Download PDF
Horm Metab Res 2008; 40(11): 752-759
DOI: 10.1055/s-0028-1082039
Original Basic

© Georg Thieme Verlag KG Stuttgart · New York

Rosiglitazone Reduces Angiotensin II and Advanced Glycation End Product-dependent Sustained Nuclear Factor-κB Activation in Cultured Human Proximal Tubular Epithelial Cells

M. Morcos 1 , A. Schlotterer 1 , A. A. R. Sayed 1 , G. Kukudov 1 , D. Oikomonou 1 , Y. Ibrahim 1 , F. Pfisterer 1 , J. Schneider 1 , 2 , F. Bozorgmehr 1 , G. Rudofsky Jr. 1 , V. Schwenger 1 , R. Kientsch-Engels 3 , A. Hamann 1 , 6 , M. Zeier 1 , K. Dugi 4 , B. Yard 5 , P. M. Humpert 1 , F. van der Woude 5 , P. P. Nawroth 1 , A. Bierhaus 1
  • 1Department of Internal Medicine 1 (Endocrinology, Metabolism and Clinical Chemistry) & Renal Department, University of Heidelberg, Germany
  • 2Teaching Hospital Merzig/Saar, University of Saarland, Germany
  • 3Roche Diagnostics, Penzberg, Germany
  • 4Boehringer Ingelheim Corporation, Ingelheim, Germany
  • 5V Medical Clinic, University of Mannheim, Germany
  • 6Diabetes Clinic Bad Nauheim, Germany
Further Information

Publication History

received 08.01.2008

accepted 07.04.2008

Publication Date:
18 August 2008 (online)

Also available at
    Permissions and Reprints

Abstract

Tubular damage is a major feature in the development of diabetic nephropathy. This study investigates the effects of the thiazolidindione rosiglitazone on angiotensin II and advanced glycation end product-induced tubular activation in human proximal tubular epithelial cells in vitro. Angiotensin II and advanced glycation end products, both induced a dose-dependent sustained activation of the redox-sensitive transcription factor, Nuclear Factor kappa B (NF-κB). Nuclear translocation of NF-κB was evident already after one hour and persistent for more than four days. Co-incubation of proximal tubular epithelial cells with rosiglitazone significantly reduced angiotensin II and advanced glycation end product-mediated generation of reactive oxygen species, angiotensin II-dependent advanced glycation end product formation, NF-κB activation, and NF-κB-dependent pro inflammatory gene expression. Most importantly, rosiglitazone effects on NFκB activation were maximal at later time points, indicating that rosiglitazone treatment confers long lasting renoprotective effects.

Key words

cellular activation - late diabetic complication - diabetic nephropathy - thiazolidindiones - reactive oxygen species

