Originalpublikationen und Übersichtsartikel

2017

Abdulmughni A, Jozwik IK, Putkaradze N, Brill E, Zapp J, Thunnissen AW, Hannemann F, Bernhardt R.(2017) Characterization of cytochrome P450 CYP109E1 from Bacillus megaterium as a novel vitamin D3 hydroxylase. J Biotechnol. 243:38-47.

Putkaradze N, Kiss FM, Schmitz D, Zapp J, Hutter MC, Bernhardt R.(2017) Biotransformation of prednisone and dexamethasone by cytochrome P450 based systems - Identification of new potential drug candidates. J Biotechnol. 242:101-110.

2016

Litzenburger M, Lo Izzo R, Bernhardt R, Khatri Y.(2016) Investigating the roles of T224 and T232 in the oxidation of cinnamaldehyde catalyzed by myxobacterial CYP260B1. FEBS Lett. doi: 10.1002/1873-3468.12519

Khatri Y, Carius Y, Ringle M, Lancaster CR, Bernhardt R.(2016) Structural characterization of CYP260A1 from Sorangium cellulosum to investigate the 1α-hydroxylation of a mineralocorticoid. FEBS Lett. doi: 10.1002/1873-3468.12479

Milhim M, Putkaradze N, Abdulmughni A, Kern F, Hartz P, Bernhardt R.(2016) Identification of a new plasmid-encoded cytochrome P450 CYP107DY1 from Bacillus megaterium with a catalytic activity towards mevastatin. J Biotechnol. 240:68-75.

Jozwik IK, Kiss FM, Gricman L, Abdulmughni A, Brill E, Zapp J, Pleiss J, Bernhardt R, Thunnissen AW.(2016) Structural basis of steroid binding and oxidation by the cytochrome P450 CYP109E1 from Bacillus megaterium. FEBS J. 283(22): 4128-4148.

Milhim M, Gerber A, Neunzig J, Hannemann F, Bernhardt R.(2016) A Novel NADPH-dependent flavoprotein reductase from Bacillus megaterium acts as an efficient cytochrome P450 reductase. J Biotechnol. 231:83-94.

Gerber A, Milhim M, Hartz P, Zapp J, Bernhardt R.(2016) Genetic engineering of Bacillus megaterium for high-yield production of the major teleost progestogens 17α,20β-di- and 17α,20β,21α-trihydroxy-4-pregnen-3-one. Metab Eng. 36:19-27.

Salamanca-Pinzon SG, Khatri Y, Carius Y, Keller L, Müller R, Lancaster CR, Bernhardt R. (2016) Structure-function analysis for the hydroxylation of 4 C21-steroids by the myxobacterial CYP260B1. FEBS Lett. 590(12):1838-51.

Schiffer L, Müller AR, Hobler A, Brixius-Anderko S, Zapp J, Hannemann F, Bernhardt R. (2016) Biotransformation of the mineralocorticoid receptor antagonists spironolactone and canrenone by human CYP11B1 and CYP11B2: Characterization of the products and their influence on mineralocorticoid receptor transactivation. J Steroid Biochem Mol Biol. 163:68-76.

Khatri Y, Ringle M, Lisurek M, von Kries JP, Zapp J, Bernhardt R. (2016) Substrate Hunting for the Myxobacterial CYP260A1 Revealed New 1α-Hydroxylated Products from C-19 Steroids. Chembiochem. 1:90-101.

Mosa A, Gerber A, Neunzig J, Bernhardt R. (2016) Products of gut-microbial tryptophan metabolism inhibit the steroid hormone-synthesizing cytochrome P450 11A1. Endocrine. 53(2):610-4.

Litzenburger M, Bernhardt R. (2016) Selective oxidation of carotenoid-derived aroma compounds by CYP260B1 and CYP267B1 from Sorangium cellulosum So ce56. Appl Microbiol Biotechnol. 100(10):4447-57.

Bernhardt R. (2016) The potential of targeting CYP11B. Expert Opin Ther Targets. 20(8):923-34.

Schifrin A, Khatri Y, Kirsch P, Thiel V, Schulz S, Bernhardt R. (2016) A single terpene synthase is responsible for a wide variety of sesquiterpenes in Sorangium cellulosum Soce56. Org Biomo Chem. 14(13):3385-93.

Brixius-Anderko S, Hannemann F, Ringle M, Khatri Y, Bernhardt R. (2016) An indole deficient Escherichia coli strain improves screening of cytochromes P450 for biotechnological applications. Biotechnol Appl Biochem. doi: 10.1002/bab.1488.

Janocha S, Carius Y, Hutter M, Lancaster CR, Bernhardt R. (2016) Crystal Structure of CYP106A2 in Substrate-Free and Substrate-Bound Form. Chembiochem. 17(9):852-60.

Ehrhardt M, Gerber A, Zapp J, Hannemann F, Bernhardt R. (2016) Human CYP27A1 catalyzes hydroxylation of β-sitosterol and ergosterol. Biol Chem. 397(6):513-8.

Kern F, Khatri Y, Litzenburger M, Bernhardt R. (2016) CYP267A1 and CYP267B1 from Sorangium cellulosum So ce56 are Highly Versatile Drug Metabolizers. Drug Metab Dispos. 44(4):495-504.

Schiffer L, Brixius-Anderko S, Hannemann F, Zapp J, Neunzig J, Thevis M, Bernhardt R. (2016) Metabolism of Oral Turinabol by Human Steroid Hormone-Synthesizing Cytochrome P450 Enzymes. Chembiochem. 44(2):227-37.

Ehrhardt M, Gerber A, Hannemann F, Bernhardt R. (2016) Expression of human CYP27A1 in B. megaterium for the efficient hydroxylation of cholesterol, vitamin D3 and 7-dehydrocholesterol. J Biotechnol. 218:34-40.

Nguyen HH, Eiden-Plach A, Hannemann F, Malunowicz EM, Hartmann MF, Wudy SA, Bernhardt R. (2016) Phenotypic, metabolic, and molecular genetic characterization of six patients with congenital adrenal hyperplasia caused by novel mutations in the CYP11B1 gene. J Steroid Biochem Mol Biol. 155:126-34.

2015

Kern F, Dier TK, Khatri Y, Ewen KM, Jacquot JP, Volmer DA, Bernhardt R. (2015) Highly Efficient CYP167A1 (EpoK) dependent Epothilone B Formation and Production of 7-Ketone Epothilone D as a New Epothilone Derivative. sci Rep. 1:8;5:14881.

