Allogeneic stem cell transplantation for elderly patients with myelodysplastic syndrome

Blood

Volumn 119, Issue 24, 2012, Pages 5632-5639

  Kröger, Nicolaus ^a  

^a UNIVERSITY MEDICAL CENTER HAMBURG EPPENDORF   (Germany)

Author keywords

[No Author keywords available]

Indexed keywords

5 AZA 2' DEOXYCYTIDINE; ANTHRACYCLINE; ARABINOSIDE; AZACITIDINE; CLOFARABINE; CYCLOPHOSPHAMIDE; CYTARABINE; CYTOSINE; FLUDARABINE; TREOSULFAN;

ACUTE GRANULOCYTIC LEUKEMIA; AGED; ALLOGENEIC STEM CELL TRANSPLANTATION; BLAST CELL; CLASSIFICATION ALGORITHM; COMORBIDITY; DISEASE FREE SURVIVAL; DONOR LYMPHOCYTE INFUSION; DONOR SELECTION; DRUG SAFETY; GERIATRIC PATIENT; GRAFT RECIPIENT; HEMATOPOIETIC CELL TRANSPLANTATION COMORBIDITY INDEX; HUMAN; INTERNATIONAL PROGNOSTIC SCORING SYSTEM; IRON OVERLOAD; MOLECULAR GENETICS; MORTALITY; MYELOABLATIVE CONDITIONING; MYELODYSPLASTIC SYNDROME; NONMYELOABLATIVE CONDITIONING; OVERALL SURVIVAL; PERSONALIZED MEDICINE; PRIORITY JOURNAL; RATING SCALE; RECOMMENDED DRUG DOSE; RECURRENCE RISK; REDUCED INTENSITY CONDITIONING; RELAPSE; REMISSION; REVIEW; RISK ASSESSMENT; RISK REDUCTION; TREATMENT INDICATION; WHOLE BODY RADIATION;

EID: 84862486519     PISSN: 00064971     EISSN: 15280020     Source Type: Journal    
DOI: 10.1182/blood-2011-12-380162     Document Type: Review

References (86)

