@article{AdamiDragstedEnigetal.1993,
  author    = {Adami, Hans-Olov and Dragsted, Lars and Enig, Bent and Hansen, Jens and Haraldsd{\´o}ttir, J{\´o}hanna and Hill, Michael J. and Holm, Lars Erik and Knudsen, Ib and Larsen, Jens-Jorgen and Lutz, Werner K. and Osler, Merete and Overvad, Kim and Sabroe, Svend and Sanner, Tore and Strube, Michael and Sorensen, Thorkild I. A. and Thorling, Eivind B.},
  title     = {Report from the working group on diet and cancer.},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-71601},
  year      = {1993},
  abstract  = {No abstract available.},
  subject      = {Krebs <Medizin>},
  language  = {en}
}
@article{LutzSchlatter1993,
  author    = {Lutz, Werner K. and Schlatter, Josef},
  title     = {The relative importance of mutagens and carcinogens in the diet.},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-86311},
  year      = {1993},
  abstract  = {Known mutagens and carcinogens in the dict were compiled and the risk of cancer was estimated on the basis of average exposure Ievels in Switzerland and carcinogenic potencies from rodent bioassays. The analysis showed that, except for a1cohol, the sum of all known dietary carcinogens could only explain a few percent of the cancer deaths attributed by epidemiologists to dietary factors. The discrepancy was explained by a "carcinogenicity" of excess macronutrients. This hypothesis was based on an evaluation of dietary restriction experiments in rats and mice, where a dramatic reducing effect on spontaneaus tumour formation was seen. From these experiments, a "carcinogenic potency" was deduced for food in excess (TD50 approximately 16 g/kg per day). Ovemutrition in Switzerland was converted into excess food intake and the cancer risk estimated on the basis ofthe TD50 value. The resulting risk of60,000 cases per one million lives wou1d aJlow to explain by overnutrition almost all "diet-related" cancer deaths in humans.},
  subject      = {Medizin},
  language  = {en}
}
@incollection{Lutz1991,
  author    = {Lutz, Werner K.},
  title     = {Dose-response relationships in chemical carcinogenesis: from DNA adducts to tumor incidence},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-71625},
  publisher      = {Universit{\"a}t W{\"u}rzburg},
  year      = {1991},
  abstract  = {Mechanistic possibilitles responsible for nonlinear shapes of the dose-response relationship in chemical carcinogenesis are discussed. (i) Induction and saturation of enzymatic activation and detoxification processes and of DNA repair affect the relationship between dose and steady-state DNA adduct Ievel; (ii) The fixation of DNA adducts in the form of mutations is accelerated by stimulation of the cell division, for Jnstance due to regenerative hyperplasia at cytotoxic dose Ievels; (iii) The rate of tumor formation results from a superposition of the rates of the individual steps. It can become exponential with dose if more than one step is accelerated by the DNA damage exerted by the genotoxic carcinogen. The strongly sigmoidal shapes often observed for dose-tumor incidence relationships in animal bioassays supports this analysis. A power of four for the dose in the su~linear part of the curve is the maximum observed (formaldehyde). In contrast to animal experiments, epidemiological data ln humans rarely show a slgnificant deviation from linearity. The discrepancy might be explained by the fact that a I arge nu mber of genes contribute to the overall sensitivity of an individual and to the respective heterogeneity within the human population. Mechanistic nonlinearities are flattened out in the presence of genetic and life-style factors which affect the sensitivity for the development of cancer. For a risk assessment, linear extrapolation from the high-dose lncidence to the spontaneaus rate can therefore be approprlate in a heterogeneous population even if the mechanism of action would result in a nonlinear shape of the dose-response curve in a homogeneaus population.},
  language  = {en}
}
@incollection{ShephardMeierLutz1991,
  author    = {Shephard, S. E. and Meier, I. and Lutz, Werner K.},
  title     = {Alkylating potency of nitrosated amino acids and peptides},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-86320},
  publisher      = {Universit{\"a}t W{\"u}rzburg},
  year      = {1991},
  abstract  = {Tbe alkylating potency of unstable N-nitrosamino acids and N-nitrosopeptides was investigated in vitro using 4-(para-nitrobenzyl)pyridine (NBP) as nucleophile. Of the amino acids, Met and those with an aromatic side chain were the most potent. The relative overall alkylating potency was 23:10:5:4:2:1: for Trp, Met, His, 1)rr, Phe and Gly, respectively. The homo-dipeptides were much more potent than the amino acids, with relative potencies of 400:110:100:8:3:1, for Trp-Trp, l)T-'I)T, Met-Met, Asp-Asp, Phe-Phe and Gly, respectively. In the one-phase reaction system (in which NBP is already present durlog the nitrosation reaction at acidic pH), all amino acids tested showed a second-order reaction for nitrite. In the two-phase system (in which NBP is added only after bringing the nitrosation reaction mixture to neutrality), all amino acids tested except one again showed a second-order reaction for nitrite (Phe, His, Asp and the dipeptide artiticial sweetener aspartame); only Met under these conditions bad a reaction order of one for nitrite. This could mean that nitrosation of the side chain of Metproduces a second N-nitroso product which is relatively stable in acid but reacts with NBP under neutral conditions. In the human stomach, this side-chain nitrosation might become more important than the reactions at the primary amino group, firstly because of the greater stability of the product(s) in acid and secondly because of the tirst-order reaction rate for nitrite. A decrease in nitrite concentration from the millimolar concentrations ofthe in-vitro assay to the micromolar concentrations in the stomach reduces the reaction rate by a factor of 1000 for the side-chain nitrosation, whereas a million-fold reduction will be observed for nitrosation of the amino group.},
  subject      = {Aminos{\"a}uren},
  language  = {en}
}
@phdthesis{Jarzina2022,
  author    = {Jarzina, Sebastian Oskar},
  title     = {Assessment of systemic toxicity in vitro using the Adverse Outcome Pathway (AOP) concept: nephrotoxicity due to receptor-mediated endocytosis and lysosomal overload and inhibition of mtDNA polymerase-ɣ as case studies},
  doi       = {10.25972/OPUS-26484},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-264842},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2022},
  abstract  = {The US National Research Council (NRC) report "Toxicity Testing in the 21st Century: A Vision and a strategy (Tox21)", published in 2007, calls for a complete paradigm shift in tox-icity testing. A central aspect of the proposed strategy includes the transition from apical end-points in in vivo studies to more mechanistically based in vitro tests. To support and facilitate the transition and paradigm shift in toxicity testing, the Adverse Outcome Pathway (AOP) concept is widely recognized as a pragmatic tool. As case studies, the AOP concept was ap-plied in this work to develop AOPs for proximal tubule injuries initiated by Receptor-mediated endocytosis and lysosomal overload and Inhibition of mtDNA polymerase-. These AOPs were used as a mechanistic basis for the development of in vitro assays for each key event (KE). To experimentally support the developed in vitro assays, proximal tubule cells from rat (NRK-52E) and human (RPTEC/TERT1) were treated with model compounds. To measure the dis-turbance of lysosomal function in the AOP - Receptor-mediated endocytosis and lysosomal overload, polymyxin antibiotics (polymyxin B, colistin, polymyxin B nonapeptide) were used as model compounds. Altered expression of lysosomal associated membrane protein 1/2 (LAMP-1/2) (KE1) and cathepsin D release from lysosomes (KE2) were determined by im-munofluorescence, while cytotoxicity (KE3) was measured using the CellTiter-Glo® cell via-bility assay. Importantly, significant differences in polymyxin uptake and susceptibility be-tween cell lines were observed, underlining the importance of in vitro biokinetics to determine an appropriate in vitro point of departure (PoD) for risk assessment. Compared to the in vivo situation, distinct expression of relevant transporters such as megalin and cubilin on mRNA and protein level in the used cell lines (RPTEC/TERT1 and NRK-52E) could not be con-firmed, making integration of quantitative in vitro to in vivo extrapolations (QIVIVE) neces-sary. Integration of QIVIVE by project partners of the University of Utrecht showed an im-provement in the modelled biokinetic data for polymyxin B. To assess the first key event, (KE1) Depletion of mitochondrial DNA, in the AOP - Inhibition of mtDNA polymerase-, a RT-qPCR method was used to determine the mtDNA copy number in cells treated with mod-el compounds (adefovir, cidofovir, tenofovir, adefovir dipivoxil, tenofovir disoproxil fumarate). Mitochondrial toxicity (KE2) was measured by project partners using the high-content imaging technique and MitoTracker® whereas cytotoxicity (KE3) was determined by CellTiter-Glo® cell viability assay. In contrast to the mechanistic hypothesis underlying the AOP - Inhibition of mtDNA polymerase-, treatment with model compounds for 24 h resulted in an increase rather than a decrease in mtDNA copy number (KE1). Only minor effects on mitochondrial toxicity (KE2) and cytotoxicity (KE3) were observed. Treatment of RPT-EC/TERT1 cells for 14 days showed only a slight decrease in mtDNA copy number after treatment with adefovir dipivoxil and tenofovir disoproxil fumarate, underscoring some of the limitations of short-term in vitro systems. To obtain a first estimation for risk assessment based on in vitro data, potential points of departure (PoD) for each KE were calculated from the obtained in vitro data. The most common PoDs were calculated such as the effect concentra-tion at which 10 \% or 20_\% effect was measured (EC10, EC20), the highest no observed effect concentration (NOEC), the lowest observed effect concentration (LOEC), the benchmark 10 \% (lower / upper) concentrations (BMC10, BMCL10, BMCU10) and a modelled non-toxic con-centration (NtC). These PoDs were then compared with serum and tissue concentrations de-termined from in vivo studies after treatment with therapeutic / supratherapeutic doses of the respective drugs in order to obtain a first estimate of risk based on in vitro data. In addition, AOPs were used to test whether the quantitative key event relationships between key events allow prediction of downstream effects and effects on the adverse outcome (AO) based on measurements of an early key event. Predictions of cytotoxicity from the mathematical rela-tionships showed good concordance with measured cytotoxicity after treatment with colistin and polymyxin b nonapeptide. The work also revealed uncertainties and limitations of the ap-plied strategy, which have a significant impact on the prediction and on a risk assessment based on in vitro results.},
  language  = {en}
}
@phdthesis{Kaestner2023,
  author    = {Kaestner, Alexandra Annika Nadine},
  title     = {Charakterisierung pharmakologischer Phosphoglykolatphosphatase-Inhibitoren},
  doi       = {10.25972/OPUS-27239},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-272394},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2023},
  abstract  = {In dieser Arbeit geht es um die Phosphoglykolatphosphatase (PGP), die als Phosphatase vom Haloazid Dehalogenase-Typ (HAD-Phosphatase) zu der ubiquit{\"a}r vorkommenden Superfamilie der HAD-Hydrolasen geh{\"o}rt. In der Literatur ist eine in vitro Phosphatase-Aktivit{\"a}t gegen{\"u}ber 2-Phospho-L-Laktat (2PL), 4-Phospho-D-Erythronat (4PE), Phosphoglykolat (PG) und Glycerol-3-Phosphat (G3P) beschrieben. 2PL und 4PE entstehen in Nebenreaktionen w{\"a}hrend der Glykolyse und hemmen bei Akkumulation die Glykolyse bzw. den Pentosephosphatweg. PG kann auch in einer Nebenreaktion w{\"a}hrend der Glykolyse oder im Rahmen der Reparatur von oxidativen DNA-Sch{\"a}den entstehen. G3P entsteht aus Dihydroxyacetonphosphat und bildet das Kohlenhydratger{\"u}st der Triacylglyceride (TAG). Zellul{\"a}re Studien konnten Hinweise auf die Regulierung des epidermalen wachstumsfaktor-(EGF-)induzierten Zytoskelettumbaus durch die PGP liefern und die Untersuchung von M{\"a}usen mit PGP-Inaktivierung zeigte einen Einfluss auf die Zellproliferation und embryonale Entwicklung. Die Regulation der PGP-Expression f{\"u}hrte zu Ver{\"a}nderungen im Kohlenhydrat- und Fettstoffwechsel. Die Untersuchung der PGP-Funktionen erfolgte bislang ausschließlich mit genetischen Ans{\"a}tzen. Aufgrund von m{\"o}glichen Kompensationsmechanismen und Off-Target-Effekten m{\"u}ssen genetische und pharmakologische Methoden als sich erg{\"a}nzende Ans{\"a}tze verstanden werden. Um die Funktionen der PGP besser zu verstehen, fokussiert sich die vorliegende Arbeit auf die gezielte pharmakologische PGP-Inhibition. In Vorarbeiten wurden 41.000 Molek{\"u}le gescreent und f{\"u}nf potentielle Inhibitoren identifiziert. Ziele dieser Arbeit waren zum einen die Implementierung der Inhibitor \# 1-Behandlung in der Zellkultur, zum anderen die Charakterisierung der PGP-Hemmung durch Inhibitor \# 48 und die Durchf{\"u}hrung erster Selektivit{\"a}tstestungen mit Inhibitor \# 48. Zusammenfassend kann festgehalten werden, dass Inhibitor \# 1 in der Lage ist, die endogene PGP in Zelllysaten der murinen spermatogonialen Zelllinie (GC1) zu hemmen. Unter bestimmten Bedingungen f{\"u}hrte die Inhibitor \# 1-Behandlung der GC1-Zellen zur Hemmung der PGP. Erste Analysen zellul{\"a}rer Inhibitoreffekte konnten eine Steigerung der TAG-Konzentration in behandelten GC1-Zellen nachweisen. Die PGP-Hemmung durch Inhibitor \# 48 wurde als unkompetitive Inhibition charakterisiert und es zeigten sich keine relevanten Inhibitoreffekte auf die HAD-Phosphatasen Magnesium-abh{\"a}ngige Phosphatase 1 (MDP1), Lysin-Histidin-Pyrophosphat-Phosphatase (LHPP) und Polynukleotidase 5'-Kinase/3'-Phosphatase (PnkP). Dagegen konnte eine Aktivit{\"a}tssteigerung von Phospho 2 beobachtet werden. Die vorliegende Arbeit liefert somit erste Erkenntnisse {\"u}ber die Anwendung des PGP-Inhibitors \# 1 in der Zellkultur und schafft die Grundlage f{\"u}r nachfolgende Untersuchungen mit Inhibitor \# 48. Weitere Experimente sind notwendig, die die Inhibitorbehandlung in der Zellkultur optimieren und die Selektivit{\"a}t weiter charakterisieren, um mithilfe der Inhibitoren neue Erkenntnisse {\"u}ber die physiologische und pathophysiologische Rolle der PGP gewinnen zu k{\"o}nnen.},
