Exploring the interplay of complex carbohydrate intake, the microbiome CAZymes pool and short-chain fatty acid production in the human gut: Insights from different cohorts in the A...
- Autores
- Trotta, Milagros; Agustini, Camila; Rohr, Cristian Oscar; Ame, Ricardo Martín; Giordano, Laureano Jose; Balderramo, Domingo; Romagnoli, Pablo Alberto; Vazquez, Martin
- Año de publicación
- 2025
- Idioma
- inglés
- Tipo de recurso
- artículo
- Estado
- versión publicada
- Descripción
- Carbohydrate-active enzyme (CAZyme) activity in the gut microbiome has significant implications for health, including the release of nutrients otherwise inaccessible to the host and the enhancement of digestive efficiency. The primary end products of indigestible carbohydrate fermentation are short-chain fatty acids (SCFAs). We hypothesized that increased dietary fiber consumption could lead to greater SCFA production in the human gut. To investigate this, we examined the relationship between complex carbohydrate intake, CAZyme activity in the human gut microbiome, and SCFA production at the whole metagenomic level across three cohorts: a healthy reference-controlled cohort, an average cohort of individuals living in industrialized cities, and a cohort of patients with inflammatory bowel disease (IBD). Metagenomic sequencing and bioinformatic analyses were utilized to assess the diversity, abundance, and functionality of CAZymes, as well as the metabolic capacity for SCFA production. The average cohort exhibited higher alpha diversity of CAZyme families compared to the reference-controlled cohort, although subfamily composition was similar between both. A moderate negative correlation was identified between CAZyme abundance and SCFA production, indicating that a higher number of these enzymes does not directly translate to increased SCFA synthesis. In IBD patients, a decrease in the diversity and composition of CAZyme subfamilies was observed, suggesting a disruption in enzymatic functions associated with the disease. However, the overall functionality of CAZymes remained relatively stable across different health conditions, highlighting the resilience of the gut microbiome for these functions. These findings deepen our understanding of the gut microbiome’s role in health and disease, emphasizing that despite variations in microbial diversity, key enzymatic functions persist. The study underscores the complexity of the non-linear relationship between complex carbohydrate metabolism and SCFA production, laying the groundwork for future research on microbiome-targeted therapeutic/dietary profile interventions in both non-disease and chronic diseases conditions.
Fil: Trotta, Milagros. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Heritas S.a (heritas S.a);
Fil: Agustini, Camila. Heritas S.a (heritas S.a);
Fil: Rohr, Cristian Oscar. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Heritas S.a (heritas S.a);
Fil: Ame, Ricardo Martín. Hospital Privado Universitario de Córdoba; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Grupo Vinculado Centro de Investigación en Medicina Traslacional Severo R. Amuchástegui - Cimetsa | Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Grupo Vinculado Centro de Investigación en Medicina Traslacional Severo R. Amuchástegui - Cimetsa | Instituto de Investigación Médica Mercedes y Martín Ferreyra. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Grupo Vinculado Centro de Investigación en Medicina Traslacional Severo R. Amuchástegui - Cimetsa; Argentina
Fil: Giordano, Laureano Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Grupo Vinculado Centro de Investigación en Medicina Traslacional Severo R. Amuchástegui - Cimetsa | Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Grupo Vinculado Centro de Investigación en Medicina Traslacional Severo R. Amuchástegui - Cimetsa | Instituto de Investigación Médica Mercedes y Martín Ferreyra. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Grupo Vinculado Centro de Investigación en Medicina Traslacional Severo R. Amuchástegui - Cimetsa; Argentina
Fil: Balderramo, Domingo. Hospital Privado Universitario de Córdoba; Argentina
Fil: Romagnoli, Pablo Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Grupo Vinculado Centro de Investigación en Medicina Traslacional Severo R. Amuchástegui - Cimetsa | Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Grupo Vinculado Centro de Investigación en Medicina Traslacional Severo R. Amuchástegui - Cimetsa | Instituto de Investigación Médica Mercedes y Martín Ferreyra. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Grupo Vinculado Centro de Investigación en Medicina Traslacional Severo R. Amuchástegui - Cimetsa; Argentina