References

  • 1 Becker BN, Kondo S, Cheng HF, Harris RC. Effect of glucose, pyruvate, and insulin on type 1 angiotensin II receptor expression in SV40-immortalized rabbit proximal tubule epithelial cells.  Kidney Int. 1997;  52 87-92
    CrossrefPubMedGoogle Scholar
  • 2 Gilbert RE, Cooper ME. The tubulointerstitium in progressive diabetic kidney disease: more than an aftermath of glomerular injury?.  Kidney Int. 1999;  56 1627-1637
    CrossrefPubMedGoogle Scholar
  • 3 Gomez-Garre D, Largo R, Tejera N, Fortes J, Manzarbeitia F, Egido J. Activation of NF-kappaB in tubular epithelial cells of rats with intense proteinuria: role of angiotensin II and endothelin-1.  Hypertension. 2001;  37 1171-1178
    PubMedGoogle Scholar
  • 4 Phillips AO. The role of renal proximal tubular cells in diabetic nephropathy.  Curr Diab Rep. 2003;  3 491-496
    CrossrefPubMedGoogle Scholar
  • 5 Saito A, Takeda T, Sato K, Hama H, Tanuma A, Kaseda R, Suzuki Y, Gejyo F. Significance of proximal tubular metabolism of advanced glycation end products in kidney diseases.  Ann N Y Acad Sci. 2005;  1043 637-643
    CrossrefPubMedGoogle Scholar
  • 6 Wang SN, LaPage J, Hirschberg R. Pathophysiologic glomerulotubular growth factor link.  Miner Electrolyte Metab. 1999;  25 234-241
    CrossrefPubMedGoogle Scholar
  • 7 Yaqoob M, MacClelland P, Patrick AW, Stevenson A, Mason H, Bell GM. Tubulopathy with macroalbuminuria due to diabetic nephropathy and primary glomerulonephritis.  Kidney Int. 1994;  47 101-104
    PubMedGoogle Scholar
  • 8 Yaqoob M, MacClelland P, Patrick AW, Stevenson A, Mason H, White MC, Bell GM. Evidence of oxidant injury and tubular damage in early diabetic nephropathy.  QJM. 1994;  87 601-607
    PubMedGoogle Scholar
  • 9 Ha H, Lee HB. Reactive oxygen species amplify glucose signalling in renal cells cultured under high glucose and in diabetic kidney.  Nephrology (Carlton). 2005;  10 ((Suppl)) S7-S10
    CrossrefPubMedGoogle Scholar
  • 10 Morcos M, Sayed AA, Bierhaus A, Yard B, Waldherr R, Merz W, Kloeting I, Schleicher E, Mentz S, Abd el Baki RF, Tritschler H, Kasper M, Schwenger V, Hamann A, Dugi KA, Schmidt AM, Stern D, Ziegler R, Haering HU, Andrassy M, Woude F van der, Nawroth PP. Activation of tubular epithelial cells in diabetic nephropathy.  Diabetes. 2002;  51 3532-3544
    CrossrefPubMedGoogle Scholar
  • 11 Tang SC, Leung JC, Chan LY, Tsang AW, Lai KN. Activation of tubular epithelial cells in diabetic nephropathy and the role of the peroxisome proliferator-activated receptor-gamma agonist.  J Am Soc Nephrol. 2006;  17 1633-1643
    CrossrefPubMedGoogle Scholar
  • 12 Ziyadeh FN. Significance of tubulointerstitial changes in diabetic renal disease.  Kidney Int Suppl. 1996;  54 10-13
    PubMedGoogle Scholar
  • 13 Başar Y, Salmayenli N, Aksoy M, Seçkin S, Aydin M, Ozkök E. ACE gene polymorphism in peripheral vascular disease.  Horm Metab Res. 2007;  3 534-537
    Article in Thieme ConnectPubMedGoogle Scholar
  • 14 Burns KD. Angiotensin II and its receptors in the diabetic kidney.  Am J Kidney Dis. 2000;  36 449-467
    CrossrefPubMedGoogle Scholar
  • 15 Diep QN, El Mabrouk M, Cohn JS, Endemann D, Amiri F, Virdis A, Neves MF, Schiffrin EL. Structure, endothelial function, cell growth, and inflammation in blood vessels of angiotensin II-infused rats: role of peroxisome proliferator-activated receptor-gamma.  Circulation. 2002;  105 2296-2302
    CrossrefPubMedGoogle Scholar
  • 16 Forbes JM, Cooper ME, Thallas V, Burns WC, Thomas MC, Brammar GC, Lee F, Grant SL, Burrell LA, Jerums G, Osicka TM. Reduction of the accumulation of advanced glycation end products by ACE inhibition in experimental diabetic nephropathy.  Diabetes. 2002;  51 3274-3282