  subject      = {Phosphoglykolatphosphatase},
  language  = {de}
}
@phdthesis{Schuster2009,
  author    = {Schuster, Paul Xaver},
  title     = {Biotransformation of trans-1,1,1,3-tetrafluoropropene, 2,3,3,3-tetrafluoropropene and 1,2,3,3,3-pentafluoropropene},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-43716},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2009},
  abstract  = {trans-1,1,1,3-Tetrafluoropropene (HFO-1234ze) and 2,3,3,3-tetrafluoropropene (HFO-1234yf) are non-ozone-depleting fluorocarbon replacements with low global warming potentials and short atmospheric lifetimes. They are developed as foam blowing agent and refrigerant, respectively. Investigations on biotransformation in different test species and in vitro systems are required to assess possible health risks of human exposure and needed for commercial development. The biotransformation of HFO-1234ze and HFO-1234yf was therefore investigated after inhalation exposure. Male Sprague-Dawley rats were exposed to air containing 2 000; 10,000; or 50,000 ppm (n=5/concentration) HFO-1234ze or HFO-1234yf. Male B6C3F1 mice were only exposed to 50,000 ppm HFO-1234ze or HFO-1234yf. Due to lethality observed in a developmental study with rabbits after exposure to high concentrations of HFO-1234yf, the metabolic fate of the compound was tested by whole body inhalation exposure of female New Zealand White rabbits to air containing 2 000; 10,000; or 50,000 ppm (n=3/concentration) HFO-1234yf. All inhalation exposures were conducted for 6 h in a dynamic exposure chamber. After the end of the exposures, animals were individually housed in metabolic cages and urines were collected at 6 or 12 h intervals for 48 h (rats and mice) or 60 h (rabbits). For metabolite identification, urine samples were analyzed by 1H-coupled and 1H-decoupled 19F-NMR and by LC/MS-MS or GC/MS. Metabolites were identified by 19F-NMR chemical shifts, signal multiplicity, 1H-19F coupling constants and by comparison with synthetic reference compounds. Biotransformation of HFO-1234ze in rats exposed to 50,000 ppm yielded S-(3,3,3-trifluoro-trans-propenyl)mercaptolactic acid as the predominant metabolite which accounted for 66\% of all integrated 19F-NMR signals in urines. No 19F-NMR signals were found in spectra of rat urine samples collected after inhalation exposure to 2 000 or 10,000 ppm HFO-1234ze likely due to insufficient sensitivity. S-(3,3,3-Trifluoro-trans-propenyl)-L-cysteine, N-acetyl-S-(3,3,3-trifluoro-trans-propenyl)-L-cysteine, 3,3,3-trifluoropropionic acid and 3,3,3-trifluorolactic acid were also present as metabolites in urine samples of rats and mice at the 50,000 ppm level. A presumed amino acid conjugate of 3,3,3-trifluoropropionic acid was the major metabolite of HFO-1234ze in urine samples of mice exposed to 50,000 ppm and related to 18\% of total integrated 19F-NMR signals. Quantitation of three metabolites in urines of rats and mice was performed, using LC/MS-MS or GC/MS. The quantified amounts of the metabolites excreted with urine in both mice and rats, suggest only a low extent (<<1\% of dose received) of biotransformation of HFO-1234ze and 95\% of all metabolites were excreted within 18 h after the end of the exposures (t1/2 approx. 6 h). Due to its low boiling point of \&\#8722;22 °C, most of the inhaled HFO-1234ze is expected to be readily exhaled. Moreover, steric and electronic factors may decrease the reactivity of the parent compound with soft nucleophiles such as glutathione. The obtained results suggest that HFO-1234ze is subjected to an addition-elimination reaction with glutathione and to a cytochrome P450-mediated epoxidation at low rates. The extent of a direct addition reaction of HFO-1234ze with glutathione is negligible, compared to that of the observed addition-elimination reaction. The results of in vivo testing of HFO-1234ze could not be supported by in vitro investigations, since HFO-1234ze was not metabolized in incubations with either liver microsomes or subcellular fractions from rat and human. Regarding the structures delineated in the biotransformation scheme of HFO-1234ze, 1,1,1,3-tetrafluoroepoxypropane and 3,3,3-trifluoropropionic acid are toxic intermediates which, however, are not supposed to display toxicity in the species after exposure to HFO-1234ze, due to the low extent of formation and an efficient detoxification of the epoxide by hydrolysis and glutathione conjugation. The findings of biotransformation of HFO-1234ze in rats and mice correlate with the absence of adverse effects in the toxicity testings and indicate their innocuousness to a human exposure. Biotransformation of HFO-1234yf yielded N-acetyl-S-(3,3,3-trifluoro-2-hydroxypropanyl)-L-cysteine as predominat metabolite which accounted for approx. 44, 90 and 32\% (50,000 ppm) of total 19F-NMR signal intensities in urine samples from rabbits, rats and mice, respectively. S-(3,3,3-Trifluoro-2-hydroxypropanyl)mercaptolactic acid and the sulfoxides of mercapturic acid and mercaptolactic acid S-conjugate were identified as minor metabolites of HFO-1234yf in urine samples from rabbits, rats and mice, whereas trifluoroacetic acid, 3,3,3-trifluorolactic acid and 3,3,3-trifluoro-1-hydroxyacetone were present as minor metabolites only in urine samples from rats and mice. The absence of these metabolites in rabbit urine samples...},
  subject      = {Biotransformation},
  language  = {en}
}
@article{JeanclosSchloetzerHadameketal.2022,
  author    = {Jeanclos, Elisabeth and Schl{\"o}tzer, Jan and Hadamek, Kerstin and Yuan-Chen, Natalia and Alwahsh, Mohammad and Hollmann, Robert and Fratz, Stefanie and Yesilyurt-Gerhards, Dilan and Frankenbach, Tina and Engelmann, Daria and Keller, Angelika and Kaestner, Alexandra and Schmitz, Werner and Neuenschwander, Martin and Hergenr{\"o}der, Roland and Sotriffer, Christoph and von Kries, Jens Peter and Schindelin, Hermann and Gohla, Antje},
  title     = {Glycolytic flux control by drugging phosphoglycolate phosphatase},
  series = {Nature Communications},
  volume    = {13},
  journal   = {Nature Communications},
  number    = {1},
  doi       = {10.1038/s41467-022-34228-2},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-300928},
  year      = {2022},
  abstract  = {Targeting the intrinsic metabolism of immune or tumor cells is a therapeutic strategy in autoimmunity, chronic inflammation or cancer. Metabolite repair enzymes may represent an alternative target class for selective metabolic inhibition, but pharmacological tools to test this concept are needed. Here, we demonstrate that phosphoglycolate phosphatase (PGP), a prototypical metabolite repair enzyme in glycolysis, is a pharmacologically actionable target. Using a combination of small molecule screening, protein crystallography, molecular dynamics simulations and NMR metabolomics, we discover and analyze a compound (CP1) that inhibits PGP with high selectivity and submicromolar potency. CP1 locks the phosphatase in a catalytically inactive conformation, dampens glycolytic flux, and phenocopies effects of cellular PGP-deficiency. This study provides key insights into effective and precise PGP targeting, at the same time validating an allosteric approach to control glycolysis that could advance discoveries of innovative therapeutic candidates.},
  language  = {en}
}
@phdthesis{Lotz2023,
  author    = {Lotz, Arietta Lucia},
  title     = {Eine in-vitro-Untersuchung des Einflusses von Angiotensin II und Sulforaphan auf die Modulation des oxidativen Stresses anhand der NFκB- und Nrf 2-Aktivit{\"a}t in LLC-PK1 Zellen},
  doi       = {10.25972/OPUS-31057},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-310573},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2023},
  abstract  = {Ausgangspunkt der Arbeit ist die klinische Beobachtung, dass Patienten mit arteriellem Hypertonus vermehrt Nierenerkrankungen entwickeln. Dabei zeigten sich in der Subgruppenanalyse vor allem erh{\"o}hte Inzidenzen der Niereninsuffizienz und der Nierenzellkarzinome. Als m{\"o}glicher Pathomechanismus steht das Renin-Angiotensin-Aldosteron-System (RAAS-System) im Vordergrund. Dabei wird postuliert, dass erh{\"o}hte Angiotensin II-Spiegel zu einem Missverh{\"a}ltnis zwischen den Oxidations- und Reduktionspartnern in der Zelle f{\"u}hren, wodurch sich das oxidative Potential der Zelle {\"a}ndert, und es vermehrt zur Bildung von Radikalen (ROS) kommt, die meist ungepaarte Elektronen in der Valenzschale oder instabile Verbindungen enthalten, wodurch sie besonders reaktionsfreudig mit Proteinen, Lipiden, Kohlenhydraten und auch der DNA interagieren. In der Folge kommt es zu DNA-Ver{\"a}nderungen in Form von Doppel- oder Einzelstrangbr{\"u}chen, DNA-Protein-Crosslinks, Basenmodifikationen und Basenverlusten, wodurch sich ein hohes mutagenes Potential ergibt. Dieser Ansatz zur Pathophysiologie best{\"a}tigte sich auch an den hier verwendeten porkinen Nierenzellmodell. Dabei zeigte sich nicht nur eine Ver{\"a}nderung der genomischen Stabilit{\"a}t nach Exposition gegen{\"u}ber erh{\"o}hten Angiotensin II-Spiegeln, sondern auch eine Ver{\"a}nderung der DNA in Abh{\"a}ngigkeit von der Expositionsdauer der Zellen. Als n{\"a}chster Schritt konnte die Modulation der Transkriptionsfaktoren Nrf 2 und NF-κB durch die Behandlung mit Angiotensin II und Sulforaphan nachgewiesen werden. Bei der Behandlung mit Sulforaphan ließ sich eine Nrf 2-Induktion nachweisen mit vermehrter Expression von antioxidativen und detoxifizierender Enzyme. Weiterhin zeigte sich im Rahmen der Behandlung erniedrigte NF-κB-Level. Bei der Modulation durch Angiotensin II stellte sich zun{\"a}chst ein signifikant erniedrigtes Level an Nrf 2 in den Zellen dar, das im Verlauf von 24 Stunden anstieg und konsekutiv ließ sich eine maximale Proteinexpression zwischen 24 und 48 Stunden messen. Weiterhin wiesen die Zellen, die mit Angiotensin II behandelt wurden, erh{\"o}hte NF-κB Mengen/Zelle auf. Zudem zeigte sich der Einfluss erh{\"o}hter Glucosekonzentrationen auf eine progrediente genomischen Instabilit{\"a}t, die Ver{\"a}nderung der Transkriptionsfaktoren mit erh{\"o}hter Nrf 2-Induktion und mit Deregulation des Transkriptionsfaktors NF-κB wurde durch die Behandlung mit Sulforaphan nachgewiesen. Aufgrund dieser Rolle in der Tumorgenese sind mittlerweile einige Bestandteile des NF-κB- und des Nrf 2-Signalweges und auch Nrf 2-Aktivatoren wie Sulforaphan wichtige Zielstrukturen f{\"u}r die Entwicklung neuer Medikamente und Therapieoptionen. Besonders zeigt sich hierbei die Wichtigkeit bei Diabetes induzierten kardiovaskul{\"a}ren Folgesch{\"a}den mit fr{\"u}hzeitiger medikament{\"o}ser Behandlung.},
  subject      = {Oxidativer Stress},
  language  = {de}
}
@phdthesis{Schott2021,
  author    = {Schott, Lea Marie},
  title     = {In vitro Untersuchung zur Genotoxizit{\"a}t ausgew{\"a}hlter Pyrrolizidinalkaloide},
  doi       = {10.25972/OPUS-24171},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-241716},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2021},
  abstract  = {Pyrrolizidinalkaloide (PA) sind sekund{\"a}re Pflanzenstoffe, welche {\"u}ber Nahrungsmittel in den menschlichen Organismus gelangen k{\"o}nnen. Zahlreiche Studien belegen, dass PA in der Leber verstoffwechselt und dabei in aktive genotoxische Metabolite umgewandelt werden. Diese verursachen vor allem in der Leber zellul{\"a}re Sch{\"a}den, was sich klinisch in Form einer hepatischen ven{\"o}sen okklusiven Leberkrankheit, aber auch in der Entstehung von Tumoren zeigt. Die vorliegende Arbeit testet das genotoxische Potential der drei PA Lasiocarpin, Senecionin und Seneciphyllin anhand der Leberzelllinie Huh6 mit Hilfe des Mikrokerntests. Dar{\"u}ber hinaus wird die Wirkung von Lasiocarpin auf den intrazellul{\"a}ren Glutathion-Gehalt, die Superoxidproduktion und das mitochondriale Membranpotential analysiert. Zudem werden sowohl der eventuell negative Einfluss einer Glutathion Depletion, als auch die m{\"o}glicherweise sch{\"u}tzenden Effekte des pflanzlichen Antioxidans Delphinidin in Bezug auf die Genotoxizit{\"a}t von Lasiocarpin untersucht. Es konnte gezeigt werden, dass alle drei ausgew{\"a}hlten PA einen signifikanten Anstieg der Mikrokernfrequenz bewirken.Unsere Messungen zeigten f{\"u}r Lasiocarpin eine dezente Reduktion des Glutathion Gehalts. Dagegen f{\"u}hrte eine Glutathion-Depletion in den Huh6 Zellen zu keiner Steigerung der Genotoxizit{\"a}t von Lasiocarpin. In Kombination mit dem Antioxidans Delphinidin zeigte sich f{\"u}r Lasiocarpin eine signifikante Reduktion der Mikrokernfrequenz. Abschließend ist anzumerken, dass in Zukunft vor allem die Wechselwirkung der PA untereinander und mit anderen (Pflanzen-)bestandteilen f{\"u}r eine verbesserte Risikoabsch{\"a}tzung der PA-Exposition untersucht werden sollte.},
  subject      = {Pyrrolizidinalkaloide},
  language  = {de}
}
@article{SchoeneggeGallionPicardetal.2017,
  author    = {Sch{\"o}negge, Anne-Marie and Gallion, Jonathan and Picard, Louis-Philippe and Wilkins, Angela D. and Le Gouill, Christian and Audet, Martin and Stallaert, Wayne and Lohse, Martin J. and Kimmel, Marek and Lichtarge, Olivier and Bouvier, Michel},