Kiss FM., Khatri Y., Zapp J., Bernhardt R. (2015) Identification of new substrates for the CYP106A1-mediated 11-oxidation and investigation of the reaction mechanism. FEBS Lett. 589:2320-2326.

Litzenburger M., Kern F., Khatri Y., Bernhardt R. (2015) Conversions of tricyclic antidepressants and antipsychotics with selected P450s from Sorangium cellulosum So ce56. Drug Metab. Dispos. 43:392-399.

Mosa A., Hutter MC., Zapp J., Bernhardt R., Hannemann F.(2015) Regioselective Acetylation of C21 Hydroxysteroids by the Bacterial Chloramphenicol Acetyltransferase I. Chembiochem. 16:1670-1679.

Gerber A., Kleser M., Biedendieck R., Bernhardt R., Hannemann F. (2015) Functionalized PHB granules provide the basis for the efficient side-chain cleavage of cholesterol and analogs in recombinant Bacillus megaterium. Microb Cell Fact. 14:107

Schiffer L., Anderko S., Hobler A., Hannemann F., Kagawa N., Bernhardt R. (2015) A recombinant CYP11B1 dependent Escherichia coli biocatalyst for selective cortisol production and optimization towards a preparative scale. Microb Cell Fact. 14:25.

Kiss FM., Schmitz D., Zapp J., Dier TK., Volmer DA., Bernhardt R. (2015) Comparison of CYP106A1 and CYP106A2 from Bacillus megaterium identification of a novel 11-oxidase activity. Appl Microbiol Biotechnol. 24.

Mosa A., Neunzig J., Gerber A., Zapp J., Hannemann F., Pilak P., Bernhardt R. (2015) 2beta- and 16beta-hydroxylase activity of CYP11A1 and direct stimulatory effect of estrogens on pregnenolone formation. J Steroid Biochem Mol Biol. 150:-1-10.

Schifrin A., Ly TT., Günnewich N., Zapp J., Thiel V., Schulz S., Hannemann F., Khatri Y., Bernhardt R. (2015) Characterization of the gene cluster CYP264B1-geoA from Sorangium cellulosum So ce56: biosynthesis of (+)-eremophilene and its hydroxylation. Chembiochem. 16(2):337-344.

Schiffer L., Anderko S., Hannemann F. ,Eiden-Plach A., Bernhardt R. (2015). The CYP11B subfamily (Review). J Steroid Biochem Mol Biol. 151:38-51.

Khatri Y., Hannemann F., Girhard M., Kappl R., Hutter M., Urlacher VB., Bernhardt R.(2015) A natural heme-signature variant of CYP267A1 from Sorangium cellulosum So ce56 executes diverse ω-hydroxylation. FEBS J. 282(1):74-88.

2014

Neunzig J., Bernhardt R. (2014) Dehydroepiandrosterone sulfate (DHEAS) stimulates the first step in the biosynthesis of steroid hormones. J PLoS One. 9(2).

Brill E., Hannemann F., Zapp J., Brünig G., Jauch J., Bernhardt R. (2014) A new cytochrome P450 system from Bacillus megaterium DSM319 for the hydroxylation of 11-keto-beta-boswellic acid (KBA). Appl Microbiol Biotechnol 98(4):1701-1717.

2013

Khatri Y., Hannemann F., Girhard M., Kappl R., Meme A., Ringle M., Janocha S., Leize-Wagner E., Urlacher VB., Bernhardt R.(2013) Novel family members of CYP109 from Sorangium cellulosum So ce56 exhibit characteristic biochemical and biophysical properties. Biotechnol Appl Biochem. 60(1):18-29.

Janocha S., Bernhardt R.(2013) Design and characterization of an efficient CYP105A1-based whole-cell biocatalyst for the conversion of resin acid diterpenoids in permeabilized Escherichia coli. Appl Microbiol Biotechnol 97(17):7639-7649.

Elkalifa A.E., Bernhardt R. (2013)
Some physicochemical properties of flour from germinated sorghum grain. J. Food Sci. Technol. 50:186-190.

Ringle M., Khatri Y., Zapp J., Hannemann F., Bernhardt R. (2012) Application of a new versatile electron transfer system for cytochrome P450-based Escherichia coli whole-cell bioconversions. Appl. Microbiol. Biotechnol. 97(17): 7741-7754.

2012

Hobler A., Kagawa N., Hutter M.C., Hartmann M.F., Wudy S.A., Hannemann F., Bernhardt R. (2012) Human aldosterone synthase: recombinant expression in E. coli and purification enables a detailed biochemical analysis of the protein on the molecular level. J. Steroid Biochem. Mol. Biol. 132:57-65.

Schumacher S.D., Hannemann F., Teese M.G., Bernhardt R., Jose J. (2012) Autodisplay of functional CYP106A2 in Escherichia coli. J. Biotechnol. 161:104-12.2012

Ly T. T. B., Khatri Y., Zapp J., Hutter M. C., Bernhardt R. (2012) CYP264B1 from Sorangium cellulosum So ce56: a fascinating norisoprenoid and sesquiterpene hydroxylase. Eur. J. Appl. Microbiol. Biotechnol. 95: 123-133.

Ewen K. M., Ringle M., Bernhardt R. (2012) Adrenodoxin - A versatile ferredoxin. IUBMB Life 64: 506-512.

Nguyen K. T., Virus C., Günnewich N., Hannemann F., Bernhardt R. (2012) Changing the Regioselectivity of a P450 from C15 to C11 Hydroxylation of Progesterone. Chembiochem 13: 1161-1166.

Schmitz D., Zapp J., Bernhardt R. (2012) Hydroxylation of the triterpenoid dipterocarpol with CYP106A2 from Bacillus megaterium. FEBS Journal 279: 1663-1674.

Eiden-Plach A., Nguyen H. H., Schneider U., Hartmann M. F., Bernhardt R., Hannemann F., Wudy S. A. (2012) Alu Sx repeat-induced homozygous deletion of the StAR gene causes lipoid congenital adrenal hyperplasia. J. Steroid Biochem. Mol. Biol. 130: 1-6.

Wörner M., Melchior K., Monostory K., Pascussi J. M., Huber C. G., Bernhardt R. (2012) The Effects of Rosuvastatin and the CYP51A1 Inhibitor LEK-935 on the Proteome of Primary Human Hepatocytes. Drug Metab. Dispos. 40: 414-418.

