N-glycan biosynthesis inhibitors induce in vitro anticancer activity in colorectal cancer cells
During malignant transformation, changes in the expression profile of glycans may be involved in a variety of events, including the loss of cell–cell and cell–matrix adhesion, migration, invasion, and evasion of apoptosis. Therefore, modulation of glycan expression with drugs has promising therapeut...
Gespeichert in:
| Veröffentlicht in: | Journal of cellular biochemistry 2012-09, Vol.113 (9), p.2957-2966 |
|---|---|
| Hauptverfasser: | , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 2966 |
|---|---|
| container_issue | 9 |
| container_start_page | 2957 |
| container_title | Journal of cellular biochemistry |
| container_volume | 113 |
| creator | de-Freitas-Junior, Julio Cesar Madureira Bastos, Lilian Golçalves Freire-Neto, Carlos Alberto Rocher, Bárbara Du Abdelhay, Eliana Saul Furquim Werneck Morgado-Díaz, José Andrés |
| description | During malignant transformation, changes in the expression profile of glycans may be involved in a variety of events, including the loss of cell–cell and cell–matrix adhesion, migration, invasion, and evasion of apoptosis. Therefore, modulation of glycan expression with drugs has promising therapeutic potential for various cancer types. In this study, we investigated the in vitro anticancer activity of the N‐glycan biosynthesis inhibitors (swainsonine and tunicamycin) in cells derived from colorectal cancer (CRC). We also examined whether these inhibitors are able to induce radiosensitization and toxicity when used in combination with cisplatin or irinotecan, two current anticancer drugs. Our results show that treatment with tunicamycin inhibits cellular mechanisms related to the malignant phenotype, such as anchorage‐dependent and anchorage‐independent colony formation, migration and invasion, in undifferentiated HCT‐116 colon cancer cells, whereas swainsonine only inhibits cell migration. We also observed that tunicamycin, but not swainsonine, caused radiosensitivity in HCT‐116 cells. Moreover, the combination of swainsonine with cisplatin or irinotecan enhanced their toxicity in HCT‐116 cells, while the combination of tunicamycin with these drugs had no effect. Given these results, we suggest that the modulation of N‐glycan biosynthesis appears to be a potential therapeutic tool for CRC treatment because inhibition of this process induced anticancer activity in vitro. Additionally, the inhibition of the N‐glycan biosynthesis in combination with chemotherapic drugs is a promising therapeutic strategy for enhancing radiation therapy. J. Cell. Biochem. 113: 2957–2966, 2012. © 2012 Wiley Periodicals, Inc. |
| doi_str_mv | 10.1002/jcb.24173 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_journals_3113516599</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>3113516599</sourcerecordid><originalsourceid>FETCH-LOGICAL-c4573-26c179a62dcca6780cce2278705a99c93c5bd5cc5f14cddd14c3752c351ed3f53</originalsourceid><addsrcrecordid>eNp1kE1PAjEQhhujEUQP_gGziScPC_3YbulRiKKEYEwwHpvubJHisovtou6_twh48zIzmXnmncyL0CXBXYIx7S0h69KECHaE2gRLESdpkhyjNhYMx5QR2kJn3i8xxlIyeopalHJOZSLbSE3jt6IBXUaZrXxT1gvjrY9subCZrSu3LfMNmJCiT1u7KtJlbQMPxkUaahuazXYIVVE5A7Uuov0UTFH4c3Qy14U3F_vcQS_3d7PhQzx5Gj0ObycxJFywmKZAhNQpzQF0KvoYwFAq-gJzLSVIBjzLOQCfkwTyPA-RCU6BcWJyNuesg653umtXfWyMr9Wy2rgynFSMkIClPPzeQTc7ClzlvTNztXZ2pV2jCFZbK1WwUv1aGdirveImW5n8jzx4F4DeDviyhWn-V1Lj4eAgGe82rK_N99-Gdu8qFeEf9Todqel4MOtPn4Wi7AdmpI1S</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>3113516599</pqid></control><display><type>article</type><title>N-glycan biosynthesis inhibitors induce in vitro anticancer activity in colorectal cancer cells</title><source>MEDLINE</source><source>Wiley Journals</source><creator>de-Freitas-Junior, Julio Cesar Madureira ; Bastos, Lilian Golçalves ; Freire-Neto, Carlos Alberto ; Rocher, Bárbara Du ; Abdelhay, Eliana Saul Furquim Werneck ; Morgado-Díaz, José Andrés</creator><creatorcontrib>de-Freitas-Junior, Julio Cesar Madureira ; Bastos, Lilian Golçalves ; Freire-Neto, Carlos Alberto ; Rocher, Bárbara Du ; Abdelhay, Eliana Saul Furquim Werneck ; Morgado-Díaz, José Andrés</creatorcontrib><description>During malignant transformation, changes in the expression profile of glycans may be involved in a variety of events, including the loss of cell–cell and cell–matrix adhesion, migration, invasion, and evasion of apoptosis. Therefore, modulation of glycan expression with drugs has promising therapeutic potential for various cancer types. In this study, we investigated the in vitro anticancer activity of the N‐glycan biosynthesis inhibitors (swainsonine and tunicamycin) in cells derived from colorectal cancer (CRC). We also examined whether these inhibitors are able to induce radiosensitization and toxicity when used in combination with cisplatin or irinotecan, two current anticancer drugs. Our results show that treatment with tunicamycin inhibits cellular mechanisms related to the malignant phenotype, such as anchorage‐dependent and anchorage‐independent colony formation, migration and invasion, in undifferentiated HCT‐116 colon cancer cells, whereas swainsonine only inhibits cell migration. We also observed that tunicamycin, but not swainsonine, caused radiosensitivity in HCT‐116 cells. Moreover, the combination of swainsonine with cisplatin or irinotecan enhanced their toxicity in HCT‐116 cells, while the combination of tunicamycin with these drugs had no effect. Given these results, we suggest that the modulation of N‐glycan biosynthesis appears to be a potential therapeutic tool for CRC treatment because inhibition of this process induced anticancer activity in vitro. Additionally, the inhibition of the N‐glycan biosynthesis in combination with chemotherapic drugs is a promising therapeutic strategy for enhancing radiation therapy. J. Cell. Biochem. 113: 2957–2966, 2012. © 2012 Wiley Periodicals, Inc.</description><identifier>ISSN: 0730-2312</identifier><identifier>EISSN: 1097-4644</identifier><identifier>DOI: 10.1002/jcb.24173</identifier><identifier>PMID: 22552949</identifier><language>eng</language><publisher>Hoboken: Wiley Subscription Services, Inc., A Wiley Company</publisher><subject>Anchorages ; Anticancer properties ; Antineoplastic drugs ; Antitumor activity ; Apoptosis ; Biosynthesis ; Blotting, Western ; Caco-2 Cells ; Camptothecin - analogs & derivatives ; Camptothecin - pharmacology ; Cancer therapies ; Cell migration ; Cell Movement - drug effects ; Cell Survival - drug effects ; Cisplatin ; Cisplatin - pharmacology ; Colon cancer ; COLORECTAL CANCER ; Colorectal carcinoma ; Colorectal Neoplasms - metabolism ; Drugs ; Flow Cytometry ; Glycan ; HCT116 Cells ; HT29 Cells ; Humans ; Inhibitors ; Irinotecan ; Modulation ; N-GLYCOSYLATION ; Phenotypes ; Polysaccharides ; Polysaccharides - metabolism ; Radiation therapy ; RADIOSENSITIVITY ; Radiosensitization ; Swainsonine - pharmacology ; Toxicity ; TUNICAMYCIN ; Tunicamycin - pharmacology</subject><ispartof>Journal of cellular biochemistry, 2012-09, Vol.113 (9), p.2957-2966</ispartof><rights>Copyright © 2012 Wiley Periodicals, Inc.</rights><rights>Copyright Wiley Subscription Services, Inc. Sep 2012</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4573-26c179a62dcca6780cce2278705a99c93c5bd5cc5f14cddd14c3752c351ed3f53</citedby><cites>FETCH-LOGICAL-c4573-26c179a62dcca6780cce2278705a99c93c5bd5cc5f14cddd14c3752c351ed3f53</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Fjcb.24173$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fjcb.24173$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,27903,27904,45553,45554</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/22552949$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>de-Freitas-Junior, Julio Cesar Madureira</creatorcontrib><creatorcontrib>Bastos, Lilian Golçalves</creatorcontrib><creatorcontrib>Freire-Neto, Carlos Alberto</creatorcontrib><creatorcontrib>Rocher, Bárbara Du</creatorcontrib><creatorcontrib>Abdelhay, Eliana Saul Furquim Werneck</creatorcontrib><creatorcontrib>Morgado-Díaz, José Andrés</creatorcontrib><title>N-glycan biosynthesis inhibitors induce in vitro anticancer activity in colorectal cancer cells</title><title>Journal of cellular biochemistry</title><addtitle>J. Cell. Biochem</addtitle><description>During malignant transformation, changes in the expression profile of glycans may be involved in a variety of events, including the loss of cell–cell and cell–matrix adhesion, migration, invasion, and evasion of apoptosis. Therefore, modulation of glycan expression with drugs has promising therapeutic potential for various cancer types. In this study, we investigated the in vitro anticancer activity of the N‐glycan biosynthesis inhibitors (swainsonine and tunicamycin) in cells derived from colorectal cancer (CRC). We also examined whether these inhibitors are able to induce radiosensitization and toxicity when used in combination with cisplatin or irinotecan, two current anticancer drugs. Our results show that treatment with tunicamycin inhibits cellular mechanisms related to the malignant phenotype, such as anchorage‐dependent and anchorage‐independent colony formation, migration and invasion, in undifferentiated HCT‐116 colon cancer cells, whereas swainsonine only inhibits cell migration. We also observed that tunicamycin, but not swainsonine, caused radiosensitivity in HCT‐116 cells. Moreover, the combination of swainsonine with cisplatin or irinotecan enhanced their toxicity in HCT‐116 cells, while the combination of tunicamycin with these drugs had no effect. Given these results, we suggest that the modulation of N‐glycan biosynthesis appears to be a potential therapeutic tool for CRC treatment because inhibition of this process induced anticancer activity in vitro. Additionally, the inhibition of the N‐glycan biosynthesis in combination with chemotherapic drugs is a promising therapeutic strategy for enhancing radiation therapy. J. Cell. Biochem. 113: 2957–2966, 2012. © 2012 Wiley Periodicals, Inc.</description><subject>Anchorages</subject><subject>Anticancer properties</subject><subject>Antineoplastic drugs</subject><subject>Antitumor activity</subject><subject>Apoptosis</subject><subject>Biosynthesis</subject><subject>Blotting, Western</subject><subject>Caco-2 Cells</subject><subject>Camptothecin - analogs & derivatives</subject><subject>Camptothecin - pharmacology</subject><subject>Cancer therapies</subject><subject>Cell migration</subject><subject>Cell Movement - drug effects</subject><subject>Cell Survival - drug effects</subject><subject>Cisplatin</subject><subject>Cisplatin - pharmacology</subject><subject>Colon cancer</subject><subject>COLORECTAL CANCER</subject><subject>Colorectal carcinoma</subject><subject>Colorectal Neoplasms - metabolism</subject><subject>Drugs</subject><subject>Flow Cytometry</subject><subject>Glycan</subject><subject>HCT116 Cells</subject><subject>HT29 Cells</subject><subject>Humans</subject><subject>Inhibitors</subject><subject>Irinotecan</subject><subject>Modulation</subject><subject>N-GLYCOSYLATION</subject><subject>Phenotypes</subject><subject>Polysaccharides</subject><subject>Polysaccharides - metabolism</subject><subject>Radiation therapy</subject><subject>RADIOSENSITIVITY</subject><subject>Radiosensitization</subject><subject>Swainsonine - pharmacology</subject><subject>Toxicity</subject><subject>TUNICAMYCIN</subject><subject>Tunicamycin - pharmacology</subject><issn>0730-2312</issn><issn>1097-4644</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kE1PAjEQhhujEUQP_gGziScPC_3YbulRiKKEYEwwHpvubJHisovtou6_twh48zIzmXnmncyL0CXBXYIx7S0h69KECHaE2gRLESdpkhyjNhYMx5QR2kJn3i8xxlIyeopalHJOZSLbSE3jt6IBXUaZrXxT1gvjrY9subCZrSu3LfMNmJCiT1u7KtJlbQMPxkUaahuazXYIVVE5A7Uuov0UTFH4c3Qy14U3F_vcQS_3d7PhQzx5Gj0ObycxJFywmKZAhNQpzQF0KvoYwFAq-gJzLSVIBjzLOQCfkwTyPA-RCU6BcWJyNuesg653umtXfWyMr9Wy2rgynFSMkIClPPzeQTc7ClzlvTNztXZ2pV2jCFZbK1WwUv1aGdirveImW5n8jzx4F4DeDviyhWn-V1Lj4eAgGe82rK_N99-Gdu8qFeEf9Todqel4MOtPn4Wi7AdmpI1S</recordid><startdate>201209</startdate><enddate>201209</enddate><creator>de-Freitas-Junior, Julio Cesar Madureira</creator><creator>Bastos, Lilian Golçalves</creator><creator>Freire-Neto, Carlos Alberto</creator><creator>Rocher, Bárbara Du</creator><creator>Abdelhay, Eliana Saul Furquim Werneck</creator><creator>Morgado-Díaz, José Andrés</creator><general>Wiley Subscription Services, Inc., A Wiley Company</general><general>Wiley Subscription Services, Inc</general><scope>BSCLL</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7QP</scope><scope>7QR</scope><scope>7T7</scope><scope>7TK</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>K9.</scope><scope>M7N</scope><scope>P64</scope></search><sort><creationdate>201209</creationdate><title>N-glycan biosynthesis inhibitors induce in vitro anticancer activity in colorectal cancer cells</title><author>de-Freitas-Junior, Julio Cesar Madureira ; Bastos, Lilian Golçalves ; Freire-Neto, Carlos Alberto ; Rocher, Bárbara Du ; Abdelhay, Eliana Saul Furquim Werneck ; Morgado-Díaz, José Andrés</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4573-26c179a62dcca6780cce2278705a99c93c5bd5cc5f14cddd14c3752c351ed3f53</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2012</creationdate><topic>Anchorages</topic><topic>Anticancer properties</topic><topic>Antineoplastic drugs</topic><topic>Antitumor activity</topic><topic>Apoptosis</topic><topic>Biosynthesis</topic><topic>Blotting, Western</topic><topic>Caco-2 Cells</topic><topic>Camptothecin - analogs & derivatives</topic><topic>Camptothecin - pharmacology</topic><topic>Cancer therapies</topic><topic>Cell migration</topic><topic>Cell Movement - drug effects</topic><topic>Cell Survival - drug effects</topic><topic>Cisplatin</topic><topic>Cisplatin - pharmacology</topic><topic>Colon cancer</topic><topic>COLORECTAL CANCER</topic><topic>Colorectal carcinoma</topic><topic>Colorectal Neoplasms - metabolism</topic><topic>Drugs</topic><topic>Flow Cytometry</topic><topic>Glycan</topic><topic>HCT116 Cells</topic><topic>HT29 Cells</topic><topic>Humans</topic><topic>Inhibitors</topic><topic>Irinotecan</topic><topic>Modulation</topic><topic>N-GLYCOSYLATION</topic><topic>Phenotypes</topic><topic>Polysaccharides</topic><topic>Polysaccharides - metabolism</topic><topic>Radiation therapy</topic><topic>RADIOSENSITIVITY</topic><topic>Radiosensitization</topic><topic>Swainsonine - pharmacology</topic><topic>Toxicity</topic><topic>TUNICAMYCIN</topic><topic>Tunicamycin - pharmacology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>de-Freitas-Junior, Julio Cesar Madureira</creatorcontrib><creatorcontrib>Bastos, Lilian Golçalves</creatorcontrib><creatorcontrib>Freire-Neto, Carlos Alberto</creatorcontrib><creatorcontrib>Rocher, Bárbara Du</creatorcontrib><creatorcontrib>Abdelhay, Eliana Saul Furquim Werneck</creatorcontrib><creatorcontrib>Morgado-Díaz, José Andrés</creatorcontrib><collection>Istex</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Neurosciences Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><jtitle>Journal of cellular biochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>de-Freitas-Junior, Julio Cesar Madureira</au><au>Bastos, Lilian Golçalves</au><au>Freire-Neto, Carlos Alberto</au><au>Rocher, Bárbara Du</au><au>Abdelhay, Eliana Saul Furquim Werneck</au><au>Morgado-Díaz, José Andrés</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>N-glycan biosynthesis inhibitors induce in vitro anticancer activity in colorectal cancer cells</atitle><jtitle>Journal of cellular biochemistry</jtitle><addtitle>J. Cell. Biochem</addtitle><date>2012-09</date><risdate>2012</risdate><volume>113</volume><issue>9</issue><spage>2957</spage><epage>2966</epage><pages>2957-2966</pages><issn>0730-2312</issn><eissn>1097-4644</eissn><abstract>During malignant transformation, changes in the expression profile of glycans may be involved in a variety of events, including the loss of cell–cell and cell–matrix adhesion, migration, invasion, and evasion of apoptosis. Therefore, modulation of glycan expression with drugs has promising therapeutic potential for various cancer types. In this study, we investigated the in vitro anticancer activity of the N‐glycan biosynthesis inhibitors (swainsonine and tunicamycin) in cells derived from colorectal cancer (CRC). We also examined whether these inhibitors are able to induce radiosensitization and toxicity when used in combination with cisplatin or irinotecan, two current anticancer drugs. Our results show that treatment with tunicamycin inhibits cellular mechanisms related to the malignant phenotype, such as anchorage‐dependent and anchorage‐independent colony formation, migration and invasion, in undifferentiated HCT‐116 colon cancer cells, whereas swainsonine only inhibits cell migration. We also observed that tunicamycin, but not swainsonine, caused radiosensitivity in HCT‐116 cells. Moreover, the combination of swainsonine with cisplatin or irinotecan enhanced their toxicity in HCT‐116 cells, while the combination of tunicamycin with these drugs had no effect. Given these results, we suggest that the modulation of N‐glycan biosynthesis appears to be a potential therapeutic tool for CRC treatment because inhibition of this process induced anticancer activity in vitro. Additionally, the inhibition of the N‐glycan biosynthesis in combination with chemotherapic drugs is a promising therapeutic strategy for enhancing radiation therapy. J. Cell. Biochem. 113: 2957–2966, 2012. © 2012 Wiley Periodicals, Inc.</abstract><cop>Hoboken</cop><pub>Wiley Subscription Services, Inc., A Wiley Company</pub><pmid>22552949</pmid><doi>10.1002/jcb.24173</doi><tpages>10</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0730-2312 |
| ispartof | Journal of cellular biochemistry, 2012-09, Vol.113 (9), p.2957-2966 |
| issn | 0730-2312 1097-4644 |
| language | eng |
| recordid | cdi_proquest_journals_3113516599 |
| source | MEDLINE; Wiley Journals |
| subjects | Anchorages Anticancer properties Antineoplastic drugs Antitumor activity Apoptosis Biosynthesis Blotting, Western Caco-2 Cells Camptothecin - analogs & derivatives Camptothecin - pharmacology Cancer therapies Cell migration Cell Movement - drug effects Cell Survival - drug effects Cisplatin Cisplatin - pharmacology Colon cancer COLORECTAL CANCER Colorectal carcinoma Colorectal Neoplasms - metabolism Drugs Flow Cytometry Glycan HCT116 Cells HT29 Cells Humans Inhibitors Irinotecan Modulation N-GLYCOSYLATION Phenotypes Polysaccharides Polysaccharides - metabolism Radiation therapy RADIOSENSITIVITY Radiosensitization Swainsonine - pharmacology Toxicity TUNICAMYCIN Tunicamycin - pharmacology |
| title | N-glycan biosynthesis inhibitors induce in vitro anticancer activity in colorectal cancer cells |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-22T18%3A06%3A11IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=N-glycan%20biosynthesis%20inhibitors%20induce%20in%20vitro%20anticancer%20activity%20in%20colorectal%20cancer%20cells&rft.jtitle=Journal%20of%20cellular%20biochemistry&rft.au=de-Freitas-Junior,%20Julio%20Cesar%20Madureira&rft.date=2012-09&rft.volume=113&rft.issue=9&rft.spage=2957&rft.epage=2966&rft.pages=2957-2966&rft.issn=0730-2312&rft.eissn=1097-4644&rft_id=info:doi/10.1002/jcb.24173&rft_dat=%3Cproquest_cross%3E3113516599%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=3113516599&rft_id=info:pmid/22552949&rfr_iscdi=true |