  title     = {Evolutionary action and structural basis of the allosteric switch controlling β\(_2\)AR functional selectivity},
  series = {Nature Communications},
  volume    = {8},
  journal   = {Nature Communications},
  doi       = {10.1038/s41467-017-02257-x},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-172268},
  year      = {2017},
  abstract  = {Functional selectivity of G-protein-coupled receptors is believed to originate from ligand-specific conformations that activate only subsets of signaling effectors. In this study, to identify molecular motifs playing important roles in transducing ligand binding into distinct signaling responses, we combined in silico evolutionary lineage analysis and structure-guided site-directed mutagenesis with large-scale functional signaling characterization and non-negative matrix factorization clustering of signaling profiles. Clustering based on the signaling profiles of 28 variants of the β\(_2\)-adrenergic receptor reveals three clearly distinct phenotypical clusters, showing selective impairments of either the Gi or βarrestin/endocytosis pathways with no effect on Gs activation. Robustness of the results is confirmed using simulation-based error propagation. The structural changes resulting from functionally biasing mutations centered around the DRY, NPxxY, and PIF motifs, selectively linking these micro-switches to unique signaling profiles. Our data identify different receptor regions that are important for the stabilization of distinct conformations underlying functional selectivity.},
  language  = {en}
}
@article{WernerWakabayashiBaueretal.2018,
  author    = {Werner, Rudolf and Wakabayashi, Hiroshi and Bauer, Jochen and Sch{\"u}tz, Claudia and Zechmeister, Christina and Hayakawa, Nobuyuki and Javadi, Mehrbod S. and Lapa, Constantin and Jahns, Roland and Erg{\"u}n, S{\"u}leyman and Jahns, Valerie and Higuchi, Takahiro},
  title     = {Longitudinal \(^{18}\)F-FDG PET imaging in a Rat Model of Autoimmune Myocarditis},
  series = {European Heart Journal Cardiovascular Imaging},
  journal   = {European Heart Journal Cardiovascular Imaging},
  issn      = {2047-2404},
  doi       = {10.1093/ehjci/jey119},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-165601},
  pages     = {1-8},
  year      = {2018},
  abstract  = {Aims: Although mortality rate is very high, diagnosis of acute myocarditis remains challenging with conventional tests. We aimed to elucidate the potential role of longitudinal 2-Deoxy-2-\(^{18}\)F-fluoro-D-glucose (\(^{18}\)F-FDG) positron emission tomography (PET) inflammation monitoring in a rat model of experimental autoimmune myocarditis. Methods and results: Autoimmune myocarditis was induced in Lewis rats by immunizing with porcine cardiac myosin emulsified in complete Freund's adjuvant. Time course of disease was assessed by longitudinal \(^{18}\)F-FDG PET imaging. A correlative analysis between in- and ex vivo \(^{18}\)F-FDG signalling and macrophage infiltration using CD68 staining was conducted. Finally, immunohistochemistry analysis of the cell-adhesion markers CD34 and CD44 was performed at different disease stages determined by longitudinal \(^{18}\)F-FDG PET imaging. After immunization, myocarditis rats revealed a temporal increase in 18F-FDG uptake (peaked at week 3), which was followed by a rapid decline thereafter. Localization of CD68 positive cells was well correlated with in vivo \(^{18}\)F-FDG PET signalling (R\(^2\) = 0.92) as well as with ex vivo 18F-FDG autoradiography (R\(^2\) = 0.9, P < 0.001, respectively). CD44 positivity was primarily observed at tissue samples obtained at acute phase (i.e. at peak 18F-FDG uptake), while CD34-positive staining areas were predominantly identified in samples harvested at both sub-acute and chronic phases (i.e. at \(^{18}\)F-FDG decrease). Conclusion: \(^{18}\)F-FDG PET imaging can provide non-invasive serial monitoring of cardiac inflammation in a rat model of acute myocarditis.},
  subject      = {Myokarditis},
  language  = {en}
}
@article{BreitenbachLorenzDandekar2019,
  author    = {Breitenbach, Tim and Lorenz, Kristina and Dandekar, Thomas},
  title     = {How to steer and control ERK and the ERK signaling cascade exemplified by looking at cardiac insufficiency},
  series = {International Journal of Molecular Sciences},
  volume    = {20},
  journal   = {International Journal of Molecular Sciences},
  number    = {9},
  issn      = {1422-0067},
  doi       = {10.3390/ijms20092179},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-285164},
  year      = {2019},
  abstract  = {Mathematical optimization framework allows the identification of certain nodes within a signaling network. In this work, we analyzed the complex extracellular-signal-regulated kinase 1 and 2 (ERK1/2) cascade in cardiomyocytes using the framework to find efficient adjustment screws for this cascade that is important for cardiomyocyte survival and maladaptive heart muscle growth. We modeled optimal pharmacological intervention points that are beneficial for the heart, but avoid the occurrence of a maladaptive ERK1/2 modification, the autophosphorylation of ERK at threonine 188 (ERK\(^{Thr188}\) phosphorylation), which causes cardiac hypertrophy. For this purpose, a network of a cardiomyocyte that was fitted to experimental data was equipped with external stimuli that model the pharmacological intervention points. Specifically, two situations were considered. In the first one, the cardiomyocyte was driven to a desired expression level with different treatment strategies. These strategies were quantified with respect to beneficial effects and maleficent side effects and then which one is the best treatment strategy was evaluated. In the second situation, it was shown how to model constitutively activated pathways and how to identify drug targets to obtain a desired activity level that is associated with a healthy state and in contrast to the maleficent expression pattern caused by the constitutively activated pathway. An implementation of the algorithms used for the calculations is also presented in this paper, which simplifies the application of the presented framework for drug targeting, optimal drug combinations and the systematic and automatic search for pharmacological intervention points. The codes were designed such that they can be combined with any mathematical model given by ordinary differential equations.},
  language  = {en}
}
@article{SchihadaVandenabeeleZabeletal.2018,
  author    = {Schihada, Hannes and Vandenabeele, Sylvie and Zabel, Ulrike and Frank, Monika and Lohse, Martin J. and Maiellaro, Isabella},
  title     = {A universal bioluminescence resonance energy transfer sensor design enables high-sensitivity screening of GPCR activation dynamics},
  series = {Communications Biology},
  volume    = {1},
  journal   = {Communications Biology},
  number    = {105},
  doi       = {10.1038/s42003-018-0072-0},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-228592},
  pages     = {1-8},
  year      = {2018},
  abstract  = {G-protein-coupled receptors (GPCRs) represent one of the most important classes of drug targets. The discovery of new GCPR therapeutics would greatly benefit from the development of a generalizable high-throughput assay to directly monitor their activation or de-activation. Here we screened a variety of labels inserted into the third intracellular loop and the C-terminus of the alpha(2 Lambda)-adrenergic receptor and used fluorescence (FRET) and bioluminescence resonance energy transfer (BRET) to monitor ligand-binding and activation dynamics. We then developed a universal intramolecular BRET receptor sensor design to quantify efficacy and potency of GPCR ligands in intact cells and real time. We demonstrate the transferability of the sensor design by cloning beta(2)-adrenergic and PTH1-receptor BRET sensors and monitored their efficacy and potency. For all biosensors, the Z factors were well above 0.5 showing the suitability of such design for microtiter plate assays. This technology will aid the identification of novel types of GPCR ligands.},
  language  = {en}
}
@article{IlinKulmanovNersesyanetal.2015,
  author    = {Ilin, Alexander and Kulmanov, Murat and Nersesyan, Armen and Stopper, Helga},
  title     = {Genotoxic activity of the new pharmaceutical FS-1 in Salmonella/microsome test and mouse lymphoma L5178Y cells},
  series = {Journal of BUON},
  volume    = {20},
  journal   = {Journal of BUON},
  number    = {2},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-143769},
  pages     = {595-601},
  year      = {2015},
  abstract  = {Purpose: The purpose of this study was to determine possible genotoxic effects of a new very promising antibacterial/ antiviral drug FS-1. Methods: The drug was tested in TA98, TA100, TA102, TA 1535 and TA1537 strains of Salmonella (Ames test) with and without metabolic activation, and also in mouse lymphoma L5178Y cells by means of micronucleus and comet assays. In microbes the drug was tested at concentrations up to 500 \(\mu\)g/plate and in mouse lymphoma cells up to 2,000 \(\mu\)g/ml. Results: In both test-systems in all experiments completely negative results were obtained although FS-1 was tested at maximum tolerated doses. Conclusions: The drug is not genotoxic. This is advantageous because many antibacterial/antiviral drugs possess such activity.},
  language  = {en}
}
@article{MallyJarzina2022,
  author    = {Mally, Angela and Jarzina, Sebastian},
  title     = {Mapping adverse outcome pathways for kidney injury as a basis for the development of mechanism-based animal-sparing approaches to assessment of nephrotoxicity},
  series = {Frontiers in Toxicology},
  volume    = {4},
  journal   = {Frontiers in Toxicology},
  issn      = {2673-3080},
  doi       = {10.3389/ftox.2022.863643},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-284405},
  year      = {2022},
  abstract  = {In line with recent OECD activities on the use of AOPs in developing Integrated Approaches to Testing and Assessment (IATAs), it is expected that systematic mapping of AOPs leading to systemic toxicity may provide a mechanistic framework for the development and implementation of mechanism-based in vitro endpoints. These may form part of an integrated testing strategy to reduce the need for repeated dose toxicity studies. Focusing on kidney and in particular the proximal tubule epithelium as a key target site of chemical-induced injury, the overall aim of this work is to contribute to building a network of AOPs leading to nephrotoxicity. Current mechanistic understanding of kidney injury initiated by 1) inhibition of mitochondrial DNA polymerase γ (mtDNA Polγ), 2) receptor mediated endocytosis and lysosomal overload, and 3) covalent protein binding, which all present fairly well established, common mechanisms by which certain chemicals or drugs may cause nephrotoxicity, is presented and systematically captured in a formal description of AOPs in line with the OECD AOP development programme and in accordance with the harmonized terminology provided by the Collaborative Adverse Outcome Pathway Wiki. The relative level of confidence in the established AOPs is assessed based on evolved Bradford-Hill weight of evidence considerations of biological plausibility, essentiality and empirical support (temporal and dose-response concordance).},
  language  = {en}
}
@phdthesis{Reiser2023,
  author    = {Reiser, Pia},
  title     = {Das Adverse Outcome Pathway (AOP) - Konzept als Grundlage f{\"u}r die Entwicklung mechanistischer tierversuchsfreier Ans{\"a}tze: Eine Fallstudie {\"u}ber Nephrotoxizit{\"a}t initiiert durch rezeptorvermittelte Endozytose und lysosomalen Overload},
  doi       = {10.25972/OPUS-31804},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-318046},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2023},
  abstract  = {Zur Verbesserung der Pr{\"u}fung und Risikobewertung der zunehmenden Menge von Chemikalien und Arzneimitteln, gilt es neue Alternativen in Form von in vitro Pr{\"u}fmethoden mit mechanistisch relevanten Endpunkten zu finden. Einen solchen Rahmen bietet das konzeptionelle Konstrukt des Adverse Outcome Pathway (AOP)- Konzepts. Es erzeugt auf der Basis bestehenden Wissens einen mechanistischen und kausalen Zusammenhang mit Hilfe von mehreren Schl{\"u}sselereignissen (Key Event [KE]) zwischen einem initierenden molekularen Ereignis (Molecular Initiating Event [MIE]) und einem adversen Effekt (Adverse Outcome [AO]) auf biologischer Ebene. Im Rahmen dieser Arbeit wurde der AOP „Rezeptorvermittelte Endozytose und lysosomaler Overload f{\"u}hren zu Nephrotoxizit{\"a}t" am Zellkulturmodell proximaler Nierentubuluszellen weiterentwickelt. Es wurden in vitro Assays f{\"u}r die Zelllinien RPTEC/TERT1 (Mensch) und NRK-52 E (Ratte) f{\"u}r jedes KE etabliert. In dem AOP wird die Initiierung der Sch{\"a}digung des Nierengewebes durch rezeptorvermittelte Endozytose der Substanzen (MIE) mit folgendem lysosomalem Overload (KE 1) und der lysosomalen Membranruptur (KE 2) beschrieben. Es kommt zur Zellsch{\"a}digung (KE 3) und endet mit einem Schaden auf Organebene (AO). F{\"u}r KE 1 erfolgte die Visualisierung des lysosomal-assoziierten Membranproteins (lysosomal-associated Membranprotein [LAMP]) und in KE 2 die Darstellung der Protease Cathepsin D (CTSD) mittels Immunfluoreszenz. F{\"u}r KE 3 wurden spezifische Toxizit{\"a}tsdaten der Testsubstanzen mit dem CellTiter-Glo® Lumineszenz-Zellviabilit{\"a}tstest generiert. Gew{\"a}hlte Stressoren f{\"u}r den AOP war die Gruppe der Polymyxin-Antibiotika (Polymyxin B, Colistin, Polymyxin B Nonapeptid), das Aminoglykosid Gentamicin, das Glykopeptid Vancomycin sowie Cadmiumchlorid. In Zusammenschau der Ergebnisse der drei KEs war die Rangfolge der Auswirkungen der drei Polymyxin-Derivate {\"u}ber alle KEs konsistent. Polymyxin B erwies sich als aktivste Substanz, w{\"a}hrend Polymyxin B Nonapeptid die geringsten Auswirkungen zeigte. Als Ausblick in weiterf{\"u}hrenden Analysen der Arbeitsgruppe konnten bei Cadmiumchlorid trotz einer signifikanten Zytotoxizit{\"a}t (KE 3) nur geringe Auswirkungen in der LAMPExpression (KE 1) aufgezeigt werden. Des Weiteren erfolgte die Erstellung von Response-Response-Analysen, um mittels vorgeschalteter Schl{\"u}sselereignisse nachfolgende Effekte vorhersagen zu k{\"o}nnen. Projektpartner der Universit{\"a}t Utrecht entwickelten dar{\"u}ber hinaus eine quantitative in vitro in vivo Extrapolation (QIVIVE) mittels eines physiologisch basierten pharmakokinetischen (PBPK) Modells.},