Fil: Vazquez, Martin. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Heritas S.a (heritas S.a); - Materia
-
GUT MICROBIOME
CAZYMES
SCFAS
IBD
MICROBIAL DIVERSITY
ENZYMATIC FUNCTIONALITY
MICROBIOME RESILIENCE - Nivel de accesibilidad
- acceso abierto
- Condiciones de uso
- https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
- Repositorio
.jpg)
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/274101
Ver los metadatos del registro completo
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Exploring the interplay of complex carbohydrate intake, the microbiome CAZymes pool and short-chain fatty acid production in the human gut: Insights from different cohorts in the Argentine populationTrotta, MilagrosAgustini, CamilaRohr, Cristian OscarAme, Ricardo MartínGiordano, Laureano JoseBalderramo, DomingoRomagnoli, Pablo AlbertoVazquez, MartinGUT MICROBIOMECAZYMESSCFASIBDMICROBIAL DIVERSITYENZYMATIC FUNCTIONALITYMICROBIOME RESILIENCEhttps://purl.org/becyt/ford/3.2https://purl.org/becyt/ford/3Carbohydrate-active enzyme (CAZyme) activity in the gut microbiome has significant implications for health, including the release of nutrients otherwise inaccessible to the host and the enhancement of digestive efficiency. The primary end products of indigestible carbohydrate fermentation are short-chain fatty acids (SCFAs). We hypothesized that increased dietary fiber consumption could lead to greater SCFA production in the human gut. To investigate this, we examined the relationship between complex carbohydrate intake, CAZyme activity in the human gut microbiome, and SCFA production at the whole metagenomic level across three cohorts: a healthy reference-controlled cohort, an average cohort of individuals living in industrialized cities, and a cohort of patients with inflammatory bowel disease (IBD). Metagenomic sequencing and bioinformatic analyses were utilized to assess the diversity, abundance, and functionality of CAZymes, as well as the metabolic capacity for SCFA production. The average cohort exhibited higher alpha diversity of CAZyme families compared to the reference-controlled cohort, although subfamily composition was similar between both. A moderate negative correlation was identified between CAZyme abundance and SCFA production, indicating that a higher number of these enzymes does not directly translate to increased SCFA synthesis. In IBD patients, a decrease in the diversity and composition of CAZyme subfamilies was observed, suggesting a disruption in enzymatic functions associated with the disease. However, the overall functionality of CAZymes remained relatively stable across different health conditions, highlighting the resilience of the gut microbiome for these functions. These findings deepen our understanding of the gut microbiome’s role in health and disease, emphasizing that despite variations in microbial diversity, key enzymatic functions persist. The study underscores the complexity of the non-linear relationship between complex carbohydrate metabolism and SCFA production, laying the groundwork for future research on microbiome-targeted therapeutic/dietary profile interventions in both non-disease and chronic diseases conditions.Fil: Trotta, Milagros. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Heritas S.a (heritas S.a);Fil: Agustini, Camila. Heritas S.a (heritas S.a);Fil: Rohr, Cristian Oscar. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Heritas S.a (heritas S.a);Fil: Ame, Ricardo Martín. Hospital Privado Universitario de Córdoba; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Grupo Vinculado Centro de Investigación en Medicina Traslacional Severo R. Amuchástegui - Cimetsa | Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Grupo Vinculado Centro de Investigación en Medicina Traslacional Severo R. Amuchástegui - Cimetsa | Instituto de Investigación Médica Mercedes y Martín Ferreyra. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Grupo Vinculado Centro de Investigación en Medicina Traslacional Severo R. Amuchástegui - Cimetsa; ArgentinaFil: Giordano, Laureano Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Grupo Vinculado Centro de Investigación en Medicina Traslacional Severo R. Amuchástegui - Cimetsa | Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Grupo Vinculado Centro de Investigación en Medicina Traslacional Severo R. Amuchástegui - Cimetsa | Instituto de Investigación Médica Mercedes y Martín Ferreyra. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Grupo Vinculado Centro de Investigación en Medicina Traslacional Severo R. Amuchástegui - Cimetsa; ArgentinaFil: Balderramo, Domingo. Hospital Privado Universitario de Córdoba; ArgentinaFil: Romagnoli, Pablo Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Grupo Vinculado Centro de Investigación en Medicina Traslacional Severo R. Amuchástegui - Cimetsa | Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Grupo Vinculado Centro de Investigación en Medicina Traslacional Severo R. Amuchástegui - Cimetsa | Instituto de Investigación Médica Mercedes y Martín Ferreyra. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Grupo Vinculado Centro de Investigación en Medicina Traslacional Severo R. Amuchástegui - Cimetsa; ArgentinaFil: Vazquez, Martin. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Heritas S.a (heritas S.a);Cold Spring Harbor Laboratory Press2025-05info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/274101Trotta, Milagros; Agustini, Camila; Rohr, Cristian Oscar; Ame, Ricardo Martín; Giordano, Laureano Jose; et al.; Exploring the interplay of complex carbohydrate intake, the microbiome CAZymes pool and short-chain fatty acid production in the human gut: Insights from different cohorts in the Argentine population; Cold Spring Harbor Laboratory Press; bioXriv; 5-2025; 1-192692-8205CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.biorxiv.org/content/10.1101/2025.05.15.654276v1info:eu-repo/semantics/altIdentifier/doi/10.1101/2025.05.15.654276info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-11-05T09:43:06Zoai:ri.conicet.gov.ar:11336/274101instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-11-05 09:43:06.567CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse |
| dc.title.none.fl_str_mv |
Exploring the interplay of complex carbohydrate intake, the microbiome CAZymes pool and short-chain fatty acid production in the human gut: Insights from different cohorts in the Argentine population |
| title |
Exploring the interplay of complex carbohydrate intake, the microbiome CAZymes pool and short-chain fatty acid production in the human gut: Insights from different cohorts in the Argentine population |
| spellingShingle |
Exploring the interplay of complex carbohydrate intake, the microbiome CAZymes pool and short-chain fatty acid production in the human gut: Insights from different cohorts in the Argentine population Trotta, Milagros GUT MICROBIOME CAZYMES SCFAS IBD MICROBIAL DIVERSITY ENZYMATIC FUNCTIONALITY MICROBIOME RESILIENCE |
| title_short |
Exploring the interplay of complex carbohydrate intake, the microbiome CAZymes pool and short-chain fatty acid production in the human gut: Insights from different cohorts in the Argentine population |
| title_full |
Exploring the interplay of complex carbohydrate intake, the microbiome CAZymes pool and short-chain fatty acid production in the human gut: Insights from different cohorts in the Argentine population |
| title_fullStr |
Exploring the interplay of complex carbohydrate intake, the microbiome CAZymes pool and short-chain fatty acid production in the human gut: Insights from different cohorts in the Argentine population |
| title_full_unstemmed |
Exploring the interplay of complex carbohydrate intake, the microbiome CAZymes pool and short-chain fatty acid production in the human gut: Insights from different cohorts in the Argentine population |
| title_sort |
Exploring the interplay of complex carbohydrate intake, the microbiome CAZymes pool and short-chain fatty acid production in the human gut: Insights from different cohorts in the Argentine population |
| dc.creator.none.fl_str_mv |
Trotta, Milagros Agustini, Camila Rohr, Cristian Oscar Ame, Ricardo Martín Giordano, Laureano Jose Balderramo, Domingo Romagnoli, Pablo Alberto Vazquez, Martin |
| author |
Trotta, Milagros |
| author_facet |
Trotta, Milagros Agustini, Camila Rohr, Cristian Oscar Ame, Ricardo Martín Giordano, Laureano Jose Balderramo, Domingo Romagnoli, Pablo Alberto Vazquez, Martin |
| author_role |
author |
| author2 |
Agustini, Camila Rohr, Cristian Oscar Ame, Ricardo Martín Giordano, Laureano Jose Balderramo, Domingo Romagnoli, Pablo Alberto Vazquez, Martin |
| author2_role |
author author author author author author author |
| dc.subject.none.fl_str_mv |
GUT MICROBIOME CAZYMES SCFAS IBD MICROBIAL DIVERSITY ENZYMATIC FUNCTIONALITY MICROBIOME RESILIENCE |
| topic |
GUT MICROBIOME CAZYMES SCFAS IBD MICROBIAL DIVERSITY ENZYMATIC FUNCTIONALITY MICROBIOME RESILIENCE |
| purl_subject.fl_str_mv |
https://purl.org/becyt/ford/3.2 https://purl.org/becyt/ford/3 |
