol-12-myristate-13-acetate (TPA) was used as positive control and 0.5% dimethyl sulfoxide (DMSO) as solvent control. The transfer of dye to adjacent cells (percentage of stained cells) was used as a measure of cellular communication. A clear and reproducible dose-response of GJIC inhibition following TPM exposure was seen. Reproducibility and repeatability measurements for the 2R4F cigarette were 3.7% and 6.9%, respectively. The half-maximal effective concentration values were 0.34 ng/ml for TPA, 0.050 mg/ml for the 2R4F, 0.044 mg/ml for the Bright cigarette, and 0.060 mg/ml for the Burley cigarette. The assay was able to discriminate between the two single-tobacco cigarettes (P < 0.0001), and between the single-tobacco cigarettes and the 2R4F (P = 0.0008, 2R4F vs. Burley and P < 0.0001, 2R4F vs. Bright). Thus, this assay can be used to determine the activity of complex mixtures such as cigarette smoke with high throughput and high precision.
doi:10.1016/j.toxlet.2013.03.028 pmid:23558295 fatcat:q3o6kgx4uzcbdnsl4rm2yeoex4

Assessment of network perturbation amplitudes by applying high-throughput data to causal biological networks

Florian Martin, Ty M Thomson, Alain Sewer, David A Drubin, Carole Mathis, Dirk Weisensee, Dexter Pratt, Julia Hoeng, Manuel C Peitsch
2012 BMC Systems Biology  
High-throughput measurement technologies produce data sets that have the potential to elucidate the biological impact of disease, drug treatment, and environmental agents on humans. The scientific community faces an ongoing challenge in the analysis of these rich data sources to more accurately characterize biological processes that have been perturbed at the mechanistic level. Here, a new approach is built on previous methodologies in which high-throughput data was interpreted using prior
more » ... gical knowledge of cause and effect relationships. These relationships are structured into network models that describe specific biological processes, such as inflammatory signaling or cell cycle progression. This enables quantitative assessment of network perturbation in response to a given stimulus. Results: Four complementary methods were devised to quantify treatment-induced activity changes in processes described by network models. In addition, companion statistics were developed to qualify significance and specificity of the results. This approach is called Network Perturbation Amplitude (NPA) scoring because the amplitudes of treatment-induced perturbations are computed for biological network models. The NPA methods were tested on two transcriptomic data sets: normal human bronchial epithelial (NHBE) cells treated with the pro-inflammatory signaling mediator TNFα, and HCT116 colon cancer cells treated with the CDK cell cycle inhibitor R547. Each data set was scored against network models representing different aspects of inflammatory signaling and cell cycle progression, and these scores were compared with independent measures of pathway activity in NHBE cells to verify the approach. The NPA scoring method successfully quantified the amplitude of TNFα-induced perturbation for each network model when compared against NF-κB nuclear localization and cell number. In addition, the degree and specificity to which CDK-inhibition affected cell cycle and inflammatory signaling were meaningfully determined. Conclusions: The NPA scoring method leverages high-throughput measurements and a priori literature-derived knowledge in the form of network models to characterize the activity change for a broad collection of biological processes at high-resolution. Applications of this framework include comparative assessment of the biological impact caused by environmental factors, toxic substances, or drug treatments.
doi:10.1186/1752-0509-6-54 pmid:22651900 pmcid:PMC3433335 fatcat:ggi54obhdfd7lbc24tgawhlvi4

A Modular Cell-Type Focused Inflammatory Process Network Model for Non-Diseased Pulmonary Tissue

Jurjen W. Westra, Walter K. Schlage, Arnd Hengstermann, Stephan Gebel, Carole Mathis, Ty Thomson, Ben Wong, Vy Hoang, Emilija Veljkovic, Michael Peck, Rosemarie B. Lichtner, Dirk Weisensee (+4 others)
2013 Bioinformatics and Biology Insights  
Exposure to environmental stressors such as cigarette smoke (CS) elicits a variety of biological responses in humans, including the induction of inflammatory responses. These responses are especially pronounced in the lung, where pulmonary cells sit at the interface between the body's internal and external environments. We combined a literature survey with a computational analysis of multiple transcriptomic data sets to construct a computable causal network model (the Inflammatory Process
more » ... k (IPN)) of the main pulmonary inflammatory processes. The IPN model predicted decreased epithelial cell barrier defenses and increased mucus hypersecretion in human bronchial epithelial cells, and an attenuated pro-inflammatory (M1) profile in alveolar macrophages following exposure to CS, consistent with prior results. The IPN provides a comprehensive framework of experimentally supported pathways related to CS-induced pulmonary inflammation. The IPN is freely available to the scientific community as a resource with broad applicability to study the pathogenesis of pulmonary disease.
doi:10.4137/bbi.s11509 pmid:23843693 pmcid:PMC3700945 fatcat:gax26ypoc5bnvcf4yqkfjomhhq