  subject      = {Nephrotoxizit{\"a}t},
  language  = {de}
}
@article{PaisdziorDimitriouSchoepeetal.2020,
  author    = {Paisdzior, Sarah and Dimitriou, Ioanna Maria and Sch{\"o}pe, Paul Curtis and Annibale, Paolo and Scheerer, Patrick and Krude, Heiko and Lohse, Martin J. and Biebermann, Heike and K{\"u}hnen, Peter},
  title     = {Differential signaling profiles of MC4R mutations with three different ligands},
  series = {International Journal of Molecular Sciences},
  volume    = {21},
  journal   = {International Journal of Molecular Sciences},
  number    = {4},
  issn      = {1422-0067},
  doi       = {10.3390/ijms21041224},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-285108},
  year      = {2020},
  abstract  = {The melanocortin 4 receptor (MC4R) is a key player in hypothalamic weight regulation and energy expenditure as part of the leptin-melanocortin pathway. Mutations in this G protein coupled receptor (GPCR) are the most common cause for monogenetic obesity, which appears to be mediated by changes in the anorectic action of MC4R via G\(_S\)-dependent cyclic adenosine-monophosphate (cAMP) signaling as well as other signaling pathways. To study potential bias in the effects of MC4R mutations between the different signaling pathways, we investigated three major MC4R mutations: a G\(_S\) loss-of-function (S127L) and a G\(_S\) gain-of-function mutant (H158R), as well as the most common European single nucleotide polymorphism (V103I). We tested signaling of all four major G protein families plus extracellular regulated kinase (ERK) phosphorylation and β-arrestin2 recruitment, using the two endogenous agonists, α- and β-melanocyte stimulating hormone (MSH), along with a synthetic peptide agonist (NDP-α-MSH). The S127L mutation led to a full loss-of-function in all investigated pathways, whereas V103I and H158R were clearly biased towards the G\(_{q/11}\) pathway when challenged with the endogenous ligands. These results show that MC4R mutations can cause vastly different changes in the various MC4R signaling pathways and highlight the importance of a comprehensive characterization of receptor mutations.},
  language  = {en}
}
@article{MaurerHartmannArgyriouetal.2022,
  author    = {Maurer, Wiebke and Hartmann, Nico and Argyriou, Loukas and Sossalla, Samuel and Streckfuss-B{\"o}meke, Katrin},
  title     = {Generation of homozygous Na\(_{v}\)1.8 knock-out iPSC lines by CRISPR Cas9 genome editing to investigate a potential new antiarrhythmic strategy},
  series = {Stem Cell Research},
  volume    = {60},
  journal   = {Stem Cell Research},
  doi       = {10.1016/j.scr.2022.102677},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-300936},
  year      = {2022},
  abstract  = {The sodium channel Na\(_{v}\)1.8, encoded by SCN10A, is reported to contribute to arrhythmogenesis by inducing the late I\(_{Na}\) and thereby enhanced persistent Na\(^{+}\) current. However, its exact electrophysiological role in cardiomyocytes remains unclear. Here, we generated induced pluripotent stem cells (iPSCs) with a homozygous SCN10A knock-out from a healthy iPSC line by CRISPR Cas9 genome editing. The edited iPSCs maintained full pluripotency, genomic integrity, and spontaneous in vitro differentiation capacity. The iPSCs are able to differentiate into iPSC-cardiomyocytes, hence making it possible to investigate the role of Na\(_{v}\)1.8 in the heart.},
  language  = {en}
}
@phdthesis{Eppli2023,
  author    = {Eppli, Nenad},
  title     = {Untersuchung des Einflusses der ERK1/2-Autophosphorylierung an Threonin 188 auf Mausherzen mittels transgener M{\"a}use mit ubiquit{\"a}rer {\"U}berexpression von ERK2\(^{T188D}\)},
  doi       = {10.25972/OPUS-21655},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-216558},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2023},
  abstract  = {Die ERK2Thr188-Autophosphoylierung stellt einen regulatorischen Signalweg dar, der infolge einer hypertrophen Stimulation die kardiale Hypertrophie beg{\"u}nstigt. Eine Hemmung dieser Phosphorylierung in Kardiomyozyten verhindert die Ausbildung der kardialen Hypertrophie ohne Beeinflussung der kardioprotektiven Funktionen von ERK1/2. Demgegen{\"u}ber f{\"u}hrt die dauerhafte Simulation zu einem gain-of-function-Ph{\"a}notypen mit ausgepr{\"a}gter Hypertophie, Fibrose und einer reduzierten Herzfunktion. In dieser Arbeit wurde die dauerhafte Simulation ERK2Thr188-Phosphorylierung (T188D) in einem Mausmodell mit ubiquit{\"a}rer Expression dieser Mutation untersucht. Dabei konnte gezeigt werden, dass sich nach Stimulation durch TAC in diesen Tieren ein etwas st{\"a}rkerer hypertropher Ph{\"a}notyp mit vergr{\"o}ßerten Kardiomyozyten, gesteigerter interstitieller Fibrosierung und reduzierter Herzfunktion ausbildet als in M{\"a}usen mit kardiomyozyten-spezifischer {\"U}berexpression diese Mutante. In Fibroblasten- und VSMC-Zelllinien wurde eine gesteigerte Proliferation der T188D-{\"u}berexprimierenden Zellen im Vergleich zu Kontrollen festgestellt. Somit scheint die ERK2Thr188-Phosphorylierung auch in kardialen Nicht-Myozyten einen maladaptiven Einfluss auf das Herz auszu{\"u}ben.},
  subject      = {Herzhypertrophie},
  language  = {de}
}
@phdthesis{Zink2023,
  author    = {Zink, Christoph},
  title     = {Biochemische und strukturbiologische Charakterisierung der Inhibition der Pyridoxal 5´-Phosphat Phosphatase durch 7,8-Dihydroxyflavon},
  doi       = {10.25972/OPUS-25151},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-251511},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2023},
  abstract  = {Die Pyridoxal-5'-Phosphat Phosphatase (PDXP), auch bekannt als Chronophin (CIN), ist eine HAD-Phosphatase, die beim Menschen ubiquit{\"a}r exprimiert wird und eine entscheidende Rolle im zellul{\"a}ren Vitamin-B6-Metabolismus einnimmt. PDXP ist in der Lage Pyridoxal-5'-Phosphat (PLP), die co-enzymatisch aktive Form von Vitamin B6, zu dephosphorylieren. In-vivo Studien mit M{\"a}usen zeigten, dass die Abwesenheit von PDXP mit verbesserten kognitiven Leistungen und einem verringerten Wachstum von Hirntumoren assoziiert ist. Dies begr{\"u}ndet die gezielte Suche nach einem pharmakologischen Inhibitor f{\"u}r PDXP. Ein Hochdurchsatz-Screen legte nahe, dass 7,8-Dihydroxyflavon (7,8-DHF) hierf{\"u}r ein potenzieller Kandidat ist. Zahlreiche Studien beschreiben bereits vielf{\"a}ltige positive neurologische Effekte nach in-vivo Administration von 7,8-DHF, allerdings bleibt der genaue Wirkmechanismus umstritten und wird bis dato nicht mit PDXP in Zusammenhang gebracht. Ziel dieser Arbeit ist es, die Inhibition von PDXP durch 7,8-DHF n{\"a}her zu charakterisieren und damit einen Beitrag zur Beantwortung der Frage zu leisten, ob PDXP an den 7,8-DHF-induzierten Effekten beteiligt ist. Hierzu wurde der Effekt von 7,8-DHF auf die enzymatische Aktivit{\"a}t von rekombinant hergestelltem, gereinigtem PDXP in in-vitro Phosphatase-Assays charakterisiert. Um die Selektivit{\"a}t von 7,8-DHF gegen{\"u}ber PDXP zu untersuchen, wurden f{\"u}nf weitere HAD-Phosphatasen getestet. Unter den analysierten Phosphatasen zeigte einzig die dem PDXP nah verwandte Phosphoglykolat Phosphatase (PGP) eine geringer ausgepr{\"a}gte Sensitivit{\"a}t gegen 7,8-DHF. Ein Vergleich von 7,8-DHF mit sechs strukturell verwandten, hydroxylierten Flavonen zeigte, dass 7,8-DHF unter den getesteten Substanzen die h{\"o}chste Potenz und Effektivit{\"a}t aufwies. Außerdem wurde eine Co-Kristallisation von PDXP mit 7,8-DHF durchgef{\"u}hrt, deren Struktur bis zu einer Aufl{\"o}sung von 2,0 {\AA} verfeinert werden konnte. Die in der Kristallstruktur identifizierte Bindungsstelle von 7,8-DHF an PDXP wurde mittels verschiedener, neu generierter PDXP-Mutanten enzymkinetisch best{\"a}tigt. Zusammenfassend zeigen die hier beschriebenen Ergebnisse, dass 7,8-DHF ein direkter, selektiver und vorwiegend kompetitiver Inhibitor der PDXP-Aktivit{\"a}t ist, mit einer IC50 im submikromolaren Bereich. Die Ergebnisse dieser in-vitro Untersuchungen motivieren zu weiterer Forschung bez{\"u}glich der 7,8-DHF-vermittelten Inhibition der PDXP-Aktivit{\"a}t in Zellen, um die Frage beantworten zu k{\"o}nnen, ob PDXP auch in-vivo ein relevantes Target f{\"u}r 7,8-DHF darstellt.},
  subject      = {Pyridoxalphosphat},
  language  = {de}
}
@article{NwoghaAbtewRaveendranetal.2023,
  author    = {Nwogha, Jeremiah S. and Abtew, Wosene G. and Raveendran, Muthurajan and Oselebe, Happiness O. and Obidiegwu, Jude E. and Chilaka, Cynthia A. and Amirtham, Damodarasamy D.},
  title     = {Role of non-structural sugar metabolism in regulating tuber dormancy in white yam (Dioscorea rotundata)},
  series = {Agriculture},
  volume    = {13},
  journal   = {Agriculture},
  number    = {2},
  issn      = {2077-0472},
  doi       = {10.3390/agriculture13020343},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-304486},
  year      = {2023},
  abstract  = {Changes in sugar composition occur continuously in plant tissues at different developmental stages. Tuber dormancy induction, stability, and breaking are very critical developmental transitions in yam crop production. Prolonged tuber dormancy after physiological maturity has constituted a great challenge in yam genetic improvement and productivity. In the present study, biochemical profiling of non-structural sugar in yam tubers during dormancy was performed to determine the role of non-structural sugar in yam tuber dormancy regulation. Two genotypes of the white yam species, one local genotype (Obiaoturugo) and one improved genotype (TDr1100873), were used for this study. Tubers were sampled at 42, 56, 87, 101, 115, and 143 days after physiological maturity (DAPM). Obiaoturugo exhibited a short dormant phenotype and sprouted at 101-DAPM, whereas TDr1100873 exhibited a long dormant phenotype and sprouted at 143-DAPM. Significant metabolic changes were observed in non-structural sugar parameters, dry matter, and moisture content in Obiaoturugo from 56-DAPM, whereas in TDr1100873, significant metabolic changes were observed from 101-DAPM. It was observed that the onset of these metabolic changes occurred at a point when the tubers of both genotypes exhibited a dry matter content of 60\%, indicating that a dry matter content of 60\% might be a critical threshold for white yam tuber sprouting. Non-reducing sugars increased by 9-10-fold during sprouting in both genotypes, which indicates their key role in tuber dormancy regulation in white yam. This result implicates that some key sugar metabolites can be targeted for dormancy manipulation of the yam crop.},
  language  = {en}
}
@article{HadiBankogluStopper2023,
  author    = {Hadi, Naji Said Aboud and Bankoglu, Ezgi Eyluel and Stopper, Helga},
  title     = {Genotoxicity of pyrrolizidine alkaloids in metabolically inactive human cervical cancer HeLa cells co-cultured with human hepatoma HepG2 cells},
  series = {Archives of Toxicology},
  volume    = {97},
  journal   = {Archives of Toxicology},
  number    = {1},
  doi       = {10.1007/s00204-022-03394-z},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-324708},
  pages     = {295-306},
  year      = {2023},
  abstract  = {Pyrrolizidine alkaloids (PAs) are secondary plant metabolites, which can be found as contaminant in various foods and herbal products. Several PAs can cause hepatotoxicity and liver cancer via damaging hepatic sinusoidal endothelial cells (HSECs) after hepatic metabolization. HSECs themselves do not express the required metabolic enzymes for activation of PAs. Here we applied a co-culture model to mimic the in vivo hepatic environment and to study PA-induced effects on not metabolically active neighbour cells. In this co-culture model, bioactivation of PA was enabled by metabolically capable human hepatoma cells HepG2, which excrete the toxic and mutagenic pyrrole metabolites. The human cervical epithelial HeLa cells tagged with H2B-GFP were utilized as non-metabolically active neighbours because they can be identified easily based on their green fluorescence in the co-culture. The PAs europine, riddelliine and lasiocarpine induced micronuclei in HepG2 cells, and in HeLa H2B-GFP cells co-cultured with HepG2 cells, but not in HeLa H2B-GFP cells cultured alone. Metabolic inhibition of cytochrome P450 enzymes with ketoconazole abrogated micronucleus formation. The efflux transporter inhibitors verapamil and benzbromarone reduced micronucleus formation in the co-culture model. Furthermore, mitotic disturbances as an additional genotoxic mechanism of action were observed in HepG2 cells and in HeLa H2B-GFP cells co-cultured with HepG2 cells, but not in HeLa H2B-GFP cells cultured alone. Overall, we were able to show that PAs were activated by HepG2 cells and the metabolites induced genomic damage in co-cultured HeLa cells.},
  language  = {en}
}
@article{BittnerBoonDelbancoetal.2022,
  author    = {Bittner, Nataly and Boon, Andy and Delbanco, Evert H. and Walter, Christof and Mally, Angela},
  title     = {Assessment of aromatic amides in printed food contact materials: analysis of potential cleavage to primary aromatic amines during simulated passage through the gastrointestinal tract},
  series = {Archives of Toxicology},