1. Greenberg P, Cox C, LeBeau MM, et al. International scoring system for evaluating prognosis in myelodysplastic syndromes. Blood. 1997;89(6):2079-2088. (Pubitemid 27132124)
2. Nimer S. Myelodysplastic syndromes. Blood. 2008;111(10):4841-4851.
3. Swerdlow SH, Campo E, Harris NL, et al. WHO Classification of Tumours of Haematopoietic and Lymphoid Tissues. Lyon, France: IARC Press; 2008.
4. Ma X, Does M, Raza A, et al. Myelodysplastic syndromes: incidence and survival in the United States. Cancer. 2007;109(8):1536-1542. (Pubitemid 46595689)
5. Oliansky DM, Antin JH, Bennett JM, et al. The role of cytotoxic therapy with hematopoietic stem cell transplantation in the therapy of myelodysplastic syndromes: an evidence-based review. Biol Blood Marrow Transplant. 2009;15(2):137-172.
6. Appelbaum FR, Storb R, Ramberg RE, et al. Treatment of preleukemia syndromes with marrow transplantation. Blood. 1987;69(1):92-96. (Pubitemid 17007080)
7. de Witte T, Hermans J, Vossen J, et al. Haematopoietic stem cell transplantation for patients with myelodysplastic syndromes and secondary acute myeloid leukemia: a report on behalf of the Chronic Leukemia Working Party of the European Group for Blood and Marrow Transplantation (EBMT). Br J Haematol. 2000;110(3):620-630.
8. Sierra J, Peaposrez WS, Rozman C, et al. Bone marrow transplantation from HLA-identical siblings as treatment for myelodysplasia. Blood. 2002;100(6):1997-2004. (Pubitemid 35001229)
9. Sutton L, Chastang C, Ribaud P, et al. Factors influencing outcome in de-novo myelodysplastic syndromes treated by allogeneic bone marrow transplantation: a long term study of 71 patients. Societé Francaise de Greffe de Moelle. Blood. 1996;88(1):358-365. (Pubitemid 26230708)
10. Anderson J, Anasetti C, Appelbaum F, et al. Unrelated donor bone marrow transplantation for myelodysplasia (MDS) and MDS-related acute myeloid leukemia. Br J Haematol. 1996;93(1):59-67. (Pubitemid 26130716)
11. Oostervald M, Suciu S, Verhoef G, et al. The presence of an HLA-identical sibling donor has no impact on outcomes of patients with high risk MDS or secondary AML (sAML) treated with intensive chemotherapy followed by transplantation: results of a prospective study of the EORTC, EBMT, SAKK, and GIMEMA Leukemia groups (EORTC 06921). Leukemia. 2003;17(5):859-868. (Pubitemid 36626312)
12. Deeg HJ, Shulman HM, Anderson JE, et al. Allogeneic and syngeneic marrow transplantation for myelodysplastic syndrome in patients 55 to 65 years of age. Blood. 2000;95(4):1188-1194. (Pubitemid 30099825)
13. Slavin S, Nagler A, Naperstek E, et al. Nonmyeloablative stem cell transplantation and cell therapy as an alternative to conventional bone marrow transplantation with lethal cytoreduction for the treatment of malignant and non-malignant hematologic diseases. Blood. 1998;91(3):756-763. (Pubitemid 28078275)
14. Giralt S, Estey E, Albitar M, et al. Engraftment of allogeneic hematopoietic progenitor cells with purine analog containing chemotherapy: harnessing graft-versus-leukemia without myeloablative therapy. Blood. 1997;89(12):4531-4536. (Pubitemid 27260263)
15. Laport GG, Sandmaier BM, Storer BE, et al. Reduced-intensity conditioning followed by allogeneic hematopoietic cell transplantation for adult patients with myelodysplastic syndrome and myeloproliferative disorders. Biol Blood Marrow Transplant. 2008;14(2):246-255.
16. Deeg HJ, Storer B, Slattery JT, et al. Conditioning with targeted busulfan and cyclophosphamide for hemopoietic stem cell transplantation from related and unrelated donors in patients with myelodysplastic syndrome. Blood. 2002;100(4):1201-1207. (Pubitemid 34864272)
17. Lim Z, Brand R, Martino R, et al. Allogeneic hematopoietic stem cell transplantation for patients 50 years or older with myelodysplastic syndromes or secondary acute myeloid leukemia. J Clin Oncol. 2010;28(3):405-411.
18. McClune BL, Weisdorf DJ, Pedersen TL, et al. Effect of age on outcome of reduced-intensity hematopoietic cell transplantation for older patients with acute myeloid leukemia in first complete remission or with myelodysplastic syndrome. J Clin Oncol. 2010;28(11):1878-1887.
19. Weisdorf D, Hakke R, Blazar B, et al. Risk factors for acute graft-versus-host disease in histocompatible donor bone marrow transplantation. Transplantation. 1991;51(6):1197-1203.
20. Kollmann C, Howe CW, Anasetti C, et al. Donor characteristics as risk factors in recipients after transplantation of bone marrow from unrelated donors: the effect of donor age. Blood. 2001;98(7):2043-2051.
21. Zipperer E, Pelz D, Nachtkamp K, et al. The hematopoietic stem cell transplantation comorbidity index is of prognostic relevance for patients with myelodysplastic syndrome. Haematologica. 2009;94(5):729-732.
22. Sorror ML, Sandmaier BM, Storer BE, et al. Comorbidity and disease status based risk stratification of outcomes among patients with acute myeloid leukemia or myelodysplasia receiving allogeneic hematopoietic cell transplantation. J Clin Oncol. 2007;25(27):4246-4254. (Pubitemid 47554249)
23. Lim ZY, Ingram W, Brand R, et al. Impact of pretransplant comorbidities on alemtuzumab-based reduced-intensity conditioning allogeneic hematopoietic SCT for patients with high-risk myelodysplastic syndrome and AML. Bone Marrow Transplant. 2010;45(4):633-639.
24. Boehm A, Sperr WR, Leitner G, et al. Comorbidity predicts survival in myelodysplastic syndromes or secondary acute myeloid leukemia after allogeneic stem cell transplantation. Eur J Clin Invest. 2008;38(12):945-952.
25. Cortelezzi A, Cattaneo C, Christiani S, et al. Non-transferrin-bound iron in myelodysplastic syndromes: a marker of ineffective erythropoiesis? Hematol J. 2000;1(3):153-158.
26. Pullarkat V. Objectives of iron chelation therapy in myelodysplastic syndromes: more than meets the eye? Blood. 2009;114(26):5251-5255.
27. Armand P, Kim HT, Cutler CS, et al. Prognostic impact of elevated pretransplantation serum ferritinin patients undergoing myeloablative stem cell transplantation. Blood. 2007;109(10):4586-4588. (Pubitemid 46743431)
28. Pullarkat V, Blanchard S, Tegtmeier B, et al. Iron overload adversely affects outcome of allogeneic hematopoietic cell transplantation. Bone Marrow Transplant. 2008;42(12):799-805.
29. Alessandrino EP, Della Porta MG, Bacigalupo A, et al. Prognostic impact of pre-transplantation transfusion history and secondary iron overload in patients with myelodysplastic syndrome undergoing allogeneic stem cell transplantation: a GITMO study. Haematologica. 2010;95(3):476-484.
30. Armand P, Kim HT, Rhodes J, et al. Iron overload in patients with acute leukemia or MDS undergoing myeloablative stem cell transplantation. Biol Blood Marrow Transplant. 2011;17(6):852-860.
31. Durken M, Nielsen P, Knobel S, et al. Nontransferrinbound iron in serum of patients receiving bone marrow transplants. Free Radic Biol Med. 1997;22(7):1159-1163. (Pubitemid 27171957)
32. Fritsch A, Langebrake C, Nielsen P, et al. Deferasirox (Exjade) given during conditioning regimen (FLAMSA/Busulfan/ATG) reduces the appearance of labile plasma iron in patients undergoing allogeneic stem cell transplantation [abstract]. Blood (ASH Annual Meeting Abstracts). 2011: Abstract 3023.
33. Sahlstedt L, von Bornsdorf L, Ebeling F, et al. Effective binding of free iron by a single intravenous dose of human apotransferrin in haematological stem cell transplant patients. Br J Haematol. 2002;119(2):547-553. (Pubitemid 35365512)
34. Martino R, Iacobelli S, Brand R, et al. Myelodysplastic Syndrome Subcommittee of the Chronic Leukemia Working Party of the European Blood and Marrow Transplantation Group: retrospective comparison of reduced-intensity conditioning and conventional high-dose conditioning for allogeneic hematopoietic stem cell transplantation using HLA-identical sibling donors in myelodysplastic syndromes. Blood. 2006;108(3):836-846. (Pubitemid 44154615)
35. Luger S, Ringden O, Zhang MJ, et al. Similar outcomes using myeloablative vs reduced-intensity allogeneic transplant preparative regimens for AML or MDS. Bone Marrow Transplant. 2012;47(2):203-211.
36. de Lima M, Anagnostopoulos A, Munsell M, et al. Nonablative versus reduced-intensity conditioning regimens in the treatment of acute myeloid leukemia and high-risk myelodysplastic syndrome: dose is relevant for long-term disease control after allogeneic hematopoietic stem cell transplantation. Blood. 2004;104(3):865-872. (Pubitemid 38970586)
37. Shimoni A, Hardan I, Shem-Tov N, et al. Allogeneic hematopoietic stem cell transplantation in AML and MDS using myeloablative versus re-duced- intensity conditioning: the role of dose intensity. Leukemia. 2006;20(2):322-328. (Pubitemid 43148672)
38. Andersson BS, Kashyap A, Gian V, et al. Conditioning therapy with intravenous busulfan and cyclophosphamide (IV BuCy2) for hematologic malignancies prior to allogeneic stem cell transplantation: a phase II study. Biol Blood Marrow Transplant. 2002;8(3):145-154. (Pubitemid 34285073)
39. Russell JA, Tran HAT, Quinlan D, et al. Once daily intravenous busulfan given with fludarabine as conditioning for allogeneic stem cell transplantation study of pharmacokinetics and early clinical outcomes. Biol Blood Marrow Transplant. 2002;8(9):468-476. (Pubitemid 35174820)
40. de Lima M, Couriel D, Thall PF, et al. Once daily intravenous busulfan and fludarabine: clinical and pharmacokinetic results of a myeloablative reducedtoxicity conditioning regimen for allogeneic stem cell in AML and MDS. Blood. 2004;104(3):857-864. (Pubitemid 38970585)
41. Ruutu T, Volin L, Beelen DW, et al. Reducedtoxicity conditioning with treosulfan and fludarabine in allogeneic hematopoietic stem cell transplantation for myelodysplastic syndromes: final results of an international prospective phase II trial. Haematologica. 2011;96(9):1344-1350.
42. Kröger N, Shimoni A, Zabelina T, et al. Reducedtoxicity conditioning with treosulfan, fludarabine, and ATG as preparative regimen for allogeneic stem cell transplantation (alloSCT) in elderly patients with secondary acute myeloid leukemia (sAML) or myelodysplastic syndrome (MDS). Bone Marrow Transplant. 2006;37(4):339-344.
43. Nevill TJ, Shepherd JD, Sutherland HJ, et al. IPSS poor-risk karyotype as a predictor of outcome for patients with myelodysplastic syndrome following myeloablative stem cell transplantation. Biol Blood Marrow Transplant. 2009;15(2):205-213.
44. Onida F, Brand R, van Biezen A, et al. Impact of cytogenetics on outcome of patients with MDS or secondary AML undergoing allogeneic HSCT from HLA-identical siblings: a retrospective analysis of the EBMT-CLWP [abstract]. Blood (ASH Annual Meeting Abstracts). 2006;108: Abstract 2653.
45. Van Gelder M, Schetelig J, Volin Liisa, et al. Monosomal karyotype predicts poor outcome for MDS/sAML patients with chromosome 7 abnormalities after allogeneic stem cell transplantation for MDS/sAML: a study of the MDS subcommittee of the Chronic Leukemia Working Party of the European Group for Blood and Marrow Transplantation (EBMT) [abstract]. Blood (ASH Annual Meeting Abstracts). 2009: Abstract 293.
46. Deeg HJ, Scott BL, Appelbaum FR, Gooley T. Impact of novel 5-group cytogenetic risk classification of MDS on outcome after allogeneic hematopoietic cell transplantation (HCT) [abstract]. Blood (ASH Annual Meeting Abstracts). 2011;118: Abstract 666.
47. Platzbecker U, Mohr B, von Bonin M, et al. Lenalidomide as induction therapy before allogenic stem cell transplantation in a patient with proliferative CMML-2 and del(5q) not involving ERG1 locus. Leukemia. 2007;21:2384-2385.
48. Rüter B, Wijermans P, Claus R, et al. Preferential cytogenetic response to continuous intravenous low-dose decitabine (DAC) administration in myelodysplastic syndrome with monosomy 7. Blood. 2007;110:1080-1082.
49. Bejar R, Stevenson K, Abdel-Wahab O, et al. Clinical effect of point mutations in myelodysplastic syndromes. N Engl J Med. 2011;364(26):2496-2506.
50. Yoshida K, Sanada M, Shiraishi Y, et al. Frequent pathway mutations of splicing machinery in myelodysplasia. Nature. 2011;478:64-69.
51. Malcovati L, Papaemmanuil E, Bowen DT, et al. Clinical significance of SF3B mutations in myelodysplastic syndromes and myelodysplastic/ myeloproliferative neoplasms. Blood. 2011;118:6239-6246.
52. Nakai K, Kanda Y, Fukuhara S, et al. Value of chemotherapy before allogeneic hematopoietic stem cell transplantation from HLA-identical sibling donor for myelodysplastic syndrome. Leukemia. 2005;19(3):396-401. (Pubitemid 40403316)
53. Scott BL, Storer B, Loken MR, et al. Pretransplantation induction chemotherapy and post-transplantation relapse in patients with advanced myelodysplastic syndrome. Biol Blood Marrow Transplant. 2005;11(1):65-73. (Pubitemid 40057995)
54. Oran B, Giralt S, Saliba R, et al. Allogeneic hematopoietic stem cell transplantation for the treatment of high-risk myelogenous leukemia and myelodysplastic syndrome using reduced-intensity conditioning with fludarabine and melphalan. Biol Blood Marrow Transplant. 2007;13(4):454-462. (Pubitemid 46421890)
55. Schmid C, Schleuning M, Ledderose G, et al. Sequential regimen of chemotherapy, reducedintensity conditioning for allogeneic stem cell transplantation, and prophylactic donor lymphocyte transfusion in high-risk acute myeloid leukemia and myelodysplastic syndrome. J Clin Oncol. 2005;23(24):5675-5687.
56. Kröger N, Zabelina T, Wolschke C, et al. Induction chemotherapy followed immediately by busulfanbased reduced conditioning and allografting in elderly patients with advanced MDS or sAML [abstract]. Blood (ASH Annual Meeting Abstracts). 2009;114: Abstract 3387.
57. Buchholz S, Dammann E, Stadler M, et al. Cytoreductive treatment with clofarabine/ara-C combined with reduced-intensity conditioning and allogeneic stem cell transplantation in patients with high-risk, relapsed, or refractory acute myeloid leukemia and advanced myelodysplastic syndrome. Eur J Haematol. 2012;88(1):52-60.
58. Silverman LR, Demakos EP, Peterson BL, et al. Randomized controlled trial of azacitidine in patients with myelodysplastic syndrome: a study of the Cancer and Leukemia Group B. J Clin Oncol. 2002;20(10):2429-2440. (Pubitemid 34525728)
59. Fenaux P, Mufti GJ, Hellstrom-Lindberg E, et al. Efficacy of azacitidine compared with that of conventional care regimens in the treatment of higher-risk myelodysplastic syndromes: a randomized, open-label, phase III study. Lancet Oncol. 2009;10(3):223-232.
60. de Padua SL, de Lima M, Kantarjian H, et al. Feasibility of allo SCT after hypomethylating therapy with decitabine for myelodysplastic syndrome. Bone Marrow Transplant. 2009;43(11):839-843.
61. Field T, Perkins J, Huang Y, et al. 5-Azacitidine for myelodysplasia before allogeneic hematopoietic cell transplantation. Bone Marrow Transplant. 2010;45(2):255-260.
62. Malcovati L, Porta MG, Pascutto C, et al. Prognostic factors and life expectancy in myelodysplastic syndromes classified according to WHO criteria: a basis for clinical decision making. J Clin Oncol. 2005;23(30):7594-7603. (Pubitemid 46291823)
63. Malcovati L, Germing U, Kuendgen A, et al. Timedependent prognostic scoring system for predicting survival and leukemic evolution in myelodysplastic syndromes. J Clin Oncol. 2007;25(23):3503-3510. (Pubitemid 47315575)
64. Alessandrino EP, Della Porta MG, Bacigalupo A, et al. WHO classification and WPSS predict posttransplantation outcome in patients with myelodysplastic syndrome: a study from the Gruppo Italiano Trapianto di Modollo Osseo (GITMO). Blood. 2008;112(3):895-902.
65. Appelbaum FR, Anderson J. Allogeneic bone marrow transplantation for myelodysplastic syndromes: outcomes analysis according to IPSS scores. Leukemia. 1998;12(suppl 1):S25-S29. (Pubitemid 28479000)
66. de Witte T, Brand R, van Biezen A, et al. Allogeneic stem cell transplantation for patients with refractory anaemia with matched related and unrelated donors: delay of the transplant is associated with inferior survival. Br J Haematol. 2009;146(6):627-636.
67. Cutler CS, Lee SJ, Greenberg P, et al. A decision analysis of allogeneic bone marrow transplantation for the myelodysplastic syndromes: delayed transplantation for low-risk myelodysplasia is associated with improved outcome. Blood. 2004;104(2):579-585. (Pubitemid 38900045)
68. Kröger N, Putter H, Brand R, et al. Comparison of allogeneic stem cell transplantation and best supportive care in elderly patients with advanced MDS [abstract]. Blood (ASH Annual Meeting Abstracts). 2009;114: Abstract 3363.
69. Prébet T, Gore SD, Esterni B, et al. Outcome of high-risk myelodysplastic syndrome after azacitidine treatment failure. J Clin Oncol. 2011;29(24):3322-3327.
70. Arnold R, de Witte T, van Biezen A, et al. Unrelated bone marrow transplantation in patients with myelodysplastic syndromes and secondary acute myeloid leukemia: an EBMT survey. Bone Marrow Transplant. 1998;21(12):1213-1216. (Pubitemid 28318748)
71. Castro-Malaspina H, Harris RE, Gajewski J, et al. Unrelated donor marrow transplantation for myelodysplastic syndromes: outcome analysis in 510 transplants facilitated by the National Marrow Donor Program. Blood. 2002;99(6):1943-1951. (Pubitemid 34525473)
72. Kröger N, Zabelina T, van Biezen A, et al. Donor selection for allogeneic stem cell transplantation in elderly patients with advanced MDS: younger matched-unrelated donor or HLA-identical sibling [abstract]? Blood (ASH Annual Meeting Abstracts). 2010;116: Abstract 912.
73. Bishop MR, Alyea EP 3rd, Cairo MS, et al. National Cancer Institute's First International Workshop on the Biology, Prevention, and Treatment of Relapse after Allogeneic Hematopoietic Stem Cell Transplantation: summary and recommendations from the organizing committee. Biol Blood Marrow Transplant. 2011;17(4):443-454.
74. Campregher PV, Gooley T, Scott BL, et al. Results of donor lymphocyte infusions for relapsed myelodysplastic syndrome after hematopoietic cell transplantation. Bone Marrow Transplant. 2007;40(10):965-971. (Pubitemid 350048195)
75. Depil S, Deconinck E, Milpied N, et al. Donor lymphocyte infusion to treat relapse after allogeneic bone marrow transplantation for myelodysplastic syndrome. Bone Marrow Transplant. 2004;33(5):531-534. (Pubitemid 38425784)
76. Kolb HJ, Schmid C, Tischer J, et al. Allogeneic stem cell transplantation for MDS and sAML following reduced-intensity conditioning and preemptive donor lymphocyte transfusion [abstract]. Blood (ASH Annual Meeting Abstracts). 2006;108: Abstract 324.
77. Schroeder T, Czibere A, Kröger N, et al. Phase II study of azacitidine (Vidaza, Aza) and donor lymphocyte infusions (DLI) as first salvage therapy in patients with acute myeloid leukemia (AML) or myelodysplastic syndromes (MDS) relapsing after allogeneic hematopoietic stem cell transplantation (allo-SCT): final results from AZARELA-trial (NCT-00795548) [abstract]. Blood (ASH Annual Meeting Abstracts). 2011: Abstract 656.
78. Jabbour E, Giralt S, Kantarjian H, et al. Low-dose azacitidine after allogeneic stem cell transplantation for acute leukemia. Cancer. 2009;115(9):1899-1905.
79. de Lima M, Giralt S, Thall PF, et al. Maintenance therapy with low-dose azacitidine after allogeneic hematopoietic stem cell transplantation for recurrent acute myelogeneous leukemia or myelodysplastic syndrome: a dose and schedule finding study. Cancer. 2010;116(23):5420-5431.
80. Atanackovich D, Luetkens T, Kloth B, et al. Cancer-testis antigen expression and its epigenetic modulation in acute myeloid leukemia. Am J Hematol. 2011;86(11):918-922.
81. Goodyear O, Agathanggelou A, Novitzky-Basso I, et al. Induction of a CD8- T-cell response to MAGE cancer testis antigen by combined treatment with azacitidine and sodium valproate in patients with acute myeloid leukemia and myelodysplasia. Blood. 2010;116(11):1908-1918.
82. Stübig T, Badbaran A, Wolschke C, et al. Immune- modulation of CD4+ and CD8+ cells by 5-azacitidine [abstract]. Haematologica. 2010;95(suppl 2):308. Abstract 0738.
83. Kröger N, Bacher U, Bader P, et al. NCI first international workshop on the biology, prevention, and treatment of relapse after allogeneic hematopoietic stem cell transplantation: report from the committee on disease-specific methods and strategies for monitoring relapse following allogeneic stem cell transplantation: II. Chronic leukemias, myeloproliferative neoplasms, and lymphoid malignancies. Biol Blood Marrow Transplant. 2010;16(10):1325-1346.
84. Platzbecker U, Wermke M, Radke J, et al. Azacitidine for treatment of imminent relapse in MDS or AML patients after allogeneic HSCT: results of the RELAZA trial. Leukemia. 2012;26(3):381-389.
85. Thol F, Friesen I, Damm F, et al. Prognostic significance of ASXL1 mutations with myelodysplastic syndromes. J Clin Oncol. 2011;29(18):2499-2506.
86. Center for International Blood & Marrow Transplant Research. Report on state of the art in blood and marrow transplantation: 2009; Slide 9. http://www.cibmtr.org/ReferenceCenter/SlidesReports/ SummarySlides/index.html. Accessed 2009.