Kleser M., Hannemann F., Hutter M., Zapp J., Bernhardt R. (2012) CYP105A1 mediated 3-hydroxylation of glimepiride and glibenclamide using a recombinant Bacillus megaterium whole-cell catalyst. J. Biotechnol. 157: 405-412.

Bleif S., Hannemann F., Zapp J., Hartmann D., Jauch J., Bernhardt R. (2012) A new Bacillus megaterium whole-cell catalyst for the hydroxylation of the pentacyclic triterpene 11-keto-beta-boswellic acid (KBA) based on a recombinant cytochrome P450 system. Eur. J. Appl. Microbiol. Biotechnol. 93: 1135-1146.

2011

Schiffler B., Zöllner A., Bernhardt R. (2011) Kinetic and optical biosensor study of adrenodoxin mutant AdxS112W displaying an enhanced interaction towards the cholesterol side chain cleavage enzyme (CYP11A1). Eur. Biophys. J. Biophy. 40: 1275-1282.

Ewen K. M., Hannemann F., Lametti S., Morleo A., Bernhardt R. (2011) Functional Characterization of Fdx1: Evidence for an Evolutionary Relationship between P450-Type and ISC-Type Ferredoxins. J. Mol. Biol. 413: 940-951.

Müller J. J., Hannemann F., Schiffler B., Ewen K. M., Kappl R., Heinemann U., Bernhardt R. (2011) Structural and thermodynamic characterization of the adrenodoxin-like domain of the electron-transfer protein Etp1 from Schizosaccharomyces pombe. J. Inorg. Biochem. 105: 957-965.

Khatri Y., Hannemann F., Perlova O., Müller R., (2011) and Bernhardt R. Investigation of cytochromes P450 in myxobacteria: Excavation of cytochromes P450 from the genome of Sorangium cellulosum So ce56. FEBS Letters 585: 1506-1513.

Hakki T., Hubel K., Waldmann H., Bernhardt R. (2011) The development of a whole-cell based medium throughput screening system for the discovery of human aldosterone synthase (CYP11B2) inhibitors: Old drugs disclose new applications for the therapy of congestive heart failure myocardial fibrosis and hypertension. J. Steroid Biochem. Mol. Biol. 125: 120-128.

Bleif S., Hannemann F., Lisurek M., von Kries J. P., Zapp J., Dietzen M., Antes I., Bernhardt R. (2011) Identification of CYP106A2 as a Regioselective Allylic Bacterial Diterpene Hydroxylase. Chembiochem 12: 576-582.

Tin M. K., Hakki T., Bernhardt R. (2011) Fission yeast Schizosaccharomyces pombe as a new system for the investigation of corticosterone methyloxidase deficiency-causing mutations. J. Steroid Biochem. Mol. Biol. 124: 31-37.

Garcia L. L., Rivas-Marin E., Floriano B., Bernhardt R., Ewen K. M., Reyes-Ramirez F., Santero E. (2011) ThnY Is a Ferredoxin Reductase-like Iron-Sulfur Flavoprotein That Has Evolved to Function as a Regulator of Tetralin Biodegradation Gene Expression. J. Biol. Chem. 286: 1709-1718.

Ewen K. M., Kleser M., Bernhardt R. (2011) Adrenodoxin: The archetype of vertebrate-type [2Fe-2S] cluster ferredoxins. Biochim. Biophys. Acta-Proteins and Proteomics 1814: 111-125.

Janocha S., Bichet A., Zöllner A., Bernhardt R. (2011) Substitution of lysine with glutamic acid at position 193 in bovine CYP11A1 significantly affects protein oligomerization and solubility but not enzymatic activity. Biochim. Biophys. Acta-Proteins and Proteomics 1814: 126-131.

Ivanov Y. D., Frantsuzov P. A., Zöllner A., Medvedeva N. V., Archakov A. I., Reinle W., Bernhardt R. (2011) Atomic Force Microscopy Study of Protein-Protein Interactions in the Cytochrome CYP11A1 (P450scc)-Containing Steroid Hydroxylase System. Nanoscale Res. Lett. 6: 54.

2010

Khatri Y., Hannemann F., Ewen K. M., Pistorius D., Perlova O., Kagawa N., Brachmann A. O., Müller R., Bernhardt R. (2010) The CYPome of Sorangium cellulosum So ce56 and Identification of CYP109D1 as a New Fatty Acid Hydroxylase. Chemistry & Biology 17: 1295-1305.

Petric S., Hakki T., Bernhardt R., Zigon D., Cresnar B. (2010) Discovery of a steroid 11 alpha-hydroxylase from Rhizopus oryzae and its biotechnological application. J. Biotechnol. 150: 428-437.

Khatri Y., Girhard M., Romankiewicz A., Ringle M., Hannemann F., Urlacher V. B., Hutter M. C., Bernhardt R. (2010) Regioselective hydroxylation of norisoprenoids by CYP109D1 from Sorangium cellulosum So ce56. Appl. Microbiol. Biotechnol. 88: 485-495.

Keizers P. H. J., Mersinli B., Reinle W., Donauer J., Hiruma Y., Hannemann F., Overhand M., Bernhardt R., Ubbink M. (2010) A Solution Model of the Complex Formed by Adrenodoxin and Adrenodoxin Reductase Determined by Paramagnetic NMR Spectroscopy. Biochemistry 49: 6846-6855.

Nguyen H. H., Hannemann F., Hartmann M. F., Malunowicz E. M., Wudy S. A., Bernhardt R. (2010) Five novel mutations in CYP11B2 gene detected in patients with aldosterone synthase deficiency type I: Functional characterization and structural analyses. Mol. Gen. Metab. 100: 357-364.

Elkhalifa A. E. O., Bernhardt R. (2010) Influence of grain germination on functional properties of sorghum flour. Food Chem. 121: 387-392.

Sheftel A. D., Stehling O., Pierik A. J., Elsasser H. P., Muhlenhoff U., Webert H., Hobler A., Hannemann F., Bernhardt R., Lill R. (2010) Humans possess two mitochondrial ferredoxins Fdx1 and Fdx2, with distinct roles in steroidogenesis heme Fe/S cluster biosynthesis. Proc. Natl. Acad. Sci. U.S.A. 107: 11775-11780.

