Our publications
2023
Gene-by-gene screen of the unknown proteins encoded on Plasmodium falciparum chromosome 3.
- Kimmel J, Schmitt M, Sinner A, Jansen PWTC, Mainye S, Ramón-Zamorano G, Toenhake CG, Wichers-Misterek JS, Cronshagen J, Sabitzki R, Mesén-Ramírez P, Behrens HM, Bártfai R, Spielmann T.
- Cell Syst. 2023 Jan 18;14(1):9-23.e7. doi: 10.1016/j.cels.2022.12.001.
Clearing of hemozoin crystals in malaria parasites enables whole-cell STED microscopy.
- Kehrer J, Pietsch E, Heinze J, Spielmann T, Frischknecht F.
- J Cell Sci. 2023 Jan 1;136(1):jcs260399. doi: 10.1242/jcs.260399. Epub 2023 Jan 9.
2022
PMRT1, a Plasmodium-Specific Parasite Plasma Membrane Transporter, Is Essential for Asexual and Sexual Blood Stage Development
- Wichers JS, Mesén-Ramírez P, Fuchs G, Yu-Strzelczyk J, Stäcker J, von Thien H, Alder A, Henshall I, Liffner B, Nagel G, Löw C, Wilson D, Spielmann T, Gao S, Gilberger TW, Bachmann A, Strauss J.
- mBio. 2022 Apr 26;13(2):e0062322. doi: 10.1128/mbio.00623-22. Epub 2022 Apr 11.
Proximity-dependent biotinylation approaches to study apicomplexan biology
- Kimmel J, Kehrer J, Frischknecht F, Spielmann T.
- Mol Microbiol. 2022 Mar;117(3):553-568. doi: 10.1111/mmi.14815. Epub 2021 Oct 22.
2021
Characterization of Apicomplexan Amino Acid Transporters (ApiATs) in the Malaria Parasite Plasmodium falciparum
- Wichers JS, van Gelder C, Fuchs G, Ruge JM, Pietsch E, Ferreira JL, Safavi S, von Thien H, Burda PC, Mesén-Ramirez P, Spielmann T, Strauss J, Gilberger TW, Bachmann A.
- mSphere. 2021 Dec 22;6(6):e0074321. doi: 10.1128/mSphere.00743-21. Epub 2021 Nov 10.
The parasitophorous vacuole nutrient channel is critical for drug access in malaria parasites and modulates the artemisinin resistance fitness cost
- Mesén-Ramírez P, Bergmann B, Elhabiri M, Zhu L, von Thien H, Castro-Peña C, Gilberger TW, Davioud-Charvet E, Bozdech Z, Bachmann A, Spielmann T.
- Cell Host Microbe. 2021 Dec 8;29(12):1774-1787.e9. doi: 10.1016/j.chom.2021.11.002. Epub 2021 Dec 3. Comment in Compensating P. falciparum artemisinin resistance. Platon L, Cao J, Ménard D. Cell Host Microbe. 2021 Dec 8;29(12):1732-1734. doi: 10.1016
The newly discovered role of endocytosis in artemisinin resistance
- Behrens HM, Schmidt S, Spielmann T.
- Med Res Rev. 2021 Nov;41(6):2998-3022. doi: 10.1002/med.21848. Epub 2021 Jul 26.
Identification of novel inner membrane complex and apical annuli proteins of the malaria parasite Plasmodium falciparum
- Wichers JS, Wunderlich J, Heincke D, Pazicky S, Strauss J, Schmitt M, Kimmel J, Wilcke L, Scharf S, von Thien H, Burda PC, Spielmann T, Löw C, Filarsky M, Bachmann A, Gilberger TW.
- Cell Microbiol. 2021 Sep;23(9):e13341. doi: 10.1111/cmi.13341. Epub 2021 May 16. PMID: 33830607
2020
The ZIP Code of Vesicle Trafficking in Apicomplexa: SEC1/Munc18 and SNARE Proteins
- Bisio H, Chaabene RB, Sabitzki R, Maco B, Marq JB, Gilberger TW, Spielmann T, Soldati-Favre D.
- mBio. 2020 Oct 20;11(5):e02092-20. doi: 10.1128/mBio.02092-20.
Introduction of Scaffold Nitrogen Atoms Renders Inhibitors of the Malarial l-Lactate Transporter, PfFNT, Effective against the Gly107Ser Resistance Mutation.