    PubMedGoogle Scholar
  • 17 Forbes JM, Fukami K, Cooper ME. Diabetic nephropathy: where hemodynamics meets metabolism.  Exp Clin Endocrinol Diabetes. 2007;  115 69-84
    Article in Thieme ConnectPubMedGoogle Scholar
  • 18 Ruiz-Ortega M, Lorenzo O, Ruperez M, Blanco J, Egido J. Systemic infusion of angiotensin II into normal rats activates nuclear factor-kappaB and AP-1 in the kidney: role of AT(1) and AT(2) receptors.  Am J Pathol. 2001;  158 1743-1756
    PubMedGoogle Scholar
  • 19 Thomas MC, Tikellis C, Burns WM, Bialkowski K, Cao Z, Coughlan MT, Jandeleit-Dahm K, Cooper ME, Forbes JM. Interactions between renin angiotensin system and advanced glycation in the kidney.  J Am Soc Nephrol. 2005;  16 2976-2984
    CrossrefPubMedGoogle Scholar
  • 20 Wolf G. New insights into the pathophysiology of diabetic nephropathy: from haemodynamics to molecular pathology.  Eur J Clin Invest. 2004;  34 785-796
    CrossrefPubMedGoogle Scholar
  • 21 Nishikawa T, Edelstein D, Du XL, Yamagishi S, Matsumura T, Kaneda Y, Yorek MA, Beebe D, Oates PJ, Hammes HP, Giardino I, Brownlee M. Normalizing mitochondrial superoxide production blocks three pathways of hyperglycaemic damage.  Nature. 2000;  404 787-789
    CrossrefPubMedGoogle Scholar
  • 22 Gugliucci A, Bendayan M. Renal fate of circulating advanced glycated end products (AGE). Evidence for reabsorption and catabolism of AGE-peptide by renal proximal tubular cells.  Diabetologia. 1996;  39 149-160
    CrossrefPubMedGoogle Scholar
  • 23 Bierhaus A, Schiekofer S, Schwaninger M, Andrassy M, Humpert PM, Chen J, Hong M, Luther T, Henle T, Klöting I, Morcos M, Hofmann M, Tritschler H, Weigle B, Kasper M, Smith M, Perry G, Schmidt AM, Stern DM, Häring HU, Schleicher E, Nawroth PP. Diabetes-associated sustained activation of the transcription factor nuclear factor-kappaB.  Diabetes. 2001;  50 2792-2808
    CrossrefPubMedGoogle Scholar
  • 24 Bierhaus A, Humpert PM, Stern DM, Arnold B, Nawroth PP. Advanced glycation end product receptor-mediated cellular dysfunction.  Ann N Y Acad Sci. 2005;  1043 676-680
    CrossrefPubMedGoogle Scholar
  • 25 Kallwellis K, Grempler R, Günther S, Päth G, Walther R. Tumor necrosis factor alpha induces the expression of the nuclear protein p8 via a Novel NFkappaB binding site within the promoter.  Horm Metab Res. 2006;  38 570-574
    Article in Thieme ConnectPubMedGoogle Scholar
  • 26 Ruiz-Ortega M, Bustos C, Hernandez-Presa MA, Lorenzo O, Plaza JJ, Egido J. Angiotensin II participates in mononuclear cell recruitment in experimental immune complex nephritis through nuclear factor-kappa B activation and monocyte chemoattractant protein-1 synthesis.  J Immunol. 1998;  16 430-439
    PubMedGoogle Scholar
  • 27 Goldstein BJ. Rosiglitazone.  Int J Clin Pract. 2000;  54 333-337
    PubMedGoogle Scholar
  • 28 Efrati S, Berman S, Chachashvili A, Cohen N, Averbukh Z, Weissgarten J. Rosiglitazone treatment attenuates expression of inflammatory hallmarks in the remaining kidney following contralateral nephrectomy.  Am J Nephrol. 2007;  24 ((28)) 238-245
    PubMedGoogle Scholar
  • 29 Eldershaw TP, Rattigan S, Cawthorne MA, Buckingham RE, Colquhoun EQ, Clark MG. Treatment with the thiazolidinedione (BRL 49653) decreases insulin resistance in obese Zucker hindlimb.  Horm Metab Res. 1995;  2 169-172
    Article in Thieme ConnectPubMedGoogle Scholar
  • 30 Giorgini AE, Beales PE, Mire-Sluis A, Scott D, Liddi R, Pozzilli P. Troglitazone exhibits immunomodulatory activity on the cytokine production of activated human lymphocytes.  Horm Metab Res. 1999;  31 1-4