Girhard M., Klaus T., Khatri Y., Bernhardt R., Urlacher V. B. (2010) Characterization of the versatile monooxygenase CYP109B1 from Bacillus subtilis. Appl. Microbiol. Biotechnol. 87: 595-607.

Zehentgruber D., Hannemann F., Bleif S., Bernhardt R., Lutz S. (2010) Towards Preparative Scale Steroid Hydroxylation with Cytochrome P450 Monooxygenase CYP106A2. Chembiochem 11: 713-721.

Berwanger A., Eyrisch S., Schuster I., Helms V., Bernhardt R. (2010) Polyamines: Naturally occurring small molecule modulators of electrostatic protein-protein interactions. J. Inorg. Biochem. 104: 118-125.

2009

Goni G., Zöllner A., Lisurek M., Velazquez-Campoy A., Pinto S., Gomez-Moreno C., Hannemann F., Bernhardt R., Medina M. (2009) Cyanobacterial electron carrier proteins as electron donors to CYP106A2 from Bacillus megaterium ATCC 13368. Biochim. Biophys. Acta -Proteins and Proteomics 1794: 1635-1642.

Ewen K. M., Hannemann F., Khatri Y., Perlova O., Kappl R., Krug D., Huttermann J., Müller R., Bernhardt R. (2009) Genome Mining in Sorangium cellulosum So ce56 IDENTIFICATION AND CHARACTERIZATION OF THE HOMOLOGOUS ELECTRON TRANSFER PROTEINS OF A MYXOBACTERIAL CYTOCHROME P450. J. Biol. Chem. 284: 28590-28598.

Lauer B. J., Worner M., Bernhardt R. (2009) Cloning and heterologous expression in Escherichia coli of the fission yeast vip1 gene showing differential expression after aldosterone treatment. C.R. Chim. 12: 1127-1139.

Keizers P., Mersinli B., Reinle W., Hiruma Y., Hannemann F., Bernhardt R., Overhand M., Ubbink M. (2009) The adrenodoxin redox chain protein complexes studied by paramagnetic NMR. FEBS J. 276: 144-144.

Hannemann F., Guyot A., Zöllner A., Müller J. J., Heinemann U., Bernhardt R. (2009) The dipole moment of the electron carrier adrenodoxin is not critical for redox partner interaction and electron transfer. J. Inorg. Biochem. 103: 997-1004.

Bernhardt R. (2009) Changing substrate specificity and activity of bacterial and mitochondrial P450 systems. FEBS Journal 276: 25-26.

Xu X. F., Keizers P. H. J., Reinle W., Hannemann F., Bernhardt R., Ubbink M. (2009)Intermolecular dynamics studied by paramagnetic tagging. J. Biomol. NMR 43: 247-254.

Worner M., Melchior K., Delmotte N., Hwang K. H., Monostory K., Huber C. G., Bernhardt R. (2009) Shotgun proteomic analysis of the microsomal fraction of eukaryotic cells using a two-dimensional reversed-phase x ion-pair reversed-phase HPLC setup. J. Sep. Sci. 32: 1165-1174.

Lovas K., McFarlane I., Nguyen H. H., Curran S., Schwabe J., Halsall D., Bernhardt R., Wallace A. M., Chatterjee V. K. K. J. (2009) Gene Mutation in an Asian Family with Aldosterone Synthase Deficiency. Clin. Endocrinol. Metab. 94: 914-919.

Monostory K., Pascussi J. M., Szabo P., Temesvari M., Kohalmy K., Acimovic J., Kocjan D., Kuzman D., Wilzewski B., Bernhardt R., Kobori L., Rozman D. (2009) Drug Interaction Potential of 2-((3,4-Dichlorophenethyl)(propyl)amino)-1-(pyridin-3-yl)ethanol (LK-935), the Novel Nonstatin-Type Cholesterol-Lowering Agent. Drug Metab. Dispos. 37: 375-385.

Hartmann M. F., Malunowicz E. M., Horn T., Zimmer K. P., Nguyen H. H., Hannemann F., Bernhardt R., Wudy S. A. (2009) Delineation of disorders of aldosterone metabolism from spot urine in early infancy by GC-MS steroid metabolomics. Horm. Res. 72: 75-76.

2008

Nguyen H.H., Hannemann F., Hartmann M.F., Wudy S.A., Bernhardt R. Aldosterone synthase deficiency caused by a homozygous L451F mutation in the CYP11B2 gene. Mol. Genet. Metab. 93: 458-67.

Zöllner A., Kagawa N., Waterman M.R., Nonaka Y., Takio K., Shiro Y., Hannemann F., Bernhardt R. (2008) Purification and functional characterization of human 11beta hydroxylase expressed in Escherichia coli. FEBS J. 275: 799-810.

Zöllner S., Hwang K.H., Wilzewski B., Carapito C., Leize-Wagner E., Van Dorsselaer A., Bernhardt R. (2008) Aldosterone: from biosynthesis to non-genomic action onto the proteome. Steroids 73: 966-72.

Ewen K.M., Schiffler B., Uhlmann-Schiffler H., Bernhardt R., Hannemann F. (2008) The endogenous adrenodoxin reductase-like flavoprotein arh1 supports heterologous cytochrome P450-dependent substrate conversions in Schizosaccharomyces pombe. FEMS Yeast Res. 8: 432-41.

Xu X., Reinle W., Hannemann F., Konarev P.V., Svergun D.I., Bernhardt R., Ubbink M. (2008) Dynamics in a pure encounter complex of two proteins studied by solution scattering and paramagnetic NMR spectroscopy. J. Am. Chem. Soc. 130: 6395-403.

Lisurek M., Simgen B., Antes I., Bernhardt R. (2008) Theoretical and experimental evaluation of a CYP106A2 low homology model and production of mutants with changed activity and selectivity of hydroxylation. Chembiochem 9: 1439-49.

Riedl S., Nguyen H.H., Clausmeyer S., Schulze E., Waldhauser F., Bernhardt R. (2008) A homozygous L299P mutation in the CYP11B1 gene leads to complete virilization in 46,XX individuals with 11-beta-hydroxylase deficiency. Horm. Res. 70: 145-149.

Ivanova L., Bernhardt R., Bernhardt I. (2008) Nongenomic effect of aldosterone on ion transport pathways of red blood cells. Cell Physiol Biochem. 22: 269-78.

Virus C., Bernhardt R. (2008) Molecular evolution of a steroid hydroxylating cytochrome P450 using a versatile steroid detection system for screening. Lipids 43: 1133-41.