Human bronchial epithelial cells exposed in vitro to cigarette smoke at the air-liquid interface resemble bronchial epithelium from human smokers

Carole Mathis, Carine Poussin, Dirk Weisensee, Stephan Gebel, Arnd Hengstermann, Alain Sewer, Vincenzo Belcastro, Yang Xiang, Sam Ansari, Sandra Wagner, Julia Hoeng, Manuel C. Peitsch
2013 American Journal of Physiology - Lung cellular and Molecular Physiology  
doi:10.1152/ajplung.00181.2012 pmid:23355383 pmcid:PMC3627940 fatcat:qofd34gd45eulbu3bbshoet5im

A Systems Biology Approach Reveals the Dose- and Time-Dependent Effect of Primary Human Airway Epithelium Tissue Culture after Exposure to Cigarette Smoke in Vitro

Carole Mathis, Stephan Gebel, Carine Poussin, Vincenzo Belcastro, Alain Sewer, Dirk Weisensee, Arnd Hengstermann, Sam Ansari, Sandra Wagner, Manuel C. Peitsch, Julia Hoeng
2015 Bioinformatics and Biology Insights  
To establish a relevant in vitro model for systems toxicology-based mechanistic assessment of environmental stressors such as cigarette smoke (CS), we exposed human organotypic bronchial epithelial tissue cultures at the air liquid interface (ALI) to various CS doses. Previously, we compared in vitro gene expression changes with published human airway epithelia in vivo data to assess their similarities. Here, we present a follow-up evaluation of these in vitro transcriptomics data, using
more » ... entary computational approaches and an integrated mRNA-microRNA (miRNA) analysis. The main cellular pathways perturbed by CS exposure were related to stress responses (oxidative stress and xenobiotic metabolism), inflammation (inhibition of nuclear factor-κB and the interferon gamma-dependent pathway), and proliferation/differentiation. Within post-exposure periods up to 48 hours, a transient kinetic response was observed at lower CS doses, whereas higher doses resulted in more sustained responses. In conclusion, this systems toxicology approach has potential for product testing according to "21st Century Toxicology".
doi:10.4137/bbi.s19908 pmid:25788831 pmcid:PMC4357630 fatcat:c2ucaxp6ujeatd7t4cwj2cft2y

DGfK-Präsident

Manfred Weisensee
2016 KN - Journal of Cartography and Geographic Information  
Manfred Weisensee, Hude Prasident der DGfK • Sektion Stuttgart Dieser Beitrag schliel3t an den Bericht von 2014 an, erschienen in den KN 2/2015, S. ]]6. Hohepunkt 2015 war der 63.  ...  Am gleichen Tag wird sion unter der Leitung von Dirk kehrsachsen seiner neuen Stadt die Sektion gemeinsam mit der Zellmer, die ein Arbeitstretfen in hat, weil er sich immer nur mit Society for News Design  ... 
doi:10.1007/bf03545298 fatcat:dc3adnmcafdjhgobwaci7iplwa

Neujahrsempfang der DGfK in Berlin

Mark Vetter
2015 KN - Journal of Cartography and Geographic Information  
Weisensee heißen die Besucher zum Neujahrsempfang willkommen. Foto: Mark Vetter der Universität Osnabrück gehalten.  ...  Horst Schöltier Vorstandsvorsitzender der Kartographie-Sti ftung Ravenstein Dirk Zellmer Vorsitzender der Jury • Neujahrsempf ang der DGf K in Berlin "Die Vereinten Nationen haben das internationale  ... 
doi:10.1007/bf03545510 fatcat:wny6dsgz6bfntfyv5sclq22xt4