| dc.description.none.fl_txt_mv |
Carbohydrate-active enzyme (CAZyme) activity in the gut microbiome has significant implications for health, including the release of nutrients otherwise inaccessible to the host and the enhancement of digestive efficiency. The primary end products of indigestible carbohydrate fermentation are short-chain fatty acids (SCFAs). We hypothesized that increased dietary fiber consumption could lead to greater SCFA production in the human gut. To investigate this, we examined the relationship between complex carbohydrate intake, CAZyme activity in the human gut microbiome, and SCFA production at the whole metagenomic level across three cohorts: a healthy reference-controlled cohort, an average cohort of individuals living in industrialized cities, and a cohort of patients with inflammatory bowel disease (IBD). Metagenomic sequencing and bioinformatic analyses were utilized to assess the diversity, abundance, and functionality of CAZymes, as well as the metabolic capacity for SCFA production. The average cohort exhibited higher alpha diversity of CAZyme families compared to the reference-controlled cohort, although subfamily composition was similar between both. A moderate negative correlation was identified between CAZyme abundance and SCFA production, indicating that a higher number of these enzymes does not directly translate to increased SCFA synthesis. In IBD patients, a decrease in the diversity and composition of CAZyme subfamilies was observed, suggesting a disruption in enzymatic functions associated with the disease. However, the overall functionality of CAZymes remained relatively stable across different health conditions, highlighting the resilience of the gut microbiome for these functions. These findings deepen our understanding of the gut microbiome’s role in health and disease, emphasizing that despite variations in microbial diversity, key enzymatic functions persist. The study underscores the complexity of the non-linear relationship between complex carbohydrate metabolism and SCFA production, laying the groundwork for future research on microbiome-targeted therapeutic/dietary profile interventions in both non-disease and chronic diseases conditions. Fil: Trotta, Milagros. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Heritas S.a (heritas S.a); Fil: Agustini, Camila. Heritas S.a (heritas S.a); Fil: Rohr, Cristian Oscar. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Heritas S.a (heritas S.a); Fil: Ame, Ricardo Martín. Hospital Privado Universitario de Córdoba; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Grupo Vinculado Centro de Investigación en Medicina Traslacional Severo R. Amuchástegui - Cimetsa | Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Grupo Vinculado Centro de Investigación en Medicina Traslacional Severo R. Amuchástegui - Cimetsa | Instituto de Investigación Médica Mercedes y Martín Ferreyra. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Grupo Vinculado Centro de Investigación en Medicina Traslacional Severo R. Amuchástegui - Cimetsa; Argentina Fil: Giordano, Laureano Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Grupo Vinculado Centro de Investigación en Medicina Traslacional Severo R. Amuchástegui - Cimetsa | Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Grupo Vinculado Centro de Investigación en Medicina Traslacional Severo R. Amuchástegui - Cimetsa | Instituto de Investigación Médica Mercedes y Martín Ferreyra. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Grupo Vinculado Centro de Investigación en Medicina Traslacional Severo R. Amuchástegui - Cimetsa; Argentina Fil: Balderramo, Domingo. Hospital Privado Universitario de Córdoba; Argentina Fil: Romagnoli, Pablo Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Grupo Vinculado Centro de Investigación en Medicina Traslacional Severo R. Amuchástegui - Cimetsa | Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Grupo Vinculado Centro de Investigación en Medicina Traslacional Severo R. Amuchástegui - Cimetsa | Instituto de Investigación Médica Mercedes y Martín Ferreyra. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Grupo Vinculado Centro de Investigación en Medicina Traslacional Severo R. Amuchástegui - Cimetsa; Argentina Fil: Vazquez, Martin. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Heritas S.a (heritas S.a); |
| description |