2007

Behlke J., Ristau O., Müller E.C., Hannemann F., Bernhardt R. (2007) Self-association of adrenodoxin studied by using analyticalultracentrifugation.Biophys. Chem. 125: 159-65.

Hannemann F., Bichet a Ewen K.M., Bernhardt R. (2007) Cytochrome P450 systems-biological variations of electron transport chains.Biochim. Biophys. Acta 1770: 330-44.

Zöllner A., Pasquinelli M.A., Bernhardt R., Beratan D.N. (2007) Protein phosphorylation and intermolecular electron transfer: A joint experimental and computational study of a hormone biosynthesis pathway. J. Am. Chem. Soc. 129: 4206-4216.

Waterman M.R., Bernhardt R., Hildebrandt A.B., Prough R.A. (2007) Drug Metabolism Reviews: Preface. Drug Metabolism Reviews 39: 251.

Schuster I., Bernhardt R. (2007) Inhibition of cytochromes P450: Existing and new promising therapeutic targets. Drug Metabolism Reviews 39: 481-499.

Napoli E., Morin D., Bernhardt R., Buckpitt A., Cortopassi G. (2007) Hemin rescues adrenodoxin heme a and cytochrome oxidase activity in frataxin-deficient oligodendroglioma cells. Biochim. Biophys. Acta - Molecular Basis of Disease. 1772: 773-780.

Hannemann F., Bernhardt R., Jose J. (2007) Biocatalytic synthesis of 4-pregnen-20,21-diol-3-one a selectiveinhibitor of human 5alpha-reductase type II. Journal of Enzyme Inhibition and Medicinal Chemistry 22: 570-576.

Bichet A., Hannemann F., Rekowski M., Bernhardt R. (2007) A new application of the yeast two-hybrid system in protein engineering.Protein Eng. Des. Sel. 20: 117-123.

Freel E.M., Bernhardt R., Ingram M., Wallace A.M., Fraser R., Davies E., Connell J.M. (2007) Endogenous corticosteroid biosynthesis in subjects after bilateral adrenalectomy. Clin. Endocrinol. (Oxf) 66: 659-65.

2006

Kappl R., Ebelshäuser M., Hannemann F., Bernhardt R., Hüttermann J. (2006) Probing Electronic and Structural Properties of the Reduced [2Fe-2S] Cluster by Orientation-Selective 1H ENDOR Spectroscopy: Adrenodoxin versus Rieske Iron-Sulfur Protein. Appl. Magn. Reson. 30: 427-59.

Virus C., Lisurek M., Simgen B., Hannemann F., Bernhardt R. (2006) Function and engineering of the 15beta-hydroxylase CYP106A2. Biochem. Soc. Trans. 34: 1215-8.

Böhmer S., Carapito C., Wilzewski B., Leize E., Van Dorsselaer A., Bernhardt R. (2006) Analysis of aldosterone-induced differential receptor-independent proteinpatterns using 2D-electrophoresis and mass spectrometry. Biol. Chem. 387: 917-29.

Bauer S., Adrian N., Fischer E., Kleber S., Stenner F., Wadle A., Fadle N.,Zöllner A., Bernhardt R., Knuth A., Old L.J., Renner C. (2006) Structure-activity profiles of Ab-derived TNF fusion proteins. J. Immunol. 177: 2423-30.

Chung J.W., Park J.M., Bernhardt R., Pyun J.C. (2006) Immunosensor with a controlled orientation of antibodies by using NeutrAvidin-protein A complex at immunoaffinity layer. J. Biotechnol. 126: 325-33.

Hwang K.H., Carapito C., Böhmer S., Leize E., Van Dorsselaer A., Bernhardt R. (2006) Proteome analysis of Schizosaccharomyces pombe by two-dimensional gelelectrophoresis and mass spectrometry. Proteomics 6: 4115-29.

Chung J.W., Bernhardt R., Pyun J.C. (2006) Sequential analysis of multiple analytes using a surface plasmon resonance(SPR) biosensor. J. Immunol. Methods 311: 178-88.

Bernhardt R. (2006) Cytochromes P450 as versatile biocatalysts. J. Biotechnol. 124: 128-45.

Hannemann F., Virus C., Bernhardt R. (2006) Design of an Escherichia coli system for whole cell mediated steroid synthesisand molecular evolution of steroid hydroxylases. J. Biotechnol. 124: 172-81.

Hakki T., Bernhardt R. (2006) CYP17- and CYP11B-dependent steroid hydroxylases as drug development targets. Pharmacol. Ther. 111: 27-52.

Halavaty A., Müller J.J., Contzen J., Jung C., Hannemann F., Bernhardt R.,Galander M., Lendzian F., Heinemann U. (2006) Light-induced reduction of bovine adrenodoxin via the covalently boundruthenium(II) bipyridyl complex: intramolecular electron transfer and crystal structure. Biochemistry 45:709-18.

Elkhalifa A.E.O., Schiffler B., Bernhardt R. (2006) Some physico-chemical properties of fermented sorghum flour. Journal of Food Science and Technology 43: 26-27.

2005

Bureik M., Mion A., Kenyon C.J., Bernhardt R. (2005)Inhibition of aldosterone biosynthesis by staurosporine. Biol. Chem. 386: 663-9.

Heinz A., Hannemann F., Müller J.J., Heinemann U., Bernhardt R. (2005) The interaction domain of the redox protein adrenodoxin is mandatory for bindingof the electron acceptor CYP11A1, but is not required for binding of the electrondonor adrenodoxin reductase. Biochem. Biophys. Res. Commun. 338: 491-8.

Diesel B., Radermacher J., Bureik M., Bernhardt R., Seifert M., Reichrath J.,Fischer U., Meese E. (2005) Vitamin D(3) metabolism in human glioblastoma multiforme: functionality of CYP27B1splice variants metabolism of calcidiol effect of calcitriol. Clin. Cancer Res. 11: 5370-80.

Bureik M., Brück N., Hübel K., Bernhardt R. (2005) The human mineralocorticoid receptor only partially differentiates between differentligands after expression in fission yeast. FEMS Yeast Res. 5: 627-33.

Dragan C.A., Zearo S., Hannemann F., Bernhardt R., Bureik M. (2005) Efficient conversion of 11-deoxycortisol to cortisol (hydrocortisone)by recombinant fission yeast Schizosaccharomyces pombe. FEMS Yeast Res. 5: 621-5.