Von der Kupferplatte zur interaktiven Karte

2015 KN - Journal of Cartography and Geographic Information  
Manfred Weisensee, Präsident der Jade Hochschule und Präsident der DGfK.  ...  Der Vorsitzende der Jury, Dirk ZelJmer, bei der Ravensteinpreis - vergabe Foto: Mark Vetter  ... 
doi:10.1007/bf03545492 fatcat:6xytxlqjlvbr3a755iec756sxe

DGfK-Sektion Mecklenburg-Vorpommern

Jörg Schaffer
2014 KN - Journal of Cartography and Geographic Information  
-Geograph Dirk Zellmer und Dipl.-Ing. Wolfgang Kolb als Vertreter der Jury. Dabei differenzierte die Jury Arbeiten von Auszubildenden und von Studieren- den.  ...  (DGfK) -Gesellschaft für Kartographie und Geomatik Manfred Weisensee, Hude Präsident der DGfK Besondere Wertschätzung brachte der Vortrag von Rechtsanwalt Claus Vester zum Ausdruck, der als Vertreter  ... 
doi:10.1007/bf03544107 fatcat:haa5uwgt75butopm3ak3xpu6n4

Sektion Halle-Leipzig

Thomas Chudy
2016 KN - Journal of Cartography and Geographic Information  
Manfred Weisensee, Hude Prasident der DGfK Bei dem Vortrag zu Uirmkarten galt es, seine person lichen Emotionen zu verorten, Foto: Th. Chudy tung.  ...  Mai Dirk Burghardt, TU Dresden, mit seinem groBe Beachtung findenden Vortrag liber die "emotional cartography -Erfassung, Modellierung und Visualisierung georeferenzierter Emotionen aus Photo-Metadaten  ... 
doi:10.1007/bf03545270 fatcat:rcvsdpw3fbasznr3plsyqnlvgi

Das Parlament im Globalisierungsprozess. Ein Desiderat in der Parlamentarismus- und Global Governance-Forschung

Marianne Beisheim, Achim Brunnengräber
2008 Zeitschrift für Internationale Beziehungen  
ZIB 1/2008 Desai, Meghnad 1998: Global Governance, in: Messner, Dirk (Hrsg.)  ...  In einem Gutachten für die Enquete-Kommission fordert Dirk ZIB 1/2008 Messner (2001) , dass Parlamente ihre Aufgabe als »Schnittstelleninstitution« des Mehrebenenregierens besser erkennen und umsetzen  ...  Generell fehlt bislang bei den Überlegungen zu Global Governance die »Perspektive der vergleichenden Regierungslehre fast vollständig« (Weisensee 2006: 674) .  ... 
doi:10.5771/0946-7165-2008-1-73 fatcat:liym7wuj5bhgji6ujwwmbtp7ly

New conceptual developments and technologies t XXV anniversary "Intergeo-2019" in Stutgart
Нові концептуальні розробки і технології на XXV ювілейному "INTERGEO-2019" в Штутгарті

I TREVOHO, Lviv Polytechnic National University, V ZADOROZHNYY, A KHOPTAR, Lviv Polytechnic National University, Lviv Polytechnic National University
2020 Modern achievements of geodesic science and industry  
Weisensee.  ...  Містосільський потікгеодані як основа для інноваційних програм мобільності, керівник Dirk Jacke (п'ять доповідей). 2.4.  ... 
doi:10.33841/1819-1339-1-39-06 fatcat:safxjkvebzcb3jh7p4meebgeyu

THE REMAKING OF GEOPOLITICAL SPACE AND INSTITUTIONAL TRANSFORMATIONS: THE CASE OF THE BALTIC REGION
Переформатирование геополитического пространства и институциональные трансформации (на примере Балтийского региона)

Pavel A. Barakhvostov
2021 Baltic Region  
Advocate Weisensee, on behalf of Haacken, asked the Court that "Mr.  ...  ," in Maurice Adams and Dirk Heirbaut (eds.), The Method and Culture of Comparative Law (Oxford: Hart Publishing, 2014) , 121-132. chapter 1 and suppress local customary laws.  ... 
doi:10.5922/2079-8555-2021-3-3 fatcat:w6g4ue34dvey3jlajz6xqqzadu
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