  volume    = {96},
  journal   = {Archives of Toxicology},
  number    = {5},
  doi       = {10.1007/s00204-022-03254-w},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-324697},
  pages     = {1423-1435},
  year      = {2022},
  abstract  = {Recent analyses conducted by German official food control reported detection of the aromatic amides N-(2,4-dimethylphenyl)acetamide (NDPA), N-acetoacetyl-m-xylidine (NAAX) and 3-hydroxy-2-naphthanilide (Naphthol AS) in cold water extracts from certain food contact materials made from paper or cardboard, including paper straws, paper napkins, and cupcake liners. Because aromatic amides may be cleaved to potentially genotoxic primary amines upon oral intake, these findings raise concern that transfer of NDPA, NAAX and Naphthol AS from food contact materials into food may present a risk to human health. The aim of the present work was to assess the stability of NDPA, NAAX and Naphthol AS and potential cleavage to 2,4-dimethylaniline (2,4-DMA) and aniline during simulated passage through the gastrointestinal tract using static in vitro digestion models. Using the digestion model established by the National Institute for Public Health and the Environment (RIVM, Bilthoven, NL) and a protocol recommended by the European Food Safety Authority, potential hydrolysis of the aromatic amides to the respective aromatic amines was assessed by LC-MS/MS following incubation of the aromatic amides with digestive fluid simulants. Time-dependent hydrolysis of NDPA and NAAX resulting in formation of the primary aromatic amine 2,4-DMA was consistently observed in both models. The highest rate of cleavage of NDPA and NAAX was recorded following 4 h incubation with 0.07 M HCl as gastric-juice simulant, and amounted to 0.21\% and 0.053\%, respectively. Incubation of Naphthol AS with digestive fluid simulants did not give rise to an increase in the concentration of aniline above the background that resulted from the presence of aniline as an impurity of the test compound. Considering the lack of evidence for aniline formation from Naphthol AS and the extremely low rate of hydrolysis of the amide bonds of NDPA and NAAX during simulated passage through the gastrointestinal tract that gives rise to only very minor amounts of the potentially mutagenic and/or carcinogenic aromatic amine 2,4-DMA, risk assessment based on assumption of 100\% cleavage to the primary aromatic amines would appear to overestimate health risks related to the presence of aromatic amides in food contact materials.},
  language  = {en}
}
@article{NaseemOthmanFathyetal.2020,
  author    = {Naseem, Muhammad and Othman, Eman M. and Fathy, Moustafa and Iqbal, Jibran and Howari, Fares M. and AlRemeithi, Fatima A. and Kodandaraman, Geema and Stopper, Helga and Bencurova, Elena and Vlachakis, Dimitrios and Dandekar, Thomas},
  title     = {Integrated structural and functional analysis of the protective effects of kinetin against oxidative stress in mammalian cellular systems},
  series = {Scientific Reports},
  volume    = {10},
  journal   = {Scientific Reports},
  doi       = {10.1038/s41598-020-70253-1},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-231317},
  year      = {2020},
  abstract  = {Metabolism and signaling of cytokinins was first established in plants, followed by cytokinin discoveries in all kingdoms of life. However, understanding of their role in mammalian cells is still scarce. Kinetin is a cytokinin that mitigates the effects of oxidative stress in mammalian cells. The effective concentrations of exogenously applied kinetin in invoking various cellular responses are not well standardized. Likewise, the metabolism of kinetin and its cellular targets within the mammalian cells are still not well studied. Applying vitality tests as well as comet assays under normal and hyper-oxidative states, our analysis suggests that kinetin concentrations of 500 nM and above cause cytotoxicity as well as genotoxicity in various cell types. However, concentrations below 100 nM do not cause any toxicity, rather in this range kinetin counteracts oxidative burst and cytotoxicity. We focus here on these effects. To get insights into the cellular targets of kinetin mediating these pro-survival functions and protective effects we applied structural and computational approaches on two previously testified targets for these effects. Our analysis deciphers vital residues in adenine phosphoribosyltransferase (APRT) and adenosine receptor (A2A-R) that facilitate the binding of kinetin to these two important human cellular proteins. We finally discuss how the therapeutic potential of kinetin against oxidative stress helps in various pathophysiological conditions.},
  language  = {en}
}
@article{MaiellaroLohseKitteetal.2016,
  author    = {Maiellaro, Isabella and Lohse, Martin J. and Kitte, Robert J. and Calebiro, Davide},
  title     = {cAMP Signals in Drosophila Motor Neurons Are Confined to Single Synaptic Boutons},
  series = {Cell Reports},
  volume    = {17},
  journal   = {Cell Reports},
  number    = {5},
  doi       = {10.1016/j.celrep.2016.09.090},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-162324},
  pages     = {1238-1246},
  year      = {2016},
  abstract  = {The second messenger cyclic AMP (cAMP) plays an important role in synaptic plasticity. Although there is evidence for local control of synaptic transmission and plasticity, it is less clear whether a similar spatial confinement of cAMP signaling exists. Here, we suggest a possible biophysical basis for the site-specific regulation of synaptic plasticity by cAMP, a highly diffusible small molecule that transforms the physiology of synapses in a local and specific manner. By exploiting the octopaminergic system of Drosophila, which mediates structural synaptic plasticity via a cAMP-dependent pathway, we demonstrate the existence of local cAMP signaling compartments of micrometer dimensions within single motor neurons. In addition, we provide evidence that heterogeneous octopamine receptor localization, coupled with local differences in phosphodiesterase activity, underlies the observed differences in cAMP signaling in the axon, cell body, and boutons.},
  language  = {en}
}
@article{WeigandRonchiVanselowetal.2021,
  author    = {Weigand, Isabel and Ronchi, Cristina L. and Vanselow, Jens T. and Bathon, Kerstin and Lenz, Kerstin and Herterich, Sabine and Schlosser, Andreas and Kroiss, Matthias and Fassnacht, Martin and Calebiro, Davide and Sbiera, Silviu},
  title     = {PKA Cα subunit mutation triggers caspase-dependent RIIβ subunit degradation via Ser\(^{114}\) phosphorylation},
  series = {Science Advances},
  volume    = {7},
  journal   = {Science Advances},
  number    = {8},
  doi       = {10.1126/sciadv.abd4176},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-270445},
  year      = {2021},
  abstract  = {Mutations in the PRKACA gene are the most frequent cause of cortisol-producing adrenocortical adenomas leading to Cushing's syndrome. PRKACA encodes for the catalytic subunit α of protein kinase A (PKA). We already showed that PRKACA mutations lead to impairment of regulatory (R) subunit binding. Furthermore, PRKACA mutations are associated with reduced RIIβ protein levels; however, the mechanisms leading to reduced RIIβ levels are presently unknown. Here, we investigate the effects of the most frequent PRKACA mutation, L206R, on regulatory subunit stability. We find that Ser\(^{114}\) phosphorylation of RIIβ is required for its degradation, mediated by caspase 16. Last, we show that the resulting reduction in RIIβ protein levels leads to increased cortisol secretion in adrenocortical cells. These findings reveal the molecular mechanisms and pathophysiological relevance of the R subunit degradation caused by PRKACA mutations, adding another dimension to the deregulation of PKA signaling caused by PRKACA mutations in adrenal Cushing's syndrome.},
  language  = {en}
}
@article{AwadOthmanStopper2017,
  author    = {Awad, Eman and Othman, Eman M. and Stopper, Helga},
  title     = {Effects of resveratrol, lovastatin and the mTOR-inhibitor RAD-001 on insulin-induced genomic damage in vitro},
  series = {Molecules},
  volume    = {22},
  journal   = {Molecules},
  number    = {12},
  doi       = {10.3390/molecules22122207},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-159260},
  pages     = {2207},
  year      = {2017},
  abstract  = {Diabetes mellitus (DM) is one of the major current health problems due to lifestyle changes. Before diagnosis and in the early years of disease, insulin blood levels are elevated. However, insulin generates low levels of reactive oxygen species (ROS) which are integral to the regulation of a variety of intracellular signaling pathways, but excess levels of insulin may also lead to DNA oxidation and DNA damage. Three pharmaceutical compounds, resveratrol, lovastatin and the mTOR-inhibitor RAD-001, were investigated due to their known beneficial effects. They showed protective properties against genotoxic damage and significantly reduced ROS after in vitro treatment of cultured cells with insulin. Therefore, the selected pharmaceuticals may be attractive candidates to be considered for support of DM therapy.},
  language  = {en}
}
@article{LohseBockMaiellaroetal.2017,
  author    = {Lohse, Christian and Bock, Andreas and Maiellaro, Isabella and Hannawacker, Annette and Schad, Lothar R. and Lohse, Martin J. and Bauer, Wolfgang R.},
  title     = {Experimental and mathematical analysis of cAMP nanodomains},
  series = {PLoS ONE},
  volume    = {12},
  journal   = {PLoS ONE},
  number    = {4},
  doi       = {10.1371/journal.pone.0174856},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-170972},
  pages     = {e0174856},
  year      = {2017},
  abstract  = {In their role as second messengers, cyclic nucleotides such as cAMP have a variety of intracellular effects. These complex tasks demand a highly organized orchestration of spatially and temporally confined cAMP action which should be best achieved by compartmentalization of the latter. A great body of evidence suggests that cAMP compartments may be established and maintained by cAMP degrading enzymes, e.g. phosphodiesterases (PDEs). However, the molecular and biophysical details of how PDEs can orchestrate cAMP gradients are entirely unclear. In this paper, using fusion proteins of cAMP FRET-sensors and PDEs in living cells, we provide direct experimental evidence that the cAMP concentration in the vicinity of an individual PDE molecule is below the detection limit of our FRET sensors (<100nM). This cAMP gradient persists in crude cytosol preparations. We developed mathematical models based on diffusion-reaction equations which describe the creation of nanocompartments around a single PDE molecule and more complex spatial PDE arrangements. The analytically solvable equations derived here explicitly determine how the capability of a single PDE, or PDE complexes, to create a nanocompartment depend on the cAMP degradation rate, the diffusive mobility of cAMP, and geometrical and topological parameters. We apply these generic models to our experimental data and determine the diffusive mobility and degradation rate of cAMP. The results obtained for these parameters differ by far from data in literature for free soluble cAMP interacting with PDE. Hence, restricted cAMP diffusion in the vincinity of PDE is necessary to create cAMP nanocompartments in cells.},
  language  = {en}
}
@article{SchuppStopperHeidland2016,
  author    = {Schupp, Nicole and Stopper, Helga and Heidland, August},
  title     = {DNA Damage in Chronic Kidney Disease: Evaluation of Clinical Biomarkers},
  series = {Oxidative Medicine and Cellular Longevity},
  volume    = {2016},
  journal   = {Oxidative Medicine and Cellular Longevity},
  number    = {3592042},
  doi       = {10.1155/2016/3592042},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-166569},
  year      = {2016},
  abstract  = {Patients with chronic kidney disease (CKD) exhibit an increased cancer risk compared to a healthy control population. To be able to estimate the cancer risk of the patients and to assess the impact of interventional therapies thereon, it is of particular interest to measure the patients' burden of genomic damage. Chromosomal abnormalities, reduced DNA repair, and DNA lesions were found indeed in cells of patients with CKD. Biomarkers for DNA damage measurable in easily accessible cells like peripheral blood lymphocytes are chromosomal aberrations, structural DNA lesions, and oxidatively modified DNA bases. In this review the most common methods quantifying the three parameters mentioned above, the cytokinesis-block micronucleus assay, the comet assay, and the quantification of 8-oxo-7,8-dihydro-2′-deoxyguanosine, are evaluated concerning the feasibility of the analysis and regarding the marker's potential to predict clinical outcomes.},
  language  = {en}
}
@article{FathyFawzyHintzscheetal.2019,
  author    = {Fathy, Moustafa and Fawzy, Michael Atef and Hintzsche, Henning and Nikaido, Toshio and Dandekar, Thomas and Othman, Eman M.},
  title     = {Eugenol exerts apoptotic effect and modulates the sensitivity of HeLa cells to cisplatin and radiation},
  series = {Molecules},
  volume    = {24},
  journal   = {Molecules},
  number    = {21},
  issn      = {1420-3049},
  doi       = {10.3390/molecules24213979},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-193227},
  pages     = {3979},
  year      = {2019},