- Walloch P, Henke B, Häuer S, Bergmann B, Spielmann T, Beitz E.
- J Med Chem. 2020 Sep 10;63(17):9731-9741. doi: 10.1021/acs.jmedchem.0c00852. Epub PMID: 32816478
Endocytosis in Plasmodium and Toxoplasma Parasites
- Spielmann T, Gras S, Sabitzki R, Meissner M.
- Trends Parasitol.2020 Jun;36(6):520-532. doi: 10.1016/j.pt.2020.03.010. PMID: 32340866 Review.
A Kelch13-defined endocytosis pathway mediates artemisinin resistance in malaria parasites
- Birnbaum J*, Scharf S*, Schmidt S, Jonscher E, Hoeijmakers WAM, Flemming S, Toenhake CG, Schmitt M, Sabitzki R, Bergmann B, Fröhlke U, Mesen-Ramirez P, Blancke Soares A, Herrmann H, Bártfai R and Spielmann T
- Science, in press
2019
Epigenetic reader complexes of the human malaria parasite, Plasmodium falciparum.
- Hoeijmakers WAM, Miao J, Schmidt S, Toenhake CG, Shrestha S, Venhuizen J, Henderson R, Birnbaum J, Ghidelli-Disse S, Drewes G, Cui L, Stunnenberg HG, Spielmann T, Bártfai R.
- Nucleic Acids Res. 2019 Dec 16;47(22):11574-11588. doi: 10.1093/nar/gkz1044.
EXP1 is critical for nutrient uptake across the parasitophorous vacuole membrane of malaria parasites.
- Mesén-Ramírez P, Bergmann B, Tran TT, Garten M, Stäcker J, Naranjo-Prado I, Höhn K, Zimmerberg J, Spielmann T
- PLoS Biol. 2019 Sep 30;17(9):e3000473. doi: 10.1371/journal.pbio.3000473. eCollection 2019 Sep.
Delayed death in the malaria parasite Plasmodium falciparum is caused by disruption of prenylation-dependent intracellular trafficking.
- Kennedy K, Cobbold SA, Hanssen E, Birnbaum J, Spillman NJ, McHugh E, Brown H, Tilley L, Spielmann T, McConville MJ, Ralph SA.
- PLoS Biol. 2019 Jul 18;17(7):e3000376. doi: 10.1371/journal.pbio.3000376. eCollection 2019 Jul.
PfVPS45 Is Required for Host Cell Cytosol Uptake by Malaria Blood Stage Parasites.
- Jonscher E, Flemming S, Schmitt M, Sabitzki R, Reichard N, Birnbaum J, Bergmann B, Höhn K, Spielmann T.
- Cell Host Microbe. 2019 Jan 9;25(1):166-173.e5. doi: 10.1016/j.chom.2018.11.010. Epub 2018 Dec 20.
2018
Identification of novel parasitophorous vacuole proteins in P. falciparum parasites using BioID.
- Khosh-Naucke M, Becker J, Mesén-Ramírez P, Kiani P, Birnbaum J, Fröhlke U, Jonscher E, Schlüter H, Spielmann T
- Int J Med Microbiol. 2018 Jan;308(1):13-24. doi: 10.1016/j.ijmm.2017.07.007. Epub 2017 Jul 27.
2017
Substrate-analogous inhibitors exert antimalarial action by targeting the Plasmodium lactate transporter PfFNT at nanomolar scale.
- Golldack A, Henke B, Bergmann B, Wiechert M, Erler H, Blancke Soares A, Spielmann T, Beitz E
- PLoS Pathog. 2017 Feb 8;13(2):e1006172. doi: 10.1371/journal.ppat.1006172. eCollection 2017 Feb.
Progress in imaging methods: insights gained into Plasmodium biology.
- De Niz M, Burda PC, Kaiser G, Del Portillo HA, Spielmann T, Frischknecht F, Heussler VT.
- Nat Rev Microbiol. 2017 Jan;15(1):37-54. doi: 10.1038/nrmicro.2016.158. Epub 2016 Nov 28.
Birnbaum J, Flemming S, Reichard N, Soares AB, Mesén-Ramírez P, Jonscher E, Bergmann B, Spielmann T (2017) A genetic system to study Plasmodium falciparum protein function.