    Article in Thieme ConnectPubMedGoogle Scholar
  • 31 Panigrahy D, Singer S, Shen LQ, Butterfield CE, Freedman DA, Chen EJ, Moses MA, Kilroy S, Duensing S, Fletcher C, Fletcher JA, Hlatky L, Hahnfeldt P, Folkman J, Kaipainen A. PPARgamma ligands inhibit primary tumor growth and metastasis by inhibiting angiogenesis.  J Clin Invest. 2002;  110 923-932
    CrossrefPubMedGoogle Scholar
  • 32 Xin X, Yang S, Kowalski J, Gerritsen ME. Peroxisome proliferator-activated receptor gamma ligands are potent inhibitors of angiogenesis in vitro and in vivo.  J Biol Chem. 1999;  274 9116-9121
    CrossrefPubMedGoogle Scholar
  • 33 Lee S, Kim W, Kang KP, Moon SO, Sung MJ, Kim DH, Kim HJ, Park SK. Agonist of peroxisome proliferator-activated receptor-gamma, rosiglitazone, reduces renal injury and dysfunction in a murine sepsis model.  Nephrol Dial Transplant. 2005;  20 1057-1065
    CrossrefPubMedGoogle Scholar
  • 34 Sommer M, Wolf G. Rosiglitazone increases PPARgamma in renal tubular epithelial cells and protects against damage by hydrogen peroxide.  Am J Nephrol. 2007;  27 425-434
    CrossrefPubMedGoogle Scholar
  • 35 Göke B. Are current oral treatment options sufficient?.  Exp Clin Endocrinol Diabetes. 2000;  108 243-249
    Article in Thieme ConnectPubMedGoogle Scholar
  • 36 Pfützner A, Hohberg C, Lübben G, Pahler S, Pfützner AH, Kann P, Forst T. Pioneer study: PPARgamma activation results in overall improvement of clinical and metabolic markers associated with insulin resistance independent of long-term glucose control.  Horm Metab Res. 2005;  37 510-515
    Article in Thieme ConnectPubMedGoogle Scholar
  • 37 Dandona P, Aljada A. Endothelial dysfunction in patients with type 2 diabetes and the effects of thiazolidinedione antidiabetic agents.  J Diabet Complications. 2004;  18 91-102
    PubMedGoogle Scholar
  • 38 Sarafidis PA, Lasaridis AN, Nilsson PM, Hitoglou-Makedou AD, Pagkalos EM, Yovos JG, Pliakos CI, Tourkantonis AA. The effect of rosiglitazone on urine albumin excretion in patients with type 2 diabetes mellitus and hypertension.  Am J Hypertens. 2005;  18 227-234
    CrossrefPubMedGoogle Scholar
  • 39 Pistrosch F, Herbrig K, Kindel B, Passauer J, Fischer S, Gross P. Rosiglitazone improves glomerular hyperfiltration, renal endothelial dysfunction, and microalbuminuria of incipient diabetic nephropathy in patients.  Diabetes. 2005;  54 2206-2211
    CrossrefPubMedGoogle Scholar
  • 40 Raptis AE, Viberti G. Pathogenesis of diabetic nephropathy.  Exp Clin Endocrinol Diabetes. 2001;  109 424-437
    PubMedGoogle Scholar
  • 41 Iglesias P, Díez JJ. Peroxisome proliferator-activated receptor gamma agonists in renal disease.  Eur J Endocrinol. 2006;  154 613-621
    CrossrefPubMedGoogle Scholar
  • 42 Gross P, Pistrosch F. PPAR agonists in diabetic nephropathy.  Nephrol Dial Transplant. 2007;  22 2095
    PubMedGoogle Scholar
  • 43 Detrisac CJ, Sens MA, Garvin AJ, Spicer SS, Sens DA. Tissue culture of human kidney epithelial cells of proximal tubule origin.  Kidney Int. 1984;  25 383-390
    CrossrefPubMedGoogle Scholar
  • 44 Helbert MJ, Dauwe SE, Biest I Van der, Nouwen EJ, Broe ME De. Immunodissection of the human proximal nephron: flow sorting of S1S2S3, S1S2 and S3 proximal tubular cells.  Kidney Int. 1997;  52 414-428
    CrossrefPubMedGoogle Scholar
  • 45 Phillips AO, Steadman R, Topley N, Williams JD. Elevated D-glucose concentrations modulate TGF-beta 1 synthesis by human cultured renal proximal tubular cells. The permissive role of platelet-derived growth factor.  Am J Pathol. 1995;  147 362-374
    PubMedGoogle Scholar
