P07-22 Particulate matter of Barranquilla (Colombia) affects reproduction and survival of Caenorhabditis elegans

Particulate matter is one of the main environmental issues in large cities due to traffic and industrial activity. Barranquilla is a city with more than 2 million habitants city with an intense port and industrial economy. In this work, we collected PM2.5 samples from the industrial area of Barranqu...

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Autores:
Arroyo, B.J.
Tejeda-Benitez, L.P.
Sierra-Marquez, L.I.
Moreno-Rios, A.L.
Godin-Arroyo, B.S.
Martinez-Avila, M.C.
Rodriguez Beltran, S.
Tipo de recurso:
Article of investigation
Fecha de publicación:
2022
Institución:
Corporación Universidad de la Costa
Repositorio:
REDICUC - Repositorio CUC
Idioma:
eng
OAI Identifier:
oai:repositorio.cuc.edu.co:11323/10799
Acceso en línea:
https://hdl.handle.net/11323/10799
https://repositorio.cuc.edu.co/
Palabra clave:
Caenorhabditis elegans
Reproduction
Survival
Rights
embargoedAccess
License
Atribución-NoComercial-SinDerivadas 4.0 Internacional (CC BY-NC-ND 4.0)
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dc.title.eng.fl_str_mv P07-22 Particulate matter of Barranquilla (Colombia) affects reproduction and survival of Caenorhabditis elegans
title P07-22 Particulate matter of Barranquilla (Colombia) affects reproduction and survival of Caenorhabditis elegans
spellingShingle P07-22 Particulate matter of Barranquilla (Colombia) affects reproduction and survival of Caenorhabditis elegans
Caenorhabditis elegans
Reproduction
Survival
title_short P07-22 Particulate matter of Barranquilla (Colombia) affects reproduction and survival of Caenorhabditis elegans
title_full P07-22 Particulate matter of Barranquilla (Colombia) affects reproduction and survival of Caenorhabditis elegans
title_fullStr P07-22 Particulate matter of Barranquilla (Colombia) affects reproduction and survival of Caenorhabditis elegans
title_full_unstemmed P07-22 Particulate matter of Barranquilla (Colombia) affects reproduction and survival of Caenorhabditis elegans
title_sort P07-22 Particulate matter of Barranquilla (Colombia) affects reproduction and survival of Caenorhabditis elegans
dc.creator.fl_str_mv Arroyo, B.J.
Tejeda-Benitez, L.P.
Sierra-Marquez, L.I.
Moreno-Rios, A.L.
Godin-Arroyo, B.S.
Martinez-Avila, M.C.
Rodriguez Beltran, S.
dc.contributor.author.none.fl_str_mv Arroyo, B.J.
Tejeda-Benitez, L.P.
Sierra-Marquez, L.I.
Moreno-Rios, A.L.
Godin-Arroyo, B.S.
Martinez-Avila, M.C.
Rodriguez Beltran, S.
dc.subject.proposal.eng.fl_str_mv Caenorhabditis elegans
Reproduction
Survival
topic Caenorhabditis elegans
Reproduction
Survival
description Particulate matter is one of the main environmental issues in large cities due to traffic and industrial activity. Barranquilla is a city with more than 2 million habitants city with an intense port and industrial economy. In this work, we collected PM2.5 samples from the industrial area of Barranquilla between July and September 2021, and obtained aqueous extracts from these samples. The nematode Caenorhabditis elegans was the toxicological model in this research. The wild type strain N2 was used for survival and reproduction assays. Previously, worms were subjected to age-synchronization via bleach solution. Experiments were performed using whole extract and dilutions of 50% and 33% to analyze dose dependence. For survival, about 10 nematodes in larval age L4 were exposed for 24 h to whole extract and dilutions. Four replicates for treatment were made and each experiment was conducted three. For reproduction, ten nematodes in larval stage L4 were exposed for 24 h to whole extract and dilutions. Each nematode was set individually in K agar-plates seeded with E. coli OP50, and the number of offspring at all stages was counted after 72 h. The results showed that the dust extracts to which the nematodes were exposed did not cause mortality in any dilution made, or with the complete extract. However, the extracts had sublethal effects that affected the reproduction of the model, by decreasing the number of eggs laid after 72 h of exposure to the dust extracts. This effect was concentration-dependent but did not vary with the month of collection, probably due to climate stability at this time of year in the Caribbean. This study is the first approximation of the toxicological and environmental behaviour of PM2.5 in the North of Colombia and verifies the usefulness of Caenorhabditis elegans as a biological model.