- Birnbaum J, Flemming S, Reichard N, Soares AB, Mesén-Ramírez P, Jonscher E, Bergmann B, Spielmann T
- Nat Methods 2017, 14(4):450-456. doi: 10.1038/nmeth.4223. PMID: 28288121.
2016
Stable Translocation Intermediates Jam Global Protein Export in Plasmodium falciparum Parasites and Link the PTEX Component EXP2 with Translocation Activity
- Mesén-Ramírez P, Reinsch F, Blancke Soares A, Bergmann B, Ullrich AK, Tenzer S, Spielmann T.
- PLoS Pathog. 2016;12(5):e1005618. doi: 10.1371/journal.ppat.1005618
Hierarchical phosphorylation of apical membrane antigen 1 is required for efficient red blood cell invasion by malaria parasites.
- Prinz B, Harvey KL, Wilcke L, Ruch U, Engelberg K, Biller L, Lucet I, Erkelenz S, Heincke D, Spielmann T, Doerig C, Kunick C, Crabb BS, Gilson PR, Gilberger TW.
- Sci Rep. 2016 Oct 4;6:34479. doi: 10.1038/srep34479.
The machinery underlying malaria parasite virulence is conserved between rodent and human malaria parasites
- De Niz M, Ullrich AK, Heiber A, Blancke Soares A, Pick C, Lyck R, Keller D, Kaiser G, Prado M, Flemming S, Del Portillo H, Janse CJ, Heussler V, Spielmann T.
- Nat Commun. 2016;7:11659. doi: 10.1038/ncomms11659
Boddey JA, O'Neill MT, Lopaticki S, Carvalho TG, Hodder AN, Nebl T, Wawra S, van West P, Ebrahimzadeh Z, Richard D, Flemming S, Spielmann T, Przyborski J, Babon JJ, Cowman AF
- Export of malaria proteins requires co-translational processing of the PEXEL motif independent of phosphatidylinositol-3-phosphate binding
- Nat Commun. 2016, 7:10470. doi: 10.1038/ncomms10470
Pellicle formation in the malaria parasite
- Kono M, Heincke D, Wilcke L, Wong T, Bruns C, Herrmann S, Spielmann T, Gilberger TW
- J Cell Sci. 2016, 129(4):673-80. doi: 10.1242/jcs.181230
2015
Critical Steps in Protein Export of Plasmodium falciparum Blood Stages
- Spielmann T, Gilberger TW
- Trends Parasitol. 2015, 31(10):514-25. doi: 10.1016/j.pt.2015.06.010
The Plasmodium falciparum exportome contains non-canonical PEXEL/HT proteins
- Schulze J, Kwiatkowski M, Borner J, Schlüter H, Bruchhaus I, Burmester T, Spielmann T#, Pick C# (#, equal contributors)
- Mol Microbiol. 2015, 97:301-14. doi: 10.1111/mmi.13024
Identity of a Plasmodium lactate/H(+) symporter structurally unrelated to human transporters
- Wu, B., Rambow, J., Bock, S., Holm-Bertelsen, J., Wiechert, M., Soares, A.B., Spielmann, T., Beitz, E.
- Nat. Commun. 2015, 6:6284. doi: 10.1038/ncomms7284
The role of palmitoylation for protein recruitment to the inner membrane complex of the malaria parasite
- Wetzel, J., Herrmann, S., Swapna, L.S., Prusty, D., Peter, A.T., Kono, M., Saini, S., Nellimarla, S., Wong, T.W., Wilcke, L., Ramsay, O., Cabrera, A., Biller, L., Heincke, D., Mossman, K., Spielmann, T., Ungermann, C., Parkinson, J., Gilberger, T.W
- J. Biol. Chem. 2015, 290:1712-28. doi: 10.1074/jbc.M114.598094
2013
Protein export in malaria parasites: many membranes to cross.
- Marti, M., and Spielmann, T.
- Curr Opin Microbiol. 2013 Aug;16(4):445-51.
Specific phosphorylation of the PfRh2b invasion ligand of Plasmodium falciparum.
- Engelberg, K., Paul, A.S., Prinz, B., Kono, M., Ching, W., Heincke, D., Dobner, T., Spielmann, T., Duraisingh, M., and Gilberger, T.-W.