  abstract  = {Eugenol is a phytochemical present in different plant products, e.g., clove oil. Traditionally, it is used against a number of different disorders and it was suggested to have anticancer activity. In this study, the activity of eugenol was evaluated in a human cervical cancer (HeLa) cell line and cell proliferation was examined after treatment with various concentrations of eugenol and different treatment durations. Cytotoxicity was tested using lactate dehydrogenase (LDH) enzyme leakage. In order to assess eugenol's potential to act synergistically with chemotherapy and radiotherapy, cell survival was calculated after eugenol treatment in combination with cisplatin and X-rays. To elucidate its mechanism of action, caspase-3 activity was analyzed and the expression of various genes and proteins was checked by RT-PCR and western blot analyses. Eugenol clearly decreased the proliferation rate and increased LDH release in a concentration- and time-dependent manner. It showed synergistic effects with cisplatin and X-rays. Eugenol increased caspase-3 activity and the expression of Bax, cytochrome c (Cyt-c), caspase-3, and caspase-9 and decreased the expression of B-cell lymphoma (Bcl)-2, cyclooxygenase-2 (Cox-2), and interleukin-1 beta (IL-1β) indicating that eugenol mainly induced cell death by apoptosis. In conclusion, eugenol showed antiproliferative and cytotoxic effects via apoptosis and also synergism with cisplatin and ionizing radiation in the human cervical cancer cell line.},
  language  = {en}
}
@article{WoelfelSaetteleZechmeisteretal.2020,
  author    = {W{\"o}lfel, Angela and S{\"a}ttele, Mathias and Zechmeister, Christina and Nikolaev, Viacheslov O. and Lohse, Martin J. and Boege, Fritz and Jahns, Roland and Boivin-Jahns, Val{\´e}rie},
  title     = {Unmasking features of the auto-epitope essential for β\(_1\)-adrenoceptor activation by autoantibodies in chronic heart failure},
  series = {ESC Heart Failure},
  volume    = {7},
  journal   = {ESC Heart Failure},
  number    = {4},
  doi       = {10.1002/ehf2.12747},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-235974},
  pages     = {1830-1841},
  year      = {2020},
  abstract  = {Aims Chronic heart failure (CHF) can be caused by autoantibodies stimulating the heart via binding to first and/or second extracellular loops of cardiac β1-adrenoceptors. Allosteric receptor activation depends on conformational features of the autoantibody binding site. Elucidating these features will pave the way for the development of specific diagnostics and therapeutics. Our aim was (i) to fine-map the conformational epitope within the second extracellular loop of the human β\(_1\)-adrenoceptor (β1ECII) that is targeted by stimulating β\(_1\)-receptor (auto)antibodies and (ii) to generate competitive cyclopeptide inhibitors of allosteric receptor activation, which faithfully conserve the conformational auto-epitope. Methods and results Non-conserved amino acids within the β\(_1\)EC\(_{II}\) loop (compared with the amino acids constituting the ECII loop of the β\(_2\)-adrenoceptor) were one by one replaced with alanine; potential intra-loop disulfide bridges were probed by cysteine-serine exchanges. Effects on antibody binding and allosteric receptor activation were assessed (i) by (auto)antibody neutralization using cyclopeptides mimicking β1ECII ± the above replacements, and (ii) by (auto)antibody stimulation of human β\(_1\)-adrenoceptors bearing corresponding point mutations. With the use of stimulating β\(_1\)-receptor (auto)antibodies raised in mice, rats, or rabbits and isolated from exemplary dilated cardiomyopathy patients, our series of experiments unmasked two features of the β\(_1\)EC\(_{II}\) loop essential for (auto)antibody binding and allosteric receptor activation: (i) the NDPK\(^{211-214}\) motif and (ii) the intra-loop disulfide bond C\(^{209}\)↔C\(^{215}\). Of note, aberrant intra-loop disulfide bond C\(^{209}\)↔C\(^{216}\) almost fully disrupted the functional auto-epitope in cyclopeptides. Conclusions The conformational auto-epitope targeted by cardio-pathogenic β\(_1\)-receptor autoantibodies is faithfully conserved in cyclopeptide homologues of the β\(_1\)EC\(_{II}\) loop bearing the NDPK\(^{211-214}\) motif and the C\(^{209}\)↔C\(^{215}\) bridge while lacking cysteine C216. Such molecules provide promising tools for novel diagnostic and therapeutic approaches in β\(_1\)-autoantibodypositive CHF.},
  language  = {en}
}
@article{ScholzGuanNieberleretal.2017,
  author    = {Scholz, Nicole and Guan, Chonglin and Nieberler, Matthias and Grotmeyer, Alexander and Maiellaro, Isabella and Gao, Shiqiang and Beck, Sebastian and Pawlak, Matthias and Sauer, Markus and Asan, Esther and Rothemund, Sven and Winkler, Jana and Pr{\"o}mel, Simone and Nagel, Georg and Langenhan, Tobias and Kittel, Robert J},
  title     = {Mechano-dependent signaling by Latrophilin/CIRL quenches cAMP in proprioceptive neurons},
  series = {eLife},
  volume    = {6},
  journal   = {eLife},
  number    = {e28360},
  doi       = {10.7554/eLife.28360},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-170520},
  year      = {2017},
  abstract  = {Adhesion-type G protein-coupled receptors (aGPCRs), a large molecule family with over 30 members in humans, operate in organ development, brain function and govern immunological responses. Correspondingly, this receptor family is linked to a multitude of diverse human diseases. aGPCRs have been suggested to possess mechanosensory properties, though their mechanism of action is fully unknown. Here we show that the Drosophila aGPCR Latrophilin/dCIRL acts in mechanosensory neurons by modulating ionotropic receptor currents, the initiating step of cellular mechanosensation. This process depends on the length of the extended ectodomain and the tethered agonist of the receptor, but not on its autoproteolysis, a characteristic biochemical feature of the aGPCR family. Intracellularly, dCIRL quenches cAMP levels upon mechanical activation thereby specifically increasing the mechanosensitivity of neurons. These results provide direct evidence that the aGPCR dCIRL acts as a molecular sensor and signal transducer that detects and converts mechanical stimuli into a metabotropic response.},
  language  = {en}
}
@article{GmachBathePetersTeluguetal.2022,
  author    = {Gmach, Philipp and Bathe-Peters, Marc and Telugu, Narasimha and Miller, Duncan C. and Annibale, Paolo},
  title     = {Fluorescence spectroscopy of low-level endogenous β-adrenergic receptor expression at the plasma membrane of differentiating human iPSC-derived cardiomyocytes},
  series = {International Journal of Molecular Sciences},
  volume    = {23},
  journal   = {International Journal of Molecular Sciences},
  number    = {18},
  issn      = {1422-0067},
  doi       = {10.3390/ijms231810405},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-288277},
  year      = {2022},
  abstract  = {The potential of human-induced pluripotent stem cells (hiPSCs) to be differentiated into cardiomyocytes (CMs) mimicking adult CMs functional morphology, marker genes and signaling characteristics has been investigated since over a decade. The evolution of the membrane localization of CM-specific G protein-coupled receptors throughout differentiation has received, however, only limited attention to date. We employ here advanced fluorescent spectroscopy, namely linescan Fluorescence Correlation Spectroscopy (FCS), to observe how the plasma membrane abundance of the β\(_1\)- and β\(_2\)-adrenergic receptors (β\(_{1/2}\)-ARs), labelled using a bright and photostable fluorescent antagonist, evolves during the long-term monolayer culture of hiPSC-derived CMs. We compare it to the kinetics of observed mRNA levels in wildtype (WT) hiPSCs and in two CRISPR/Cas9 knock-in clones. We conduct these observations against the backdrop of our recent report of cell-to-cell expression variability, as well as of the subcellular localization heterogeneity of β-ARs in adult CMs.},
  language  = {en}
}
@article{SchanbacherHermannsLorenzetal.2023,
  author    = {Schanbacher, Constanze and Hermanns, Heike M. and Lorenz, Kristina and Wajant, Harald and Lang, Isabell},
  title     = {Complement 1q/tumor necrosis factor-related proteins (CTRPs): structure, receptors and signaling},
  series = {Biomedicines},
  volume    = {11},
  journal   = {Biomedicines},
  number    = {2},
  issn      = {2227-9059},
  doi       = {10.3390/biomedicines11020559},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-304136},
  year      = {2023},
  abstract  = {Adiponectin and the other 15 members of the complement 1q (C1q)/tumor necrosis factor (TNF)-related protein (CTRP) family are secreted proteins composed of an N-terminal variable domain followed by a stalk region and a characteristic C-terminal trimerizing globular C1q (gC1q) domain originally identified in the subunits of the complement protein C1q. We performed a basic PubMed literature search for articles mentioning the various CTRPs or their receptors in the abstract or title. In this narrative review, we briefly summarize the biology of CTRPs and focus then on the structure, receptors and major signaling pathways of CTRPs. Analyses of CTRP knockout mice and CTRP transgenic mice gave overwhelming evidence for the relevance of the anti-inflammatory and insulin-sensitizing effects of CTRPs in autoimmune diseases, obesity, atherosclerosis and cardiac dysfunction. CTRPs form homo- and heterotypic trimers and oligomers which can have different activities. The receptors of some CTRPs are unknown and some receptors are redundantly targeted by several CTRPs. The way in which CTRPs activate their receptors to trigger downstream signaling pathways is largely unknown. CTRPs and their receptors are considered as promising therapeutic targets but their translational usage is still hampered by the limited knowledge of CTRP redundancy and CTRP signal transduction.},
  language  = {en}
}
@article{JanzWalzCirnuetal.2024,
  author    = {Janz, Anna and Walz, Katharina and Cirnu, Alexandra and Surjanto, Jessica and Urlaub, Daniela and Leskien, Miriam and Kohlhaas, Michael and Nickel, Alexander and Brand, Theresa and Nose, Naoko and W{\"o}rsd{\"o}rfer, Philipp and Wagner, Nicole and Higuchi, Takahiro and Maack, Christoph and Dudek, Jan and Lorenz, Kristina and Klopocki, Eva and Erg{\"u}n, S{\"u}leyman and Duff, Henry J. and Gerull, Brenda},
  title     = {Mutations in DNAJC19 cause altered mitochondrial structure and increased mitochondrial respiration in human iPSC-derived cardiomyocytes},
  series = {Molecular Metabolism},
  volume    = {79},
  journal   = {Molecular Metabolism},
  issn      = {2212-8778},
  doi       = {10.1016/j.molmet.2023.101859},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-350393},
  year      = {2024},
  abstract  = {Highlights • Loss of DNAJC19's DnaJ domain disrupts cardiac mitochondrial structure, leading to abnormal cristae formation in iPSC-CMs. • Impaired mitochondrial structures lead to an increased mitochondrial respiration, ROS and an elevated membrane potential. • Mutant iPSC-CMs show sarcomere dysfunction and a trend to more arrhythmias, resembling DCMA-associated cardiomyopathy. Background Dilated cardiomyopathy with ataxia (DCMA) is an autosomal recessive disorder arising from truncating mutations in DNAJC19, which encodes an inner mitochondrial membrane protein. Clinical features include an early onset, often life-threatening, cardiomyopathy associated with other metabolic features. Here, we aim to understand the metabolic and pathophysiological mechanisms of mutant DNAJC19 for the development of cardiomyopathy. Methods We generated induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) of two affected siblings with DCMA and a gene-edited truncation variant (tv) of DNAJC19 which all lack the conserved DnaJ interaction domain. The mutant iPSC-CMs and their respective control cells were subjected to various analyses, including assessments of morphology, metabolic function, and physiological consequences such as Ca\(^{2+}\) kinetics, contractility, and arrhythmic potential. Validation of respiration analysis was done in a gene-edited HeLa cell line (DNAJC19tv\(_{HeLa}\)). Results Structural analyses revealed mitochondrial fragmentation and abnormal cristae formation associated with an overall reduced mitochondrial protein expression in mutant iPSC-CMs. Morphological alterations were associated with higher oxygen consumption rates (OCRs) in all three mutant iPSC-CMs, indicating higher electron transport chain activity to meet cellular ATP demands. Additionally, increased extracellular acidification rates suggested an increase in overall metabolic flux, while radioactive tracer uptake studies revealed decreased fatty acid uptake and utilization of glucose. Mutant iPSC-CMs also showed increased reactive oxygen species (ROS) and an elevated mitochondrial membrane potential. Increased mitochondrial respiration with pyruvate and malate as substrates was observed in mutant DNAJC19tv HeLa cells in addition to an upregulation of respiratory chain complexes, while cellular ATP-levels remain the same. Moreover, mitochondrial alterations were associated with increased beating frequencies, elevated diastolic Ca\(^{2+}\) concentrations, reduced sarcomere shortening and an increased beat-to-beat rate variability in mutant cell lines in response to β-adrenergic stimulation. Conclusions Loss of the DnaJ domain disturbs cardiac mitochondrial structure with abnormal cristae formation and leads to mitochondrial dysfunction, suggesting that DNAJC19 plays an essential role in mitochondrial morphogenesis and biogenesis. Moreover, increased mitochondrial respiration, altered substrate utilization, increased ROS production and abnormal Ca\(^{2+}\) kinetics provide insights into the pathogenesis of DCMA-related cardiomyopathy.},
  language  = {en}
}
@article{GuthHueserRothetal.2021,