  • 46 Sens DA, Detrisac CJ, Sens MA, Rossi MR, Wenger SL, Todd JH. Tissue culture of human renal epithelial cells using a defined serum-free growth formulation.  Exp Nephrol. 1999;  344-352
    CrossrefPubMedGoogle Scholar
  • 47 Bierhaus A, Illmer T, Kasper M, Luther T, Quehenberger P, Tritschler H, Wahl P, Ziegler R, Muller M, Nawroth PP. Advanced glycation end product (AGE)-mediated induction of tissue factor in cultured endothelial cells is dependent on RAGE.  Circulation. 1997;  96 2262-2271
    PubMedGoogle Scholar
  • 48 Bierhaus A, Ritz E, Nawroth PP. Expression of receptors for advanced glycation end-products in occlusive vascular and renal disease.  Nephrol Dial Transplant. 1996;  5 87-90
    PubMedGoogle Scholar
  • 49 Wagner Z, Wittmann I, Mazak I, Schinzel R, Heidland A, Kientsch-Engel R, Nagy J. N (epsilon)-(carboxymethyl) lysine levels in patients with type 2 diabetes: role of renal function.  Am J Kidney Dis. 2000;  38 785-791
    PubMedGoogle Scholar
  • 50 Piconi L, Quagliaro L, Ceriello A. Oxidative stress in diabetes.  Clin Chem Lab Med. 2003;  41 1144-1149
    CrossrefPubMedGoogle Scholar
  • 51 Bierhaus A, Humpert PM, Morcos M, Wendt T, Chavakis T, Arnold B, Stern DM, Nawroth PP. Understanding RAGE, the receptor for advanced glycation end products.  J Mol Med. 2005;  83 876-886
    CrossrefPubMedGoogle Scholar
  • 52 Bierhaus A, Stern DM, Nawroth PP. RAGE in inflammation: a new therapeutic target?.  Curr Opin Investig Drugs. 2006;  7 985-991
    PubMedGoogle Scholar
  • 53 Fukami K, Ueda S, Yamagishi S, Kato S, Inagaki Y, Takeuchi M, Motomiya Y, Bucala R, Iida S, Tamaki K, Imaizumi T, Cooper ME, Okuda S. AGEs activate mesangial TGF-beta-Smad signaling via an angiotensin II type I receptor interaction.  Kidney Int. 2004;  66 2137-2147
    CrossrefPubMedGoogle Scholar
  • 54 Forbes JM, Thomas MC, Thorpe SR, Alderson NL, Cooper ME. The effects of valsartan on the accumulation of circulating and renal advanced glycation end products in experimental diabetes.  Kidney Int Suppl. 2004;  92 S105-S107
    PubMedGoogle Scholar
  • 55 Forbes JM, Thorpe SR, Thallas-Bonke V, Pete J, Thomas MC, Deemer ER, Bassal S, El-Osta A, Long DM, Panagiotopoulos S, Jerums G, Osicka TM, Cooper ME. Modulation of soluble receptor for advanced glycation end products by angiotensin-converting enzyme-1 inhibition in diabetic nephropathy.  J Am Soc Nephrol. 2005;  16 2363-2372
    CrossrefPubMedGoogle Scholar
  • 56 Igarashi M, Takeda Y, Ishibashi N, Takahashi K, Mori S, Tominaga M, Saito Y. Pioglitazone reduces smooth muscle cell density of rat carotid arterial intima induced by balloon catheterization.  Horm Metab Res. 1997;  29 444-449
    Article in Thieme ConnectPubMedGoogle Scholar
  • 57 Rudofsky  Jr  G, Reismann P, Grafe IA, Konrade I, Djuric Z, Tafel J, Buchbinder S, Zorn M, Humpert PM, Hamann A, Morcos M, Nawroth PP, Bierhaus A. Improved vascular function upon pioglitazone treatment in type 2 diabetes is not associated with changes in mononuclear NF-kappaB binding activity.  Horm Metab Res. 2007;  39 665-671
    Article in Thieme ConnectPubMedGoogle Scholar
  • 58 Ihara Y, Egashira K, Nakano K, Ohtani K, Kubo M, Koga J, Iwai M, Horiuchi M, Gang Z, Yamagishi S, Sunagawa K. Upregulation of the ligand-RAGE pathway via the angiotensin II type I receptor is essential in the pathogenesis of diabetic atherosclerosis.  J Mol Cell Cardiol. 2007;  43 455-464
    CrossrefPubMedGoogle Scholar
  • 59 Marx N, Walcher D, Ivanova N, Rautzenberg K, Jung A, Friedl R, Hombach V, Caterina R de, Basta G, Wautier MP, Wautiers JL. Thiazolidinediones reduce endothelial expression of receptors for advanced glycation end products.  Diabetes. 2004;  53 2662-2668