- Biochem J. 2013 Jun 15;452(3):457-66.
Identification of New PNEPs Indicates a Substantial Non-PEXEL Exportome and Underpins Common Features in Plasmodium falciparum Protein Export
- Heiber, A., Kruse, F., Pick, C., Grüring, C., Flemming, S., Oberli, A., Schoeler, H., Retzlaff, S., Mesén-Ramírez, P., Hiss, J.A., Kadekoppala, M., Hecht, L., Holder, A.H., Gilberger, T.-W., Spielmann, T.
- PLoS Pathog. 2013;9(8):e1003546. doi: 10.1371/journal.ppat.1003546. Epub 2013 Aug 8.
2012
Wherever I may roam: Protein and membrane trafficking in P. falciparum-infected red blood cells.
- Deponte, M., Hoppe, H.C., Lee, M, . Maier, A.G., Richard, D., Rug, M., Spielmann, T, and Przyborski, J.M.
- Mol Biochem Parasitol. 2012 Dec;186(2):95-116.
Dissection of minimal sequence requirements for rhoptry membrane targeting in the malaria parasite.
- Cabrera, A., Herrmann, S., Warszta, D., Santos, J.M., John Peter, A.T., Kono, M., Debrouver, S., Jacobs, T., Spielmann, T., Ungermann, C., Soldati-Favre, D., and Gilberger T.W.
- Traffic. 2012 Oct;13(10):1335-50.
Molecular make-up of the Plasmodium parasitophorous vacuolar membrane.
- Spielmann, T., Montagna, G., Hecht, L., and Matuschewski, K.
- Int J Med Microbiol. 2012 Oct;302(4-5):179-86.
Evolution and Architecture of the Inner Membrane Complex in Asexual and Sexual Stages of the Malaria Parasite.
- Kono, M., Herrmann, S., Loughran, N.B., Cabrera, A., Engelberg, K., Lehmann, C., Sinha, D., Prinz, B., Ruch, U., Heussler, V., Spielmann, T., Parkinson, J., and Gilberger, T.W.
- Mol Biol Evol. 2012 Sep;29(9):2113-32.
Host cell deformability is linked to transmission in the human malaria parasite Plasmodium falciparum.
- Aingaran, M., Zhang, R., Law, S.K.Y., Peng, Z., Undisz, A., Meyer, E., Diez-Silva, M., Burke, T.A., Spielmann, T., Lim, C.T., Suresh, S., Dao, M., and Marti, M.
- Cell Microbiol. 2012 Jul;14(7):983-93.
Uncovering common principles in protein export of malaria parasites.
- Heiber, A., Kruse, F., Pick, C., Grüring, C., Flemming, S., Oberli, A., Schoeler, H., Retzlaff, S., Mesén-Ramírez, P., Hiss, J.A., Kadekoppala, M., Hecht, L., Holder, A.H., Grüring, C., Heiber, A., Kruse, F., Flemming, S., Franci, G., Colombo, S., Fasana, F., Schöler, H., Borgese, N., Stunnenberg, H.G., Przyborski, J.M., Gilberger, T.-W., and Spielmann, T.
- Cell Host Microbe. 2012 Nov 15;12(5):717-29.
Imaging of live malaria blood stage parasites.
- Grüring, C. and Spielmann, T. (2012)
- Methods Enzymol. 2012;506:81-92.
2011
Phylogenomic analyses of malaria parasites and evolution of their exported proteins.
- Pick, C., Ebersberger, I., Spielmann, T., Bruchhaus, I., and Thorsten Burmester, T. (2011) BMC Evol. Biol., 11:167 (doi:10.1186/1471-2148-11-167)
- BMC Evol Biol. 2011 Jun 15;11:167. doi: 10.1186/1471-2148-11-167.
Development and Host Cell Modifications of Plasmodium falciparum Blood Stages in 4D.
- Grüring, C., Heiber, A., Kruse, K., Ungefehr, J., Gilberger, T.-W., and Spielmann, T.
- Nat Commun. 2011 Jan 25;2:165.
2010
Transcriptional profiling of growth perturbations of the human malaria parasite Plasmodium falciparum.
- Hu, G., Cabrera, A., Kono, M., Mok, S., Chaal, B.K., Haase, S., Engelberg, K., Cheemadan, S., Spielmann, T., Preiser, P.R., Gilberger, T.W., Bozdech, Z.