publishDate 2022
dc.date.issued.none.fl_str_mv 2022-09
dc.date.available.none.fl_str_mv 2023-09
2024-02-28T13:40:23Z
dc.date.accessioned.none.fl_str_mv 2024-02-28T13:40:23Z
dc.type.spa.fl_str_mv Artículo de revista
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dc.identifier.citation.spa.fl_str_mv B.J. Arroyo, L.P. Tejeda-Benitez, L.I. Sierra-Marquez, A.L. Moreno-Rios, B.S. Godin-Arroyo, M.C. Martinez-Avila, S. Rodriguez Beltran, P07-22 Particulate matter of Barranquilla (Colombia) affects reproduction and survival of Caenorhabditis elegans, Toxicology Letters, Volume 368, Supplement, 2022, Page S128, ISSN 0378-4274, https://doi.org/10.1016/j.toxlet.2022.07.365
dc.identifier.issn.spa.fl_str_mv 0378-4274
dc.identifier.uri.none.fl_str_mv https://hdl.handle.net/11323/10799
dc.identifier.doi.none.fl_str_mv 10.1016/j.toxlet.2022.07.365
dc.identifier.eissn.spa.fl_str_mv 1879-3169
dc.identifier.instname.spa.fl_str_mv Corporación Universidad de la Costa
dc.identifier.reponame.spa.fl_str_mv REDICUC – Repositorio CUC
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identifier_str_mv B.J. Arroyo, L.P. Tejeda-Benitez, L.I. Sierra-Marquez, A.L. Moreno-Rios, B.S. Godin-Arroyo, M.C. Martinez-Avila, S. Rodriguez Beltran, P07-22 Particulate matter of Barranquilla (Colombia) affects reproduction and survival of Caenorhabditis elegans, Toxicology Letters, Volume 368, Supplement, 2022, Page S128, ISSN 0378-4274, https://doi.org/10.1016/j.toxlet.2022.07.365
0378-4274
10.1016/j.toxlet.2022.07.365
1879-3169
Corporación Universidad de la Costa
REDICUC – Repositorio CUC
url https://hdl.handle.net/11323/10799
https://repositorio.cuc.edu.co/
dc.language.iso.spa.fl_str_mv eng
language eng
dc.relation.ispartofjournal.spa.fl_str_mv Toxicology Letters
dc.relation.references.spa.fl_str_mv 1 M García-Espiñeira, L Tejeda-Benitez, J Olivero-Verbel Toxicity of atrazine-and glyphosate-based formulations on Caenorhabditis elegans Ecotoxicol Environ Saf (2018), pp. 216-222
2 L Tejeda-Benítez, K Noguera-Oviedo, DS Aga, J Olivero-Verbel Toxicity profile of organic extracts from Magdalena River sediments Environ Sci Pollut Res Int (2018), pp. 519-1532
3 AL Moreno-Ríos, LP Tejeda-Benítez, CF Bustillo-Lecompte Sources, characteristics, toxicity, and control of ultrafine particles: An overview Geoscience Frontiers (2021), p. 101147
4 XN Ma, RQ Li, JL Xie, SH Li, JW Li, XX Yan PM2.5-induced inflammation and myocardial cell injury in rats Eur Rev Med Pharmacol Sci (2021), pp. 6670-6677
5 ZL Jia, CY Zhu, RS Rajendran, Q Xia, KC Liu, Y Zhang Impact of airborne total suspended particles (TSP) and fine particulate matter (PM2.5)-induced developmental toxicity in zebrafish (Danio rerio) embryos J Appl Toxicol (2022)
dc.relation.citationstartpage.spa.fl_str_mv S128
dc.relation.citationissue.spa.fl_str_mv 1
dc.relation.citationvolume.spa.fl_str_mv 368
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dc.coverage.city.none.fl_str_mv Barranquilla
dc.coverage.country.none.fl_str_mv Colombia
dc.publisher.spa.fl_str_mv Elsevier BV
dc.publisher.place.spa.fl_str_mv Netherlands