  author    = {Guth, Sabine and H{\"u}ser, Stephanie and Roth, Angelika and Degen, Gisela and Diel, Patrick and Edlund, Karolina and Eisenbrand, Gerhard and Engel, Karl-Heinz and Epe, Bernd and Grune, Tilman and Heinz, Volker and Henle, Thomas and Humpf, Hans-Ulrich and J{\"a}ger, Henry and Joost, Hans-Georg and Kulling, Sabine E. and Lampen, Alfonso and Mally, Angela and Marchan, Rosemarie and Marko, Doris and M{\"u}hle, Eva and Nitsche, Michael A. and R{\"o}hrdanz, Elke and Stadler, Richard and van Thriel, Christoph and Vieths, Stefan and Vogel, Rudi F. and Wascher, Edmund and Watzl, Carsten and N{\"o}thlings, Ute and Hengstler, Jan G.},
  title     = {Contribution to the ongoing discussion on fluoride toxicity},
  series = {Archives of Toxicology},
  volume    = {95},
  journal   = {Archives of Toxicology},
  number    = {7},
  issn      = {0340-5761},
  doi       = {10.1007/s00204-021-03072-6},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-307161},
  pages     = {2571-2587},
  year      = {2021},
  abstract  = {Since the addition of fluoride to drinking water in the 1940s, there have been frequent and sometimes heated discussions regarding its benefits and risks. In a recently published review, we addressed the question if current exposure levels in Europe represent a risk to human health. This review was discussed in an editorial asking why we did not calculate benchmark doses (BMD) of fluoride neurotoxicity for humans. Here, we address the question, why it is problematic to calculate BMDs based on the currently available data. Briefly, the conclusions of the available studies are not homogeneous, reporting negative as well as positive results; moreover, the positive studies lack control of confounding factors such as the influence of well-known neurotoxicants. We also discuss the limitations of several further epidemiological studies that did not meet the inclusion criteria of our review. Finally, it is important to not only focus on epidemiological studies. Rather, risk analysis should consider all available data, including epidemiological, animal, as well as in vitro studies. Despite remaining uncertainties, the totality of evidence does not support the notion that fluoride should be considered a human developmental neurotoxicant at current exposure levels in European countries.},
  language  = {en}
}
@article{BarileBerryBlaauboeretal.2021,
  author    = {Barile, Frank A. and Berry, Colin and Blaauboer, Bas and Boobis, Alan and Bolt, Herrmann M. and Borgert, Christopher and Dekant, Wolfgang and Dietrich, Daniel and Domingo, Jose L. and Galli, Corrado L. and Gori, Gio Batta and Greim, Helmut and Hengstler, Jan G. and Heslop-Harrison, Pat and Kacew, Sam and Marquardt, Hans and Mally, Angela and Pelkonen, Olavi and Savolainen, Kai and Testai, Emanuela and Tsatsakis, Aristides and Vermeulen, Nico P.},
  title     = {The EU chemicals strategy for sustainability: in support of the BfR position},
  series = {Archives of Toxicology},
  volume    = {95},
  journal   = {Archives of Toxicology},
  number    = {9},
  issn      = {0340-5761},
  doi       = {10.1007/s00204-021-03125-w},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-307154},
  pages     = {3133-3136},
  year      = {2021},
  abstract  = {The EU chemicals strategy for sustainability (CSS) asserts that both human health and the environment are presently threatened and that further regulation is necessary. In a recent Guest Editorial, members of the German competent authority for risk assessment, the BfR, raised concerns about the scientific justification for this strategy. The complexity and interdependence of the networks of regulation of chemical substances have ensured that public health and wellbeing in the EU have continuously improved. A continuous process of improvement in consumer protection is clearly desirable but any initiative directed towards this objective must be based on scientific knowledge. It must not confound risk with other factors in determining policy. This conclusion is fully supported in the present Commentary including the request to improve both, data collection and the time-consuming and bureaucratic procedures that delay the publication of regulations.},
  language  = {en}
}
@article{RebsStreckfussBoemeke2023,
  author    = {Rebs, Sabine and Streckfuss-B{\"o}meke, Katrin},
  title     = {How can we use stem cell-derived cardiomyocytes to understand the involvement of energetic metabolism in alterations of cardiac function?},
  series = {Frontiers in Molecular Medicine},
  volume    = {3},
  journal   = {Frontiers in Molecular Medicine},
  doi       = {10.3389/fmmed.2023.1222986},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-327344},
  year      = {2023},
  abstract  = {Mutations in the mitochondrial-DNA or mitochondria related nuclear-encoded-DNA lead to various multisystemic disorders collectively termed mitochondrial diseases. One in three cases of mitochondrial disease affects the heart muscle, which is called mitochondrial cardiomyopathy (MCM) and is associated with hypertrophic, dilated, and noncompact cardiomyopathy. The heart is an organ with high energy demand, and mitochondria occupy 30\%-40\% of its cardiomyocyte-cell volume. Mitochondrial dysfunction leads to energy depletion and has detrimental effects on cardiac performance. However, disease development and progression in the context of mitochondrial and nuclear DNA mutations, remains incompletely understood. The system of induced pluripotent stem cell (iPSC)-derived cardiomyocytes (CM) is an excellent platform to study MCM since the unique genetic identity to their donors enables a robust recapitulation of the predicted phenotypes in a dish on a patient-specific level. Here, we focus on recent insights into MCM studied by patient-specific iPSC-CM and further discuss research gaps and advances in metabolic maturation of iPSC-CM, which is crucial for the study of mitochondrial dysfunction and to develop novel therapeutic strategies.},
  language  = {en}
}
@unpublished{BrennerZinkWitzingeretal.2024,
  author    = {Brenner, Marian and Zink, Christoph and Witzinger, Linda and Keller, Angelika and Hadamek, Kerstin and Bothe, Sebastian and Neuenschwander, Martin and Villmann, Carmen and von Kries, Jens Peter and Schindelin, Hermann and Jeanclos, Elisabeth and Gohla, Antje},
  title     = {7,8-Dihydroxyflavone is a direct inhibitor of pyridoxal phosphatase},
  series = {eLife},
  journal   = {eLife},
  doi       = {10.7554/eLife.93094.2},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-350446},
  year      = {2024},
  abstract  = {Vitamin B6 deficiency has been linked to cognitive impairment in human brain disorders for decades. Still, the molecular mechanisms linking vitamin B6 to these pathologies remain poorly understood, and whether vitamin B6 supplementation improves cognition is unclear as well. Pyridoxal phosphatase (PDXP), an enzyme that controls levels of pyridoxal 5'-phosphate (PLP), the co-enzymatically active form of vitamin B6, may represent an alternative therapeutic entry point into vitamin B6-associated pathologies. However, pharmacological PDXP inhibitors to test this concept are lacking. We now identify a PDXP and age-dependent decline of PLP levels in the murine hippocampus that provides a rationale for the development of PDXP inhibitors. Using a combination of small molecule screening, protein crystallography and biolayer interferometry, we discover and analyze 7,8-dihydroxyflavone (7,8-DHF) as a direct and potent PDXP inhibitor. 7,8-DHF binds and reversibly inhibits PDXP with low micromolar affinity and sub-micromolar potency. In mouse hippocampal neurons, 7,8-DHF increases PLP in a PDXP-dependent manner. These findings validate PDXP as a druggable target. Of note, 7,8-DHF is a well-studied molecule in brain disorder models, although its mechanism of action is actively debated. Our discovery of 7,8-DHF as a PDXP inhibitor offers novel mechanistic insights into the controversy surrounding 7,8-DHF-mediated effects in the brain.},
  language  = {en}
}
@article{DjelićBorozanDimitrijevićSrećkovićetal.2022,
  author    = {Djelić, Ninoslav and Borozan, Sunčica and Dimitrijević-Srećković, Vesna and Pajović, Nevena and Mirilović, Milorad and Stopper, Helga and Stanimirović, Zoran},
  title     = {Oxidative stress and DNA damage in peripheral blood mononuclear cells from normal, obese, prediabetic and diabetic persons exposed to thyroid hormone in vitro},
  series = {International Journal of Molecular Sciences},
  volume    = {23},
  journal   = {International Journal of Molecular Sciences},
  number    = {16},
  issn      = {1422-0067},
  doi       = {10.3390/ijms23169072},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-285988},
  year      = {2022},
  abstract  = {Diabetes, a chronic group of medical disorders characterized byhyperglycemia, has become a global pandemic. Some hormones may influence the course and outcome of diabetes, especially if they potentiate the formation of reactive oxygen species (ROS). There is a close relationship between thyroid disorders and diabetes. The main objective of this investigation was to find out whether peripheral blood mononuclear cells (PBMCs) are more prone to DNA damage by triiodothyronine (T\(_3\)) (0.1, 1 and 10 μM) at various stages of progression through diabetes (obese, prediabetics, and type 2 diabetes mellitus—T2DM persons). In addition, some biochemical parameters of oxidative stress (catalase-CAT, thiobarbituric acid reactive substances—TBARS) and lactate dehydrogenase (LDH) were evaluated. PBMCs from prediabetic and diabetic patients exhibited increased sensitivity for T\(_3\) regarding elevated level of DNA damage, inhibition of catalase, and increase of TBARS and LDH. PBMCs from obese patients reacted in the same manner, except for DNA damage. The results of this study should contribute to a better understanding of the role of thyroid hormones in the progression of T2DM.},
  language  = {en}
}
@phdthesis{Horn2024,
  author    = {Horn, Daniela},
  title     = {Kardiotoxizit{\"a}t von CTRPs und das Vorkommen der CTRP-Rezeptoren in Kardiomyozyten},
  doi       = {10.25972/OPUS-34902},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-349029},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2024},
  abstract  = {Die C1q/tumor necrosis factor-related proteins (CTRPs) sind eine Ligandenfamilie aus sezernierten Plasmaproteinen, welche sich in ihrem Grundbauplan {\"a}hneln. Daten aus der Literatur deuten darauf hin, dass sie zum Teil positive Effekte auf den Stoffwechsel und das Herz-Kreislaufsystem besitzen und somit eine m{\"o}gliche therapeutische Zielstruktur darstellen. W{\"a}hrend f{\"u}r manche CTRPs bereits Rezeptoren identifiziert werden konnten, ist f{\"u}r andere immer noch nicht gekl{\"a}rt, an welche Rezeptoren sie binden oder {\"u}ber welche sie diese Wirkungen erzielen. Um die CTRPs zuk{\"u}nftig therapeutisch nutzen zu k{\"o}nnen, muss die Wirkung der CTRPs auf verschiedene Zellen weiter analysiert werden. Daf{\"u}r wurden in dieser Arbeit Zellen, auf die Expression bereits bekannter CTRP-Rezeptoren hin, untersucht. Des Weiteren wurden die durch CTRP2, CTRP3, CTRP4, CTRP9A, CTRP10, CTRP11, CTRP13 und CTRP14 induzierten {\"A}nderungen in der ATP- und Laktatproduktion als Surrogatparameter f{\"u}r Kardiotoxizit{\"a}t in den Kardiomyozytenzelllinien H9c2 und AC16 getestet, um potenziell kardiotoxische Wirkungen fr{\"u}hzeitig erkennen zu k{\"o}nnen. Es konnte gezeigt werden, dass die CTRPs sicher f{\"u}r Kardiomyozyten zu sein scheinen, was eine wichtige Grundlage f{\"u}r die therapeutische Nutzbarkeit darstellt.},
  subject      = {Herzmuskelzelle},
  language  = {de}
}
@article{EberlRebsHoppeetal.2024,
  author    = {Eberl, Hanna and Rebs, Sabine and Hoppe, Stefanie and Sedaghat-Hamedani, Farbod and Kayvanpour, Elham and Meder, Benjamin and Streckfuss-B{\"o}meke, Katrin},
  title     = {Generation of an RBM20-mutation-associated left-ventricular non-compaction cardiomyopathy iPSC line (UMGi255-A) into a DCM genetic background to investigate monogenetic cardiomyopathies},
  series = {Stem Cell Research},
  volume    = {74},
  journal   = {Stem Cell Research},
  issn      = {1873-5061},
  doi       = {10.1016/j.scr.2023.103290},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-350565},
  year      = {2024},
  abstract  = {RBM20 mutations account for 3 \% of genetic cardiomypathies and manifest with high penetrance and arrhythmogenic effects. Numerous mutations in the conserved RS domain have been described as causing dilated cardiomyopathy (DCM), whereas a particular mutation (p.R634L) drives development of a different cardiac phenotype: left-ventricular non-compaction cardiomyopathy. We generated a mutation-induced pluripotent stem cell (iPSC) line in which the RBM20-LVNC mutation p.R634L was introduced into a DCM patient line with rescued RBM20-p.R634W mutation. These DCM-634L-iPSC can be differentiated into functional cardiomyocytes to test whether this RBM20 mutation induces development of the LVNC phenotype within the genetic context of a DCM patient.},
  language  = {en}
}
@article{BankogluStippGerberetal.2021,
  author    = {Bankoglu, Ezgi Eyluel and Stipp, Franzisca and Gerber, Johanna and Seyfried, Florian and Heidland, August and Bahner, Udo and Stopper, Helga},
  title     = {Effect of cryopreservation on DNA damage and DNA repair activity in human blood samples in the comet assay},
  series = {Archives of Toxicology},
  volume    = {95},
  journal   = {Archives of Toxicology},
  number    = {5},
  doi       = {10.1007/s00204-021-03012-4},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-265326},
  pages     = {1831-1841},
  year      = {2021},