- Nat Biotechnol. 2010 Jan;28(1):91-8. doi: 10.1038/nbt.1597. Epub 2009 Dec 27.
Protein export in malaria parasites: do multiple export motifs add up to multiple export pathways?
- Spielmann, T., & Gilberger T.W.
- Trends Parasitol. 2010 Jan;26(1):6-10. doi: 10.1016/j.pt.2009.10.001. Epub 2009 Oct 29.
2009
A novel Plasmodium falciparum erythrocyte binding protein associated with the merozoite surface, PfDBLMSP.
- Wickramarachchi, T., Cabrera, A.L., Sinha, D., Dhawan, S., Chandran, T., Devi, Y.S., Kono, M., Spielmann, T., Gilberger, T.W., Chauhan, V.S., Mohmmed, A.
- Int J Parasitol. 2009 Jun;39(7):763-73.
Functional analysis of the leading malaria vaccine candidate AMA-1 reveals an essential role for the cytoplasmic domain in the invasion process.
- Treeck, M., Zacherl, S., Herrmann, S., Cabrera, A., Kono, M., Struck, N.S., Engelberg, K., Haase, S., Frischknecht, F., Miura, K., Spielmann, T., Gilberger, T.W.
- PLoS Pathog. 2009 Mar;5(3):e1000322. doi: 10.1371/journal.ppat.1000322. Epub 2009 Mar 6.
Sequence requirements for the export of the P. falciparum Maurer's clefts protein REX2.
- Haase, S., Herrmann, S., Grüring, C., Heiber, A., Jansen, P.W., Langer, C., Treeck, M., Cabrera, L. A., Bruns, C., Struck, N., Kono, M., Engelberg, K., Ruch, U., Stunnenberg, H.G., Gilberger, T.W., and Spielmann, T.
- Mol Microbiol. 2009 Feb;71(4):1003-17. doi: 10.1111/j.1365-2958.2008.06582.x. Epub 2008 Dec 18.
2008
Reliable transfection of Plasmodium falciparum using non-commercial plasmid mini preparations.
- Spielmann, T., Dixon, M.W.A., Hernandez-Valladares, M., Hannemann, M., Trenholme, K.R, and Gardiner, D.L.
- Int J Parasitol. 2006 Oct;36(12):1245-8. Epub 2006 Jul 28.
Targeting of the Ring Exported Protein 1 to the Maurer's clefts is mediated by a two phase process.
- Dixon, M.W.A., Hawthorne, P.L., Spielmann, T., Anderson, K.L., Trenholme, K.R., and Gardiner, D.L.
- Traffic. 2008 Aug;9(8):1316-26. doi: 10.1111/j.1600-0854.2008.00768.x. Epub 2008 May 19.
Targeted mutagenesis of the ring exported protein-1 of Plasmodium falciparum disrupts the architecture of Maurer's cleft organelles.
- Hanssen, E., Hawthorne, P.L., Dixon, M.W.A., Trenholme, K.R., McMillan, P., Spielmann, T., Gardiner, D.L., and Tilley, L.
- Mol Microbiol. 2008 Aug;69(4):938-53. doi: 10.1111/j.1365-2958.2008.06329.x. Epub 2008 Jun 28.
Plasmodium falciparum possesses two GRASP proteins that are differentially targeted to the golgi apparatus via a higher and lower eukaryote-like mechanism.
- Struck, N., Herrmann, S., Langer, C., Krueger, K., Foth, B., Engelberg, K., Cabrera, L. A., Haase, S., Treeck, M., Marti, M., Cowman, A.F., Spielmann, T., and Gilberger, T.W. (2008) Plasmodium falciparum possesses two GRASP proteins that are differentially targeted to the golgi apparatus via a higher and lower eukaryote-like mechanism. J. Cell Sci., 121:2123-9.
- J Cell Sci. 2008 Jul 1;121(Pt 13):2123-9. doi: 10.1242/jcs.021154. Epub 2008 Jun 3.
Spatial dissection of the cis- and trans-Golgi compartments in the malaria parasite Plasmodium falciparum.
- Struck, N., Herrmann, S., Schmuck-Barkmann, I., Diaz, S., Haase, S., Cabrera, A., Treeck, M., Bruns, C., Langer, C., Cowman, A.F., Marti, M., Spielmann, T., and Gilberger, T.W. (2008) Mol. Microbiol., 67:1320-30.