    CrossrefPubMedGoogle Scholar
  • 60 Bierhaus A, Chevion S, Chevion M, Quehenberger P, Hofmann M, Illmer T, Luther T, Berentshtein E, Tritschler H, Müller M, Ziegler R, Nawroth PP. Advanced glycation endproducts (AGEs) induced activation of NFκB is suppressed by α-lipoic acid in cultured endothelial cells.  Diabetes. 1997;  46 1481-1490
    CrossrefPubMedGoogle Scholar
  • 61 Bierhaus A, Hofmann MA, Ziegler R, Nawroth PP. AGEs and their interaction with AGE-receptors in vascular disease and diabetes mellitus I. The AGE concept.  Cardiovasc Res. 1998;  37 586-600
    CrossrefPubMedGoogle Scholar
  • 62 Haslbeck KM, Schleicher E, Bierhaus A, Nawroth P, Haslbeck M, Neundörfer B, Heuss D. The AGE/RAGE/NF-(kappa)B pathway may contribute to the pathogenesis of polyneuropathy in impaired glucose tolerance (IGT).  Exp Clin Endocrinol Diabetes. 2005;  113 288-291
    Article in Thieme ConnectPubMedGoogle Scholar
  • 63 Humpert PM, Djuric Z, Kopf S, Rudofsky G, Morcos M, Nawroth PP, Bierhaus A. Soluble RAGE but not endogenous secretory RAGE is associated with albuminuria in patients with type 2 diabetes.  Cardiovasc Diabetol. 2007;  6 9
    CrossrefPubMedGoogle Scholar
  • 64 Humpert PM, Kopf S, Djuric Z, Wendt T, Morcos M, Nawroth PP, Bierhaus A. Plasma sRAGE is independently associated with urinary albumin excretion in type 2 diabetes.  Diabetes Care. 2006;  29 1111-1113
    CrossrefPubMedGoogle Scholar
  • 65 Humpert PM, Papadopoulos G, Schaefer K, Djuric Z, Konrade I, Morcos M, Nawroth PP, Bierhaus A. sRAGE and esRAGE are not associated with peripheral or autonomic neuropathy in type 2 diabetes.  Horm Metab Res. 2007;  39 899-902
    Article in Thieme ConnectPubMedGoogle Scholar
  • 66 Jensen LJ, Østergaard J, Flyvbjerg A. AGE-RAGE and AGE Cross-link interaction: important players in the pathogenesis of diabetic kidney disease.  Horm Metab Res. 2005;  37 ((Suppl 1)) 26-34
    Article in Thieme ConnectPubMedGoogle Scholar
  • 67 Mezzano S, Aros C, Droguett A, Burgos ME, Ardiles L, Flores C, Schneider H, Ruiz-Ortega M, Egido J. NF-kappaB activation and overexpression of regulated genes in human diabetic nephropathy.  Nephrol Dial Transplant. 2004;  19 2505-2512
    CrossrefPubMedGoogle Scholar
  • 68 Rudofsky  Jr  G, Isermann B, Schilling T, Schiekofer S, Andrassy M, Schneider JG, Morcos M, Humpert PM, Sayed AA, Witte S, Renn W, Pfohl M, Hamann A, Nosikov V, Schleicher E, Häring HU, Rudofsky G, Ritz E, Nawroth PP, Bierhaus A. A 63bp deletion in the promoter of rage correlates with a decreased risk for nephropathy in patients with type 2 diabetes.  Exp Clin Endocrinol Diabetes. 2004;  12 135-141
    Article in Thieme ConnectPubMedGoogle Scholar
  • 69 Wendt T, Tanji N, Guo J, Hudson BI, Bierhaus A, Ramasamy R, Arnold B, Nawroth PP, Yan SF, D’Agati V, Schmidt AM. Glucose, glycation, and RAGE: implications for amplification of cellular dysfunction in diabetic nephropathy.  J Am Soc Nephrol. 2003;  14 1383-1395
    CrossrefPubMedGoogle Scholar
  • 70 Clemens A, Siegel E, Gallwitz B. Global risk management in type 2 diabetes: blood glucose, blood pressure, and lipids – update on the background of the current guidelines.  Exp Clin Endocrinol Diabetes. 2004;  112 493-503
    Article in Thieme ConnectPubMedGoogle Scholar

Correspondence

Dr. M. Morcos

Department of Internal Medicine 1 (Endocrinology, Metabolism and Clinical Chemistry) & Renal Department

University of Heidelberg

INF 410

69120 Heidelberg

Germany

Phone: +49/6221/56 86 14

Fax: +49/6221/56 68 48

Email: michael_morcos@med.uni-heidelberg.de