Ulmschneider S., Müller-Vieira U., Mitrenga M., Hartmann R.W., Oberwinkler-Marchais S., Klein C.D., Bureik M., Bernhardt R., Antes I., Lengauer T. (2005) Synthesis and evaluation of imidazolylmethylenetetrahydronaphthalenesand imidazolylmethyleneindanes: potent inhibitors of aldosterone synthase. J. Med. Chem. 48: 1796-805.

Bureik M., Zöllner A., Schuster N., Montenarh M., Bernhardt R. (2005) Phosphorylation of Bovine Adrenodoxin by Protein Kinase CK2 Affects the Interaction with Its Redox Partner Cytochrome P450(scc) (CYP11A1). Biochemistry 44: 3821-30.

2004

Kang M.J., Lisurek M., Bernhardt R., Hartmann R.W. (2004) Use of high-performance liquid chromatography/electrospray ionization collision-induced dissociation mass spectrometry for structural identification of monohydroxylated progesterones. Rapid Commun. Mass Spectrom. 18: 2795-800.

Elkalifa A.E., Schiffler B. Bernhardt R. (2004) Effect of fermentation on the starch digestibility resistant starch and some physicochemical properties of sorghum flour. Nahrung 48: 91-4.

Bernhardt R. (2004) Optimized chimeragenesis; creating diverse p450 functions. Chem. Biol. 11: 287-288.

Zöllner A., Nogues I., Heinz A., Medina M., Gomez-Moreno C., Bernhardt R. (2004) Analysis of the interaction of a hybrid system consisting of bovine adrenodoxin reductase and flavodoxin from the cyanobacterium Anabaena PCC 7119. Bioelectrochemistry 63: 61-65.

Bureik M., Hübel K., Dragan C.A., Scher J., Becker H., Lenz N., Bernhardt R. (2004) Development of test systems for the discovery of selective human aldosterone synthase (CYP11B2) and 11beta-hydroxylase (CYP11B1) inhibitors. Discovery of a new lead compound for the therapy of congestive heart failure myocardial fibrosis and hypertension. Mol. Cell. Endocrinol. 217: 249-254.

Bichet A., Bureik M., Lenz N., Bernhardt R. (2004) The "Bringer" strategy: a very fast and highly efficient method for construction of mutant libraries by error-prone polymerase chain reaction of ring-closed plasmids. Appl. Biochem. Biotechnol. 117: 115-122.

Lisurek M., Kang M.J., Hartmann R.W., Bernhardt R. (2004) Identification of monohydroxy progesterones produced by CYP106A2 using comparative HPLC and electrospray ionisation collision-induced dissociation mass spectrometry. Biochem. Biophys. Res. Commun. 319: 677-682.

Schiffler B., Zöllner A., Bernhardt R. (2004) Stripping down the mitochondrial cholesterol hydroxylase system - A kinetic study. J. Biol. Chem. 279: 34269-76.

Schiffler B., Bureik M., Reinle W., Müller E.C., Hannemann F., Bernhardt R. (2004) The adrenodoxin-like ferredoxin of Schizosaccharomyces pombe mitochondria. J. Inorg. Biochem. 98: 1229-1237.

Lisurek M., Bernhardt R. (2004) Modulation of aldosterone and cortisol synthesis on the molecular level. Mol. Cell. Endocrinol. 215: 149-159.

2003

Cereda A., Forlani F., Iametti S., Bernhardt R., Ferranti P., Picariello G., Pagani S., Bonomi F. (2003) Molecular recognition between Azotobacter vinelandii rhodanese and a sulfur acceptor protein. Biol. Chem. 384: 1473-1481.

Schiffler B., Bernhardt R. (2003) Bacterial (CYP101) and mitochondrial P450 systems-how comparable are they? Biochem. Biophys. Res. Commun. 312: 223-228, Review.

Kostic M., Bernhardt R., Pochapsky T.C. (2003) A conserved histidine in vertebrate-type ferredoxins is critical for redox-dependent dynamics. Biochemistry 42: 8171-8182.

Worrall J.A., Reinle W., Bernhardt R., Ubbink M. (2003) Transient protein interactions studied by NMR spectroscopy: the case of cytochrome C and adrenodoxin. Biochemistry 42: 7068-7076.

Strauch B., Zelinka T., Hampf M., Bernhardt R., Widimsky J. (2003) Jr. Prevalence of primary hyperaldosteronism in moderate to severe hypertension in the Central Europe region. J. Hum. Hypertens. 17: 349-352.

Faro M., Schiffler B., Heinz A., Nogues I., Medina M., Bernhardt R., Gomez-Moreno C. (2003) Insights into the design of a hybrid system between Anabaena ferredoxin-NADP+ reductase and bovine adrenodoxin. Eur. J. Biochem. 270: 726-735.

Bureik M., Zeeh A., Bernhardt R. (2003) Modulation of steroid hydroxylase activity in stably transfected V79MZh11B1 and V79MZh11B2 cells by PKC and PKD inhibitors. Endocr. Res. 28: 351-355.

Liakos P., Lenz D., Bernhardt R., Feige J.J., Defaye G. (2003) Transforming growth factor beta1 inhibits aldosterone and cortisol production in the human adrenocortical cell line NCI-H295R through inhibition of CYP11B1 and CYP11B2 expression. J. Endocrinol. 176: 69-82.

2002

Bechtel S., Belkina N., Bernhardt R. (2002) The effect of amino-acid substitutions l112p D147E and K152N in CYP11B2 on the catalytic activities of the enzyme. Eur. J. Biochem. 269: 1118-1127.

Lloyd-MacGilp S.A. , Torielli L., Bechtel S., Tripodi G., Gomez-Sanchez C., Zagato L., Bernhardt R., Kenyon C.J. (2002) Mutations in aldosterone synthase gene of Milan hypertensive rats: phenotypic consequences. Am. J. Physiol. Metab. 282: 608-617.

Bureik M., Schiffler B., Hiraoka Y., Vogel F. (2002), Bernhardt R. Functional expression of human mitochondrial CYP11B2 in fission yeast and identification of a new internal electron transfer protein etp1. Biochemistry 41: 2311-2321.

Bülow H.E., Bernhardt R. (2002) Analyses of the CYP11B gene family in the guinea pig suggest the existence of a primordial CYP11B gene with aldosterone synthase activity. Eur. J. Biochem. 269: 3838-3846.

Ehmer P.B., Bureik M., Bernhardt R., Müller U., Hartmann R.W. (2002) Development of a test system for inhibitors of human aldosterone synthase (CYP11B2): screening in fission yeast and evalutation of selectivity in V79 cells. Journal of Steroid Biochemistry & Molecular Biology 81: 173-179.

Jose J., Bernhardt R., Hannemann F. (2002) Cellular surface display of dimeric Adx and whole cell P450-mediated steroid synthesis on E. coli. Journal of Biotechnology 95: 257-268.

Beilke D., Weiss R., Löhr F., Pristovsek P. , Hannemann F., Bernhardt R., Rüterjans H. (2002) A new electron transport mechanism in mitochondrial steroid hydroxylase systems based on structural changes upon the reduction of adrenodoxin. Biochemistry 41: 7969-7978.

Hannemann F. Bera A.K., Fischer B., Lisurek M., Teuchner K., Bernhardt R. (2002) Unfolding and conformational studies on bovine adrenodoxin probed by engineered intrinsic tryptophan fluorescence. Biochemistry 41: 11008-11016.

Zöllner A., Hannemann F., Lisurek M., Bernhardt R. (2002) Deletions in the loop surrounding the iron-sulfur cluster of adrenodoxin severely affect the interactions with its native redox partners adrenodoxin reductase and cytochrome P450scc (CYP11A1). J. Inorg. Biochem. 91: 644-654.

Bureik M., Lisurek M., Bernhardt R. (2002) The human steroid hydroxylases CYP11B1 and CYP11B2.Biol. Chem., 383: 1537-1551.

2001

Hannemann F., Rottmann M., Schiffler B., Bernhardt R. (2001) The loop region covering the iron-sulfur cluster in bovine adrenodoxin comprises a new interaction site for redox partners. J. Biol. Chem. 276: 1369-1375.

Iametti S., Bera A. K., Vecchio G., Grinberg A., Bernhardt R., Bonomi F. (2001) GroEL-assisted refolding of adrenodoxin during chemical cluster insertion. Eur. J. Biochem. 268: 2421-2429.

Jose J., Bernhardt R., Hannemann F. (2001) Functional display of active adrenodoxin a [ 2Fe-2S] ferredoxin on the surface of E. coli. ChemBioChem 2: 695-701.

Schiffler B. , Kiefer M., Wilken A., Hannemann F., Adolph H.W., Bernhardt R. (2001) The interaction of bovine adrenodoxin with CYP11A1 (cytochrome P450scc) and CYP11B1 (cytochrome P45011ß): acceleration of reduction and substrate conversion by site-directed mutagenesis of adrenodoxin. J. Biol. Chem. 276: 36225-36232.

Grinberg A.V., Bernhardt R. (2001) Contribution of a Salt Bridge to the Thermostability of Adrenodoxin Determined by Site-Directed Mutagenesis. Archiv of Biochemistry and Biophysics 396: 25-34.

Hampf M., Ngoc Dao N.T., Hoan N.T. Bernhardt R.(2001) Unequal crossing-over between aldosterone synthase and 11b-hydroxylase genes causes congenital adrenal hyperplasia. J. Clin. Endocrinol. Metab. 86: 4445-4452.

Fisher A., Friel E.C., Bernhardt R., Gomez-Sanchez C., Mc Connell J., Fraser R., Davies E. (2001) Effects of 18-hydroxylated steroids on corticosteroid production by human aldosterone synthase and 11b-hydroxylase. J. Clin. Endocrinol. Metab. 86: 4326-4329.

Belkina N.V., Lisurek M., Ivanov A.S., Bernhardt R. (2001) Modelling of 3D-structures of cytochromes P450 11B1 AND 11B2. J. Inorg. Biochem. 87: 197-207.

2000

Cao P.R., Bülow H., Dumas B., Bernhardt R. (2000) Construction and characterization of a catalytic fusion protein system: P-450(11beta)-adrenodoxin reductase-adrenodoxin. Biochim. Biophys. Acta 1476: 253-264.

Simgen B., Contzen J., Schwarzer R., Bernhardt R., Jung C. (2000) Substrate binding to 15ß-Hydroxylase (CYP106A2) probed by FT Infrared spectroscopic studies on the iron ligand CO stretch vibration. Biochem. Biophys. Res. Commun. 269: 737-742.

Bernhardt R. (2000) The role of adrenodoxin in adrenal steroidogenesis. Curr. Opinion Endocrinol. & Diabetes 7: 109-115.

Grinberg A.V., Hannemann F., Schiffler B., Müller J.J., Heinemann U., Bernhardt R. (2000) Adrendoxin: structure stability and electron transfer properties. Proteins 40: 590-612.

Weiss R., Brachais L., Lohr F., Hartleib J., Bernhardt R., Rüterjans H. (2000) Assignment of 1h 13C and 15N signals of bovine adrenodoxin. J. Biomol. NMR 17: 355-356.

Bernhardt R., Cao P.R., Grinberg A., Hannemann F., Weißgerber P., Müller J.J., Heinemann U. (2000) Protein recognition in mitochondrial steroid hydroxylation and electron transport. in: Molecular Steroidogenesis (Okamoto M., Ishimura Y., Nawata H., Eds.) Universal Academy Press Inc. Tokyo Japan pp. 53-56.

Bechtel S., Hampf M., Bernhardt R. (2000) Influence of residues 112, 147 and 152 on the function of CYP11B2. in: Molecular Steroidogenesis (Okamoto M., Ishimura Y., Nawata H., Eds.) Universal Academy Press Inc. Tokyo Japan pp. 93-95.

Bureik M., Schiffler B., Rüdiger V., Vogel F., Bernhardt R. (2000) Fission yeast as a model system to study mitochondrial cytochrome P450 systems. in: Molecular Steroidogenesis (Okamoto M., Ishimura Y., Nawata H., Eds.) Universal Academy Press Inc. Tokyo Japan pp. 97-98.

Hannemann F., Bernhardt R. (2000) Identification of a new key position in the Adx molecule for interaction with its redox partners. in: Molecular Steroidogenesis (Okamoto M., Ishimura Y., Nawata H., Eds.) Universal Academy Press Inc. Tokyo Japan pp. 115-116.

Hampf M., Schulze E., Bernhardt R. (2000) Donor splice site mutation GTà GC in intron 8 of CYP11B1 causes steroid 11b -hydroxylase deficiency. in: Molecular Steroidogenesis (Okamoto M., Ishimura Y., Nawata H., Eds.) Universal Academy Press Inc. Tokyo Japan pp. 363.

Hampf M., Ngoc Dao N.T., Hoan N.T., Bernhardt R. (2000) New Molecular Genetic Defects causing 11ß-Hydroxylase Deficiency (CAH). Endocrine Research 26: 791-782.

1999

Lehnerer M., Schulze J., Bernhardt R., Hlavica P. (1999) Some properties of mitochondrial adrenodoxin associated with its nonconventional electron donor function toward rabbit liver microsomal cytochrome P450 2B4. Biochem. Biophys. Res. Commun. 254: 83-87.

Bera A.K., Grinberg A., Bernhardt R. (1999) A step toward understandíng the folding mechanism of bovine adrenodoxin. Arch. Biochem. Biophys. 361: 315-322.

Cao P., Bernhardt R. (1999) Interaction of CYP11B1 (cytochrome P45011ß) with CYP11A1 (cytochrome P450scc) in COS-1 cells. Eur. J. Biochem. 262: 720-726.

Bera A.K., Bernhardt R. (1999) GroEL assisted and unassisted refolding of mature and precursor adrenodoxin: the role of the precursor sequence. Arch. Biochem. Biophys. 367: 89-94.

Cao P., Bernhardt R. (1999) Modulation of aldosterone biosynthesis by adrenodoxin mutants with different electron transport efficiencies. Eur. J. Biochem. 265: 152-159.

Seemann T., Widimsky J., Hampf m Bernhardt R. (1999) Abolished nocturnal blood pressure fall in a boy with glucocorticoid-remediable aldosteronism. J. Human Hypertension 13: 823-828.

Müller J.J., Müller A., Rottmann M., Bernhardt R., Heinemann U. (1999) Vertebrate-type and plant-type ferredoxins: crystal structure comparison and electron transfer pathway modelling. J. Mol. Biol. 294: 501-513.

1998

Böttner B., Cao P., Bernhardt R. (1998) Changing substrate specificity and product pattern in adrenal cytochrome P450-dependent steroid hydroxylases. in: Oxygen Homeostasis and Its Dynamics Springer-Verlag Tokyo Berlin Heidelberg New York pp. 221-230.

Denner K., Bernhardt R. (1998) Inhibition studies of steroid conversions mediated by human CYP11B1and CYP11B2 expressed in cell cultures. in: Oxygen Homeostasis and Its Dynamics Springer-Verlag Tokyo Berlin Heidelberg New York pp. 231-236.

Peters J., Hampf M., Peters B., Bernhardt R. (1998) Molekularbiologie Klinik und Therapie steroidbedingter Hypertonien. in: Handbuch der Molekularen Medizin (D. Ganten und K. Ruckpaul Eds.) Springer-Verlag Berlin Heidelberg New York pp. 413-452.

Iametti S., Uhlmann H., Ragg E., Sala N., Beckert V., Grinberg A., Bernhardt R., Bonomi F. (1998) Cluster-iron substitution is related to structural and functional features of adrenodoxinmutants and to their redox states. Eur. J. Biochem. 251: 673-681.

Müller A., Müller J.J., Muller Y.A., Uhlmann H., Bernhardt R., Heinemann U. (1998) New aspects of electron transfer revealed by the crystal structure at of a truncated bovine adrenodoxin Adx(4-108). Structure 6: 269-280.

Böttner B., Denner K., Bernhardt R. (1998) Conferring aldosterone synthesis to human CYP11B1 by replacing key amino acid residues with CYP11B2-specific ones. Eur. J. Biochem. 252: 458-466.

Grinberg A., Bernhardt R. (1998) Structural and functional consequences of substitutions at the Pro108-Arg14 hydrogenbond in bovine adrenodoxin. Biochem. Biophys. Res. Commun. 249: 933-937.

Grinberg A., Bernhardt R. (1998) Effect of the replacing a conserved proline residue on the function and stability of bovine adrenodoxin. Protein Engineering 11: 1057-1064.

Goder V., Beckert V., Pfeil W., Bernhardt R. (1998) Impact of the presequence of a mitochondrium targeted precursor preadrenodoxinon folding catalytic activity and stability of the protein. Arch. Biochem. Biophys. 359: 31-41.

Bernhardt R., Müller A., Uhlmann H., Grinberg A., Müller J.J., Heinemann U. (1998) Structure of adrenodoxin and function in mitochondrial steroid hydroxylation. Endocrine Research 24: 531-539.

Hampf M., Widimsky J., Bernhardt R. (1998) Aldosterone synthase gene in patients suffering from hyperaldosteronism.Endocrine Research 24: 877-880.

Nonaka Y., Fujii T., Bernhardt R., Okamoto M. (1998) Amino acid residues in I- and K-helices of rat CYP11B1 and CYP11B2 are important in expression of 18-hydroxylation activity. Endocrine Research 24: 615-618.

Bernhardt R. (1998) Electron transfer in mitochondrial steroid hydroxylating cytochrome P450 systems: role of adrenodoxin. in: Biophysics of Electron Transfer and Molecular Bioelectronics (Nicolini C., Ed.) Plenum Press New York pp. 51-66.

Jin W., Wollenberger U., Bernhardt R., Stöcklein W.F.M., Scheller F.W. (1998) Direct electron transfer of adrenodoxin - a [2Fe-2S] protein - and its mutants atmodified gold electrode.Bioelectrochem. Bioenerg. 47: 75-79.

1997

Haseloff R.F., Zöllner S., Kirilyuk I.A., Grigorev I.A., Reszka R., Bernhardt R., Mertsch K., Roloff B., Blasig I.E. (1997)Superoxide-mediated reduction of the nitroxide group can prevent detection of nitric oxide by nitroyl nitroxides. Free Radical Res. 26: 7-17.

Beckert V., Bernhardt R. (1997) Specific aspects of electron transfer from adrenodoxin to cytochromes P450scc and P45011ß. J. Biol. Chem. 272: 4883-4888.

Uhlmann H., Iametti S., Vecchio G., Bonomi F., Bernhardt R. (1997) Proline 108 is important for folding and stabilization of adrenal ferredoxin but does not influence the functional properties of the protein. Eur. J. Biochem. 248: 897-902.

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