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spelling Atribución-NoComercial-SinDerivadas 4.0 Internacional (CC BY-NC-ND 4.0)https://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/embargoedAccesshttp://purl.org/coar/access_right/c_f1cfArroyo, B.J.Tejeda-Benitez, L.P.Sierra-Marquez, L.I.Moreno-Rios, A.L.Godin-Arroyo, B.S.Martinez-Avila, M.C.Rodriguez Beltran, S.2024-02-28T13:40:23Z2023-092024-02-28T13:40:23Z2022-09B.J. Arroyo, L.P. Tejeda-Benitez, L.I. Sierra-Marquez, A.L. Moreno-Rios, B.S. Godin-Arroyo, M.C. Martinez-Avila, S. Rodriguez Beltran, P07-22 Particulate matter of Barranquilla (Colombia) affects reproduction and survival of Caenorhabditis elegans, Toxicology Letters, Volume 368, Supplement, 2022, Page S128, ISSN 0378-4274, https://doi.org/10.1016/j.toxlet.2022.07.3650378-4274https://hdl.handle.net/11323/1079910.1016/j.toxlet.2022.07.3651879-3169Corporación Universidad de la CostaREDICUC – Repositorio CUChttps://repositorio.cuc.edu.co/Particulate matter is one of the main environmental issues in large cities due to traffic and industrial activity. Barranquilla is a city with more than 2 million habitants city with an intense port and industrial economy. In this work, we collected PM2.5 samples from the industrial area of Barranquilla between July and September 2021, and obtained aqueous extracts from these samples. The nematode Caenorhabditis elegans was the toxicological model in this research. The wild type strain N2 was used for survival and reproduction assays. Previously, worms were subjected to age-synchronization via bleach solution. Experiments were performed using whole extract and dilutions of 50% and 33% to analyze dose dependence. For survival, about 10 nematodes in larval age L4 were exposed for 24 h to whole extract and dilutions. Four replicates for treatment were made and each experiment was conducted three. For reproduction, ten nematodes in larval stage L4 were exposed for 24 h to whole extract and dilutions. Each nematode was set individually in K agar-plates seeded with E. coli OP50, and the number of offspring at all stages was counted after 72 h. The results showed that the dust extracts to which the nematodes were exposed did not cause mortality in any dilution made, or with the complete extract. However, the extracts had sublethal effects that affected the reproduction of the model, by decreasing the number of eggs laid after 72 h of exposure to the dust extracts. This effect was concentration-dependent but did not vary with the month of collection, probably due to climate stability at this time of year in the Caribbean. This study is the first approximation of the toxicological and environmental behaviour of PM2.5 in the North of Colombia and verifies the usefulness of Caenorhabditis elegans as a biological model.1 páginaapplication/pdfengElsevier BVNetherlandshttps://www-sciencedirect-com.ezproxy.cuc.edu.co/science/article/pii/S0378427422013261?via%3DihubP07-22 Particulate matter of Barranquilla (Colombia) affects reproduction and survival of Caenorhabditis elegansArtículo de revistahttp://purl.org/coar/resource_type/c_2df8fbb1Textinfo:eu-repo/semantics/articlehttp://purl.org/redcol/resource_type/ARTinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/version/c_970fb48d4fbd8a85BarranquillaColombiaToxicology Letters1 M García-Espiñeira, L Tejeda-Benitez, J Olivero-Verbel Toxicity of atrazine-and glyphosate-based formulations on Caenorhabditis elegans Ecotoxicol Environ Saf (2018), pp. 216-2222 L Tejeda-Benítez, K Noguera-Oviedo, DS Aga, J Olivero-Verbel Toxicity profile of organic extracts from Magdalena River sediments Environ Sci Pollut Res Int (2018), pp. 519-15323 AL Moreno-Ríos, LP Tejeda-Benítez, CF Bustillo-Lecompte Sources, characteristics, toxicity, and control of ultrafine particles: An overview Geoscience Frontiers (2021), p. 1011474 XN Ma, RQ Li, JL Xie, SH Li, JW Li, XX Yan PM2.5-induced inflammation and myