{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,5]],"date-time":"2026-02-05T13:37:29Z","timestamp":1770298649141,"version":"3.49.0"},"reference-count":48,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2025,2,28]],"date-time":"2025-02-28T00:00:00Z","timestamp":1740700800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Coordena\u00e7\u00e3o de Aperfei\u00e7oamento de Pessoal de N\u00edvel Superior (CAPES)","award":["88882.332475\/2019-01"],"award-info":[{"award-number":["88882.332475\/2019-01"]}]},{"name":"Funda\u00e7\u00e3o Carlos Chagas Filho de Amparo \u00e0 Pesquisa do Estado do Rio de Janeiro (FAPERJ)","award":["88882.332475\/2019-01"],"award-info":[{"award-number":["88882.332475\/2019-01"]}]},{"name":"Conselho Nacional de Desenvolvimento Cient\u00edfico e Tecnol\u00f3gico (CNPq)","award":["88882.332475\/2019-01"],"award-info":[{"award-number":["88882.332475\/2019-01"]}]},{"name":"Instituto Oswaldo Cruz\/Funda\u00e7\u00e3o Oswaldo Cruz (IOC\/FIOCRUZ)","award":["88882.332475\/2019-01"],"award-info":[{"award-number":["88882.332475\/2019-01"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Nutrients"],"abstract":"<jats:p>Background\/Objectives: Malnutrition and visceral leishmaniasis are major public health problems that are responsible for millions of deaths across many countries. Leishmaniasis development and progression are associated with the host immune status. In this context, malnutrition can directly affect the course of leishmaniasis, impairing several components of the immune system. Moreover, malnutrition directly interferes with the tropism of Leishmania in organs, affecting host susceptibility. Therefore, this work aimed to evaluate the influence of nutritional status on the establishment, progression, and treatment of Leishmania infantum infection in malnourished and refed mice. Methods: BALB\/c mice were fed either a control or restricted diet, infected with L. infantum promastigotes, and treated with meglumine antimoniate, the standard drug for treating visceral leishmaniasis. The effects of infection were evaluated through limiting dilution analysis (LDA). Results: Compared with control mice, malnourished and refed mice presented a lower parasitic load in the spleen, which correlated with spleen atrophy, and the refeeding process partially reversed but did not fully rescue the infection status. Both groups presented a high parasitic load in the liver. Marasmic malnutrition appeared to impair the efficacy of leishmaniasis treatment; however, the refed groups exhibited a robust decrease in the parasite load, which was comparable to that in the control group subjected to treatment. Conclusions: Our data suggested that marasmic malnutrition affects the establishment and progression of Leishmania infection, in addition to reducing the efficacy of standard treatment. Furthermore, the refeeding intervention used did not fully reverse the observed effects. These findings highlight the potential importance of nutritional interventions in the clinical management of visceral leishmaniasis in malnourished populations.<\/jats:p>","DOI":"10.3390\/nu17050849","type":"journal-article","created":{"date-parts":[[2025,2,28]],"date-time":"2025-02-28T10:46:46Z","timestamp":1740739606000},"page":"849","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Impact of Marasmic Malnutrition on Visceral Leishmaniasis: Progression and Treatment Efficacy in a Murine Model"],"prefix":"10.3390","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8953-3537","authenticated-orcid":false,"given":"Taiana","family":"Ferreira-Paes","sequence":"first","affiliation":[{"name":"Laborat\u00f3rio de Bioqu\u00edmica de Tripanosomat\u00eddeos, Instituto Oswaldo Cruz, Funda\u00e7\u00e3o Oswaldo Cruz, Rio de Janeiro 21040-360, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luiza F. O.","family":"Gervazoni","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Bioqu\u00edmica de Tripanosomat\u00eddeos, Instituto Oswaldo Cruz, Funda\u00e7\u00e3o Oswaldo Cruz, Rio de Janeiro 21040-360, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Paula","family":"Seixas-Costa","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Bioqu\u00edmica de Tripanosomat\u00eddeos, Instituto Oswaldo Cruz, Funda\u00e7\u00e3o Oswaldo Cruz, Rio de Janeiro 21040-360, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2017-8602","authenticated-orcid":false,"given":"Paula Mello","family":"De Luca","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Imunoparasitologia Cl\u00ednica, Instituto Oswaldo Cruz, Funda\u00e7\u00e3o Oswaldo Cruz, Rio de Janeiro 21040-360, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1363-3176","authenticated-orcid":false,"given":"Elmo Eduardo","family":"Almeida-Amaral","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Bioqu\u00edmica de Tripanosomat\u00eddeos, Instituto Oswaldo Cruz, Funda\u00e7\u00e3o Oswaldo Cruz, Rio de Janeiro 21040-360, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,2,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Costa, C.H.N., Chang, K.-P., Costa, D.L., and Cunha, F.V.M. (2023). From Infection to Death: An Overview of the Pathogenesis of Visceral Leishmaniasis. Pathogens, 12.","DOI":"10.3390\/pathogens12070969"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Savino, W., Dur\u00e3es, J., Maldonado-Galdeano, C., Perdigon, G., Mendes-da-Cruz, D.A., and Cuervo, P. (2022). Thymus, Undernutrition, and Infection: Approaching Cellular and Molecular Interactions. Front. Nutr., 9.","DOI":"10.3389\/fnut.2022.948488"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1582","DOI":"10.1086\/587658","article-title":"The Interaction between Nutrition and Infection","volume":"46","author":"Katona","year":"2008","journal-title":"Clin. Infect. Dis."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"374","DOI":"10.1590\/S1678-91992009000300003","article-title":"Impact of Malnutrition on Immunity and Infection","volume":"15","author":"Ishikawa","year":"2009","journal-title":"J. Venom. Anim. Toxins Incl. Trop. Dis."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"587","DOI":"10.1111\/j.1365-3024.2009.01117.x","article-title":"Protein-Energy Malnutrition as a Risk Factor for Visceral Leishmaniasis: A Review","volume":"31","author":"Malafaia","year":"2009","journal-title":"Parasite Immunol."},{"key":"ref_6","unstructured":"World Health Organization (2024, October 16). Malnutrition. Available online: https:\/\/www.who.int\/health-topics\/malnutrition."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Imran, M., and Imran, A. (2020). Global Prevalence of Malnutrition: Evidence from Literature. Malnutrition, IntechOpen.","DOI":"10.5772\/intechopen.77861"},{"key":"ref_8","unstructured":"Food and Agriculture Organization of the United Nations (FAO), the International Fund for Agricultural Development (IFAD), the United Nations Children\u2019s Fund (UNICEF), the World Food Programme (WFP), and the World Health Organization (WHO) (2022). The State of Food Security and Nutrition in the World 2022, FAO."},{"key":"ref_9","unstructured":"World Health Organization (2024, October 16). Leishmaniasis. Available online: https:\/\/www.who.int\/news-room\/fact-sheets\/detail\/leishmaniasis."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"481","DOI":"10.1017\/S0031182017002116","article-title":"Chemotherapeutics of Visceral Leishmaniasis: Present and Future Developments","volume":"145","author":"Sundar","year":"2018","journal-title":"Parasitology"},{"key":"ref_11","unstructured":"(2023). Leishmaniasis: Epidemiological Report for the Americas. No.12 (December 2023), PAHO. Available online: https:\/\/iris.paho.org\/handle\/10665.2\/59155."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1186\/s13071-023-06018-2","article-title":"Upregulation of PD-1\/PD-L1 and Downregulation of Immune Signaling Pathways Lead to More Severe Visceral Leishmaniasis in Undernutrition Mice","volume":"17","author":"He","year":"2024","journal-title":"Parasites Vectors"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"591","DOI":"10.4269\/ajtmh.2008.79.591","article-title":"Association of Nutritional Status with the Response to Infection with Leishmania Chagasi","volume":"79","author":"Maciel","year":"2008","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1016\/j.actatropica.2013.06.005","article-title":"Influence of the Nutritional Status in the Clinical and Therapeutical Evolution in Adults and Elderly with American Tegumentary Leishmaniasis","volume":"128","author":"Oliveira","year":"2013","journal-title":"Acta Trop."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Akuffo, H., Costa, C., van Griensven, J., Burza, S., Moreno, J., and Herrero, M. (2018). New Insights into Leishmaniasis in the Immunosuppressed. PLoS Negl. Trop. Dis., 12.","DOI":"10.1371\/journal.pntd.0006375"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1030","DOI":"10.1093\/infdis\/156.6.1030","article-title":"Malnutrition as a Risk Factor for Severe Visceral Leishmaniasis","volume":"156","author":"Cerf","year":"1987","journal-title":"J. Infect. Dis."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Gait\u00e1n-Albarrac\u00edn, F., Losada-Barrag\u00e1n, M., Pinho, N., Azevedo, R., Dur\u00e3es, J., Arcila-Barrera, J.S., Menezes, R.C., Morgado, F.N., Carvalho, V.d.F., and Uma\u00f1a-P\u00e9rez, A. (2021). Malnutrition Aggravates Alterations Observed in the Gut Structure and Immune Response of Mice Infected with Leishmania Infantum. Microorganisms, 9.","DOI":"10.3390\/microorganisms9061270"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"110712","DOI":"10.1016\/j.nut.2019.110712","article-title":"Nutrition, Malnutrition, and Leishmaniasis","volume":"73","author":"Nweze","year":"2020","journal-title":"Nutrition"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"679","DOI":"10.1111\/tmi.12877","article-title":"Causes and Consequences of Higher Leishmania Infantum Burden in Patients with Kala-Azar: A Study of 625 Patients","volume":"22","author":"Zacarias","year":"2017","journal-title":"Trop. Med. Int. Health"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"543","DOI":"10.4269\/ajtmh.2011.10-0321","article-title":"Who Is a Typical Patient with Visceral Leishmaniasis? Characterizing the Demographic and Nutritional Profile of Patients in Brazil, East Africa, and South Asia","volume":"84","author":"Harhay","year":"2011","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"4709","DOI":"10.1128\/IAI.69.8.4709-4718.2001","article-title":"Malnutrition Alters the Innate Immune Response and Increases Early Visceralization Following Leishmania Donovani Infection","volume":"69","author":"Anstead","year":"2001","journal-title":"Infect. Immun."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Ibrahim, M.K., Barnes, J.L., Anstead, G.M., Jimenez, F., Travi, B.L., Peniche, A.G., Osorio, E.Y., Ahuja, S.S., and Melby, P.C. (2013). The Malnutrition-Related Increase in Early Visceralization of Leishmania Donovani Is Associated with a Reduced Number of Lymph Node Phagocytes and Altered Conduit System Flow. PLoS Negl. Trop. Dis., 7.","DOI":"10.1371\/journal.pntd.0002329"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"3098","DOI":"10.1128\/IAI.01778-14","article-title":"Deficiency of Lymph Node-Resident Dendritic Cells (DCs) and Dysregulation of DC Chemoattractants in a Malnourished Mouse Model of Leishmania Donovani Infection","volume":"82","author":"Ibrahim","year":"2014","journal-title":"Infect. Immun."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"28298","DOI":"10.3402\/gha.v8.28298","article-title":"Tackling Malnutrition: A Systematic Review of 15-Year Research Evidence from INDEPTH Health and Demographic Surveillance Systems","volume":"8","author":"Arthur","year":"2015","journal-title":"Glob. Health Action"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Fan, Y., Yao, Q., Liu, Y., Jia, T., Zhang, J., and Jiang, E. (2022). Underlying Causes and Co-Existence of Malnutrition and Infections: An Exceedingly Common Death Risk in Cancer. Front. Nutr., 9.","DOI":"10.3389\/fnut.2022.814095"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"811","DOI":"10.1590\/S0074-02762010000600014","article-title":"Immune Response to Leishmania (Leishmania) Chagasi Infection Is Reduced in Malnourished BALB\/c Mice","volume":"105","author":"Serafim","year":"2010","journal-title":"Mem. Inst. Oswaldo Cruz"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"449","DOI":"10.1016\/j.nut.2013.10.003","article-title":"Leishmania Donovani: Dynamics of L. donovani Evasion of Innate Immune Cell Attack Due to Malnutrition in Visceral Leishmaniasis","volume":"30","author":"Kumar","year":"2014","journal-title":"Nutrition"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Carrillo, E., Jimenez, M.A., Sanchez, C., Cunha, J., Martins, C.M., da Paix\u00e3o Sev\u00e1, A., and Moreno, J. (2014). Protein Malnutrition Impairs the Immune Response and Influences the Severity of Infection in a Hamster Model of Chronic Visceral Leishmaniasis. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0089412"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Losada-Barrag\u00e1n, M., Uma\u00f1a-P\u00e9rez, A., Cuervo-Escobar, S., Berbert, L.R., Porrozzi, R., Morgado, F.N., Mendes-da-Cruz, D.A., Savino, W., S\u00e1nchez-G\u00f3mez, M., and Cuervo, P. (2017). Protein Malnutrition Promotes Dysregulation of Molecules Involved in T Cell Migration in the Thymus of Mice Infected with Leishmania Infantum. Sci. Rep., 7.","DOI":"10.1038\/srep46809"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Ferreira-Paes, T., Seixas-Costa, P., and Almeida-Amaral, E.E. (2021). Validation of a Feed Protocol in a Mouse Model That Mimics Marasmic Malnutrition. Front. Vet. Sci., 8.","DOI":"10.3389\/fvets.2021.757136"},{"key":"ref_31","unstructured":"Shils, M.E. (1999). Modern Nutrition in Health and Disease, Williams & Wilkins. [9th ed.]."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1590\/S1678-91992003000200003","article-title":"Interaction between Infection, Nutrition and Immunity in Tropical Medicine","volume":"9","author":"Pereira","year":"2003","journal-title":"J. Venom. Anim. Toxins Incl. Trop. Dis."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Foolchand, A., Ghazi, T., and Chuturgoon, A.A. (2022). Malnutrition and Dietary Habits Alter the Immune System Which May Consequently Influence SARS-CoV-2 Virulence: A Review. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23052654"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"eaaw6284","DOI":"10.1126\/sciadv.aaw6284","article-title":"Congenital Zika Syndrome Is Associated with Maternal Protein Malnutrition","volume":"6","author":"Pezzuto","year":"2020","journal-title":"Sci. Adv."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Bartelt, L.A., Bolick, D.T., Mayneris-Perxachs, J., Kolling, G.L., Medlock, G.L., Zaenker, E.I., Donowitz, J., Thomas-Beckett, R.V., Rogala, A., and Carroll, I.M. (2017). Cross-Modulation of Pathogen-Specific Pathways Enhances Malnutrition during Enteric Co-Infection with Giardia Lamblia and Enteroaggregative Escherichia Coli. PLoS Pathog., 13.","DOI":"10.1371\/journal.ppat.1006471"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Burgess, S.L., Oka, A., Liu, B., Bolick, D.T., Oakland, D.N., Guerrant, R.L., and Bartelt, L. (2019). Intestinal Parasitic Infection Alters Bone Marrow Derived Dendritic Cell Inflammatory Cytokine Production in Response to Bacterial Endotoxin in a Diet-Dependent Manner. PLoS Negl. Trop. Dis., 13.","DOI":"10.1371\/journal.pntd.0007515"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Losada-Barrag\u00e1n, M., Uma\u00f1a-P\u00e9rez, A., Dur\u00e3es, J., Cuervo-Escobar, S., Rodr\u00edguez-Vega, A., Ribeiro-Gomes, F.L., Berbert, L.R., Morgado, F., Porrozzi, R., and Mendes-da-Cruz, D.A. (2019). Thymic Microenvironment Is Modified by Malnutrition and Leishmania Infantum Infection. Front. Cell Infect. Microbiol., 9.","DOI":"10.3389\/fcimb.2019.00252"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"440","DOI":"10.1186\/s12936-015-0973-z","article-title":"Malaria, Anaemia and under-Nutrition: Three Frequently Co-Existing Conditions among Preschool Children in Rural Rwanda","volume":"14","author":"Kateera","year":"2015","journal-title":"Malar. J."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"120829","DOI":"10.1016\/j.lfs.2022.120829","article-title":"Virulence Factors of Leishmania Parasite: Their Paramount Importance in Unraveling Novel Vaccine Candidates and Therapeutic Targets","volume":"306","author":"Kumari","year":"2022","journal-title":"Life Sci."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"108400","DOI":"10.1016\/j.intimp.2021.108400","article-title":"Exploring the Paradox of Defense between Host and Leishmania Parasite","volume":"102","author":"Kumari","year":"2022","journal-title":"Int. Immunopharmacol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"31","DOI":"10.4103\/0022-3859.930","article-title":"An Overview of Paediatric Leishmaniasis","volume":"49","author":"Kafetzis","year":"2003","journal-title":"J. Postgrad. Med."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Cuervo-Escobar, S., Losada-Barrag\u00e1n, M., Uma\u00f1a-P\u00e9rez, A., Porrozzi, R., Saboia-Vahia, L., Miranda, L.H.M., Morgado, F.N., Menezes, R.C., S\u00e1nchez-G\u00f3mez, M., and Cuervo, P. (2014). T-Cell Populations and Cytokine Expression Are Impaired in Thymus and Spleen of Protein Malnourished BALB\/c Mice Infected with Leishmania Infantum. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0114584"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"2125","DOI":"10.1007\/s10753-014-9947-5","article-title":"Protein Malnutrition Alters Spleen Cell Proliferation and IL-2 and IL-10 Production by Affecting the STAT-1 and STAT-3 Balance","volume":"37","author":"Mello","year":"2014","journal-title":"Inflammation"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1198","DOI":"10.1016\/j.jhep.2016.05.046","article-title":"Malnutrition-Associated Liver Steatosis and ATP Depletion Is Caused by Peroxisomal and Mitochondrial Dysfunction","volume":"65","author":"Ciapaite","year":"2016","journal-title":"J. Hepatol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"104702","DOI":"10.1016\/j.micpath.2020.104702","article-title":"Difference between Kwashiorkor and Marasmus: Comparative Meta-Analysis of Pathogenic Characteristics and Implications for Treatment","volume":"150","author":"Pham","year":"2021","journal-title":"Microb. Pathog."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1348","DOI":"10.1136\/adc.67.11.1348","article-title":"Ultrasonographic Assessment of the Extent of Hepatic Steatosis in Severe Malnutrition","volume":"67","author":"Doherty","year":"1992","journal-title":"Arch. Dis. Child."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"435","DOI":"10.1016\/S0306-3623(97)00268-1","article-title":"The Pharmacology of Leishmaniasis","volume":"30","author":"Reguera","year":"1998","journal-title":"Gen. Pharmacol."},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Scarpini, S., Dondi, A., Totaro, C., Biagi, C., Melchionda, F., Zama, D., Pierantoni, L., Gennari, M., Campagna, C., and Prete, A. (2022). Visceral Leishmaniasis: Epidemiology, Diagnosis, and Treatment Regimens in Different Geographical Areas with a Focus on Pediatrics. Microorganisms, 10.","DOI":"10.3390\/microorganisms10101887"}],"container-title":["Nutrients"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6643\/17\/5\/849\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T16:44:40Z","timestamp":1760028280000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6643\/17\/5\/849"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,2,28]]},"references-count":48,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2025,3]]}},"alternative-id":["nu17050849"],"URL":"https:\/\/doi.org\/10.3390\/nu17050849","relation":{},"ISSN":["2072-6643"],"issn-type":[{"value":"2072-6643","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,2,28]]}}}