Carbohydrate-active enzyme (CAZyme) activity in the gut microbiome has significant implications for health, including the release of nutrients otherwise inaccessible to the host and the enhancement of digestive efficiency. The primary end products of indigestible carbohydrate fermentation are short-chain fatty acids (SCFAs). We hypothesized that increased dietary fiber consumption could lead to greater SCFA production in the human gut. To investigate this, we examined the relationship between complex carbohydrate intake, CAZyme activity in the human gut microbiome, and SCFA production at the whole metagenomic level across three cohorts: a healthy reference-controlled cohort, an average cohort of individuals living in industrialized cities, and a cohort of patients with inflammatory bowel disease (IBD). Metagenomic sequencing and bioinformatic analyses were utilized to assess the diversity, abundance, and functionality of CAZymes, as well as the metabolic capacity for SCFA production. The average cohort exhibited higher alpha diversity of CAZyme families compared to the reference-controlled cohort, although subfamily composition was similar between both. A moderate negative correlation was identified between CAZyme abundance and SCFA production, indicating that a higher number of these enzymes does not directly translate to increased SCFA synthesis. In IBD patients, a decrease in the diversity and composition of CAZyme subfamilies was observed, suggesting a disruption in enzymatic functions associated with the disease. However, the overall functionality of CAZymes remained relatively stable across different health conditions, highlighting the resilience of the gut microbiome for these functions. These findings deepen our understanding of the gut microbiome’s role in health and disease, emphasizing that despite variations in microbial diversity, key enzymatic functions persist. The study underscores the complexity of the non-linear relationship between complex carbohydrate metabolism and SCFA production, laying the groundwork for future research on microbiome-targeted therapeutic/dietary profile interventions in both non-disease and chronic diseases conditions. |
| publishDate |
2025 |
| dc.date.none.fl_str_mv |
2025-05 |
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info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://purl.org/coar/resource_type/c_6501 info:ar-repo/semantics/articulo |
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article |
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publishedVersion |
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http://hdl.handle.net/11336/274101 Trotta, Milagros; Agustini, Camila; Rohr, Cristian Oscar; Ame, Ricardo Martín; Giordano, Laureano Jose; et al.; Exploring the interplay of complex carbohydrate intake, the microbiome CAZymes pool and short-chain fatty acid production in the human gut: Insights from different cohorts in the Argentine population; Cold Spring Harbor Laboratory Press; bioXriv; 5-2025; 1-19 2692-8205 CONICET Digital CONICET |
| url |
http://hdl.handle.net/11336/274101 |
| identifier_str_mv |
Trotta, Milagros; Agustini, Camila; Rohr, Cristian Oscar; Ame, Ricardo Martín; Giordano, Laureano Jose; et al.; Exploring the interplay of complex carbohydrate intake, the microbiome CAZymes pool and short-chain fatty acid production in the human gut: Insights from different cohorts in the Argentine population; Cold Spring Harbor Laboratory Press; bioXriv; 5-2025; 1-19 2692-8205 CONICET Digital CONICET |
| dc.language.none.fl_str_mv |
eng |
| language |
eng |
| dc.relation.none.fl_str_mv |
info:eu-repo/semantics/altIdentifier/url/https://www.biorxiv.org/content/10.1101/2025.05.15.654276v1 info:eu-repo/semantics/altIdentifier/doi/10.1101/2025.05.15.654276 |
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info:eu-repo/semantics/openAccess https://creativecommons.org/licenses/by-nc-nd/2.5/ar/ |
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openAccess |
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https://creativecommons.org/licenses/by-nc-nd/2.5/ar/ |
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application/pdf application/pdf |
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Cold Spring Harbor Laboratory Press |
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Cold Spring Harbor Laboratory Press |
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Consejo Nacional de Investigaciones Científicas y Técnicas |
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CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas |
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dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar |
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13.082534 |