- Mol Microbiol. 2008 Mar;67(6):1320-30. doi: 10.1111/j.1365-2958.2008.06125.x. Epub 2008 Feb 12.
Characterization of a conserved rhoptry-associated leucine zipper-like protein in the malaria parasite Plasmodium falciparum.
- Haase, S., Cabrera, A., Langer, C., Treeck, M., Struck, N., Herrmann, S., Jansen, P.W., Bruchhaus, I., Bachmann, A., Diaz, S., Cowman, A.F., Stunnenberg, H.G., Spielmann, T., and Gilberger, T.W.
- Infect Immun. 2008 Mar;76(3):879-87. doi: 10.1128/IAI.00144-07. Epub 2008 Jan 3.
2007
New therapeutic approaches for malaria: molecular aspects of erythrocyte invasion.
- Spielmann, T., and Gilberger T.-W. (2007) New therapeutic approaches for malaria: molecular aspects of erythrocyte invasion.
- Dtsch Med Wochenschr. 2007 Nov;132(45):2383-6.
The M18 Aspartyl Aminopeptidase of the human malaria parasite Plasmodium falciparum.
- Teuscher, F., Lowther, J., Skinner-Adams, T.S., Spielmann, T., Dixon, M.W.A., Stack, C.M., Donnelly, S., Mucha, A., Kafarski, P., Vassiliou, S., Gardiner, D.L., Dalton, J.P., and Trenholme, K.R.
- J Biol Chem. 2007 Oct 19;282(42):30817-26. Epub 2007 Aug 20.
Selective permeabilization of the host cell membrane of Plasmodium falciparum-infected red blood cells with streptolysin O and equinatoxin II.
- Spielmann, T., Jackson, K.E.*, Hanssen, E., Adisa, A., Separovic, F., Dixon, M.W., Trenholme, K.R., Hawthorne, P.L., Gardiner, D.L., Gilberger, T., and Tilley, L. *joint first authors
- Biochem J. 2007 Apr 1;403(1):167-75.
2006
A cluster of ring stage-specific genes linked to a locus implicated in cytoadherence in Plasmodium falciparum codes for PEXEL negative and PEXEL positive proteins exported into the host cell.
- Spielmann, T., Hawthorne, P.L., Dixon, M.W.A., Hannemann, M., Klotz, K., Kemp, D.J., Klonis, N., Tilley, L., Trenholme K.R., and Gardiner, D.L.
- Mol Biol Cell. 2006 Aug;17(8):3613-24. Epub 2006 Jun 7.
Organisation of ETRAMPs and EXP-1 at the parasite-host cell interface of malaria parasites.
- Spielmann, T., Gardiner, D.L., Beck, H.-P., Trenholme, K.R., Kemp, D.J.
- Mol Microbiol. 2006 Feb;59(3):779-94.
2004
A Novel Plasmodium falciparum Ring Stage Protein, REX, is Located in Maurer's Clefts.
- Hawthorne, P.L., Trenholme, K.R., Skinner-Adams, T.S., Spielmann, T., Fischer, K., Dixon, M.W.A., Ortega, M.R., Anderson, K.L., Kemp, D.J., Gardiner, D.L.
- Mol Biochem Parasitol. 2004 Aug;136(2):181-9.
2003
MAHRP-1, a novel Plasmodium falciparum histidine-rich protein, binds ferriprotoporphyrin IX and localises to the Maurer's clefts.
- Spycher, C., Klonis, N., Spielmann, T., Kump, E., Steiger, S., Tilley, L., Beck, H.-P.
- J Biol Chem. 2003 Sep 12;278(37):35373-83. Epub 2003 Jun 18.
etramps, a new Plasmodium falciparum gene family coding for developmentally regulated and highly charged membrane proteins located at the parasite-host cell interface.
- Spielmann, T., Fergusen, D.J.P., Beck, H.-P.
- Mol Biol Cell. 2003 Apr;14(4):1529-44.
2000
Analysis of stage-specific transcription in Plasmodium falciparum reveals a set of genes exclusively transcribed in ring stage parasites.
- Spielmann, T., Beck, H.-P.
- Mol Biochem Parasitol. 2000 Dec;111(2):453-8.