  abstract  = {The comet assay is a commonly used method to determine DNA damage and repair activity in many types of samples. In recent years, the use of the comet assay in human biomonitoring became highly attractive due to its various modified versions, which may be useful to determine individual susceptibility in blood samples. However, in human biomonitoring studies, working with large sample numbers that are acquired over an extended time period requires some additional considerations. One of the most important issues is the storage of samples and its effect on the outcome of the comet assay. Another important question is the suitability of different blood preparations. In this study, we analysed the effect of cryopreservation on DNA damage and repair activity in human blood samples. In addition, we investigated the suitability of different blood preparations. The alkaline and FPG as well as two different types of repair comet assay and an in vitro hydrogen peroxide challenge were applied. Our results confirmed that cryopreserved blood preparations are suitable for investigating DNA damage in the alkaline and FPG comet assay in whole blood, buffy coat and PBMCs. Ex vivo hydrogen peroxide challenge yielded its optimal effect in isolated PBMCs. The utilised repair comet assay with either UVC or hydrogen peroxide-induced lesions and an aphidicolin block worked well in fresh PBMCs. Cryopreserved PBMCs could not be used immediately after thawing. However, a 16-h recovery with or without mitotic stimulation enabled the application of the repair comet assay, albeit only in a surviving cell fraction.},
  language  = {en}
}
@article{ChristianSeierDrakopoulosetal.2020,
  author    = {Christian, Gentzsch and Seier, Kerstin and Drakopoulos, Antonios and Jobin, Marie-Lise and Lanoisel{\´e}e, Yann and Koszegi, Zsombor and Maurel, Damien and Sounier, R{\´e}my and H{\"u}bner, Harald and Gmeiner, Peter and Granier, S{\´e}bastien and Calebiro, Davide and Decker, Michael},
  title     = {Selective and Wash-Resistant Fluorescent Dihydrocodeinone Derivatives Allow Single-Molecule Imaging of μ-Opioid Receptor Dimerization},
  series = {Angewandte Chemie International Edition},
  volume    = {59},
  journal   = {Angewandte Chemie International Edition},
  number    = {15},
  doi       = {10.1002/anie.201912683},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-212398},
  pages     = {5958-5964},
  year      = {2020},
  abstract  = {μ-Opioid receptors (μ-ORs) play a critical role in the modulation of pain and mediate the effects of the most powerful analgesic drugs. Despite extensive efforts, it remains insufficiently understood how μ-ORs produce specific effects in living cells. We developed new fluorescent ligands based on the μ-OR antagonist E-p-nitrocinnamoylamino-dihydrocodeinone (CACO), that display high affinity, long residence time and pronounced selectivity. Using these ligands, we achieved single-molecule imaging of μ-ORs on the surface of living cells at physiological expression levels. Our results reveal a high heterogeneity in the diffusion of μ-ORs, with a relevant immobile fraction. Using a pair of fluorescent ligands of different color, we provide evidence that μ-ORs interact with each other to form short-lived homodimers on the plasma membrane. This approach provides a new strategy to investigate μ-OR pharmacology at single-molecule level.},
  language  = {en}
}
@article{GodboleLygaLohseetal.2017,
  author    = {Godbole, Amod and Lyga, Sandra and Lohse, Martin J. and Calebiro, Davide},
  title     = {Internalized TSH receptors en route to the TGN induce local G\(_{S}\)-protein signaling and gene transcription},
  series = {Nature Communications},
  volume    = {8},
  journal   = {Nature Communications},
  number    = {443},
  doi       = {10.1038/s41467-017-00357-2},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-170375},
  year      = {2017},
  abstract  = {A new paradigm of G-protein-coupled receptor (GPCR) signaling at intracellular sites has recently emerged, but the underlying mechanisms and functional consequences are insufficiently understood. Here, we show that upon internalization in thyroid cells, endogenous TSH receptors traffic retrogradely to the trans-Golgi network (TGN) and activate endogenous Gs-proteins in the retromer-coated compartment that brings them to the TGN. Receptor internalization is associated with a late cAMP/protein kinase A (PKA) response at the Golgi/TGN. Blocking receptor internalization, inhibiting PKA II/interfering with its Golgi/TGN localization, silencing retromer or disrupting Golgi/TGN organization all impair efficient TSH-dependent cAMP response element binding protein (CREB) phosphorylation. These results suggest that retrograde trafficking to the TGN induces local G\(_{S}\)-protein activation and cAMP/PKA signaling at a critical position near the nucleus, which appears required for efficient CREB phosphorylation and gene transcription. This provides a new mechanism to explain the functional consequences of GPCR signaling at intracellular sites and reveals a critical role for the TGN in GPCR signaling.},
  language  = {en}
}
@article{MaurerHuppBischoffetal.2017,
  author    = {Maurer, Jana and Hupp, Sabrina and Bischoff, Carolin and Foertsch, Christina and Mitchell, Timothy J. and Chakraborty, Trinad and Iliev, Asparouh I.},
  title     = {Distinct neurotoxicity profile of listeriolysin O from \(Listeria\) \(monocytogenes\)},
  series = {Toxins},
  volume    = {9},
  journal   = {Toxins},
  number    = {1},
  doi       = {10.3390/toxins9010034},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-172130},
  year      = {2017},
  abstract  = {Cholesterol-dependent cytolysins (CDCs) are protein toxins that originate from Gram-positive bacteria and contribute substantially to their pathogenicity. CDCs bind membrane cholesterol and build prepores and lytic pores. Some effects of the toxins are observed in non-lytic concentrations. Two pathogens, \(Streptococcus\) \(pneumoniae\) and \(Listeria\) \(monocytogenes\), cause fatal bacterial meningitis, and both produce toxins of the CDC family—pneumolysin and listeriolysin O, respectively. It has been demonstrated that pneumolysin produces dendritic varicosities (dendrite swellings) and dendritic spine collapse in the mouse neocortex, followed by synaptic loss and astrocyte cell shape remodeling without elevated cell death. We utilized primary glial cultures and acute mouse brain slices to examine the neuropathological effects of listeriolysin O and to compare it to pneumolysin with identical hemolytic activity. In cultures, listeriolysin O permeabilized cells slower than pneumolysin did but still initiated non-lytic astrocytic cell shape changes, just as pneumolysin did. In an acute brain slice culture system, listeriolysin O produced dendritic varicosities in an NMDA-dependent manner but failed to cause dendritic spine collapse and cortical astrocyte reorganization. Thus, listeriolysin O demonstrated slower cell permeabilization and milder glial cell remodeling ability than did pneumolysin and lacked dendritic spine collapse capacity but exhibited equivalent dendritic pathology.},
  language  = {en}
}
@phdthesis{Cirkel2018,
  author    = {Cirkel, Nanett Christin},
  title     = {Beeinflussung des oxidativen Stress-Status in einem Rattenmodell: Effekt von Selenmangel auf Niere und Leber},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-163631},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2018},
  abstract  = {Das Spurenelement Selen und Vitamin E reduzieren reaktive Sauerstoff Spezies (ROS). Bei Mangel dieser wichtigen Stoffe erh{\"o}ht sich die Konzentration an ROS und der oxidative Stress steigt. Unter erh{\"o}hten ROS entstehen vermehrt DNA-Sch{\"a}den und Lipidperoxidationen. Das ROS Wasserstoffperoxid wird zu Wasser {\"u}ber das Enzym Gluthationperxoidase reduziert. Dessen Aktivit{\"a}t steigert Selen um den Faktor 100-1.000. Das Aktivit{\"a}tsmaximum des Enzyms liegt bei einer t{\"a}glichen Selenaufnahme von 60-80 Mikrogramm/Tag. Dadurch wird die Menge an ROS reduziert und der oxidative Stress in der Zelle nimmt ab. Vitamin E fungiert als Radikalf{\"a}nger. Sein Derivat alpha- Tocopherol besitzt die h{\"o}chste antioxidative Wirkung und kann Lipidperoxidationen unterbrechen. Die vorliegende Arbeit untersucht Auswirkungen von oxidativem Stress, den ein Mangel von Selen und Vitamin E in der Nahrung bei 6 Monate und 12 Monate alten Tieren auf Leber und Niere verursacht. Der Nachweis von oxidativem Stress erfolgte {\"u}ber sogenannte Hitzeschockproteine HSP70 und H{\"a}moxygenase 1. HSP 70 wird auch unter physiologischen Bedingungen exprimiert. Es wirkt als Chaperon und ist u.a. f{\"u}r die korrekte Faltung und Stabilisierung von Proteinen zust{\"a}ndig. Die Versuche zeigten, dass im Alter in der Niere die HSP70 Konzentration ansteigt und die Zelle unter vermehrtem oxidativen Stress leidet. Entsprechende Literaturergebnisse wurden best{\"a}tigt. Die H{\"a}moxygenase 1 (HO-1) ist ein Schl{\"u}sselenzym, das vermehrt bei oxidativem Stress gebildet wird. Hoch reaktionsfreudige und freie Blutbestandteile katalysiert die H{\"a}moxygenase. Einen Abfall der HO- 1 Konzentration zeigten Untersuchungen von Leber und Niere bei Selen, - Vitamin E Mangel und h{\"o}herem Lebensalter. Gr{\"u}nde f{\"u}r die verminderte Expression sind noch wenig erforscht. Die vermehrte Anreicherung von Superoxidanionradikalen wurde in den Geweben von Leber und Niere {\"u}ber Dihydroethidium (DHE) F{\"a}rbung nachgewiesen. Die Hypothese wurde best{\"a}tigt, dass bei Selen, -Vitamin E Mangelnahrung und h{\"o}herem Alter vermehrter oxidativer Stress entsteht. Selenmangel beg{\"u}nstigt die Entstehung verschiedener Krankheiten, z.B. Krebs, koronale Herzerkrankung und vor allem die Keshan-Krankheit, die den Herzmuskel bef{\"a}llt. Selen nimmt positiven Einfluss auf K{\"o}rperfunktionen: Fertilit{\"a}t, embryonalen Entwicklung und Entwicklung eines Neugeborenen. Einige Fragen bleiben ungekl{\"a}rt: Welche physiologischen Entwicklungsprozesse f{\"o}rdert Selen? Nimmt Selen eine wichtige Funktion bei der Befruchtung der Eizelle ein? Wie beeinflusst Selen die Entwicklung des Gehirns? Dem Spurenelement Selen kommen offensichtlich neben seiner Bedeutung zur Minderung des oxidativen Stresses noch weitere wichtige Funktionen zu, die bisher wenig untersucht wurden.},
  subject      = {Oxidativer Stress},
  language  = {de}
}
@phdthesis{Soliman2022,
  author    = {Soliman, Alexander},
  title     = {Einfluss des Gewichtsverlusts auf den oxidativen Stress und den DNS-Schaden in adip{\"o}sen Patient*innen nach bariatrischer Chirurgie},
  doi       = {10.25972/OPUS-27835},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-278354},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2022},
  abstract  = {Adipositas ist eine Erkrankung, die durch ein erh{\"o}htes Krebsrisiko neben zahlreichen anderen Komorbidit{\"a}ten mit weitreichenden Folgen f{\"u}r die Gesundheit adip{\"o}ser Patient*innen einhergeht. In der Pathogenese der adipositas-assoziierten Krebsarten sind dabei ein erh{\"o}hter oxidativer Stress sowie die damit einhergehende Sch{\"a}digung der DNS maßgeblich beteiligt. Im Umkehrschluss wurde in der vorliegenden Arbeit der Einfluss eines durch bariatrische Chirurgie induzierten Gewichtsverlusts auf den oxidativen Stress und DNS-Schaden in adip{\"o}sen Patient*innen anhand von Blutproben pr{\"a}operativ sowie 6 und 12 Monate postoperativ untersucht. In einer Subpopulation der Patient*innen konnte eine tendenzielle Verringerung des DNS-Schadens anhand des Comet-Assays in peripheren Lymphozyten beobachtet werden. Im Hinblick auf den oxidativen Stress wurde im Plasma die Eisenreduktionsf{\"a}higkeit als Maß f{\"u}r die antioxidative Kapazit{\"a}t sowie Malondialdehyd als Surrogatmarker f{\"u}r das Ausmaß an Lipidperoxidation bestimmt. Weiterhin wurde in Erythrozyten das Gesamtglutathion und das oxidierte Glutathion bestimmt. Die oxidativen Stressparameter zeigten insgesamt nach einer initialen Zunahme im oxidativen Stress 6 Monate postoperativ eine r{\"u}ckl{\"a}ufige Tendenz im oxidativen Stress am Studienende. Somit geben die Beobachtungen dieser Arbeit Anlass zur Hoffnung, dass adip{\"o}se Patient*innen durch einen bariatrisch induzierten Gewichtsverlust von einer Verringerung des Krebsrisikos profitieren k{\"o}nnten.},
  subject      = {Magenchirurgie},
  language  = {de}
}
@article{BauerMallyLiedtke2021,
  author    = {Bauer, Benedikt and Mally, Angela and Liedtke, Daniel},
  title     = {Zebrafish embryos and larvae as alternative animal models for toxicity testing},
  series = {International Journal of Molecular Sciences},
  volume    = {22},
  journal   = {International Journal of Molecular Sciences},
  number    = {24},
  issn      = {1422-0067},
  doi       = {10.3390/ijms222413417},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-284225},
  year      = {2021},
  abstract  = {Prerequisite to any biological laboratory assay employing living animals is consideration about its necessity, feasibility, ethics and the potential harm caused during an experiment. The imperative of these thoughts has led to the formulation of the 3R-principle, which today is a pivotal scientific standard of animal experimentation worldwide. The rising amount of laboratory investigations utilizing living animals throughout the last decades, either for regulatory concerns or for basic science, demands the development of alternative methods in accordance with 3R to help reduce experiments in mammals. This demand has resulted in investigation of additional vertebrate species displaying favourable biological properties. One prominent species among these is the zebrafish (Danio rerio), as these small laboratory ray-finned fish are well established in science today and feature outstanding biological characteristics. In this review, we highlight the advantages and general prerequisites of zebrafish embryos and larvae before free-feeding stages for toxicological testing, with a particular focus on cardio-, neuro, hepato- and nephrotoxicity. Furthermore, we discuss toxicokinetics, current advances in utilizing zebrafish for organ toxicity testing and highlight how advanced laboratory methods (such as automation, advanced imaging and genetic techniques) can refine future toxicological studies in this species.},
  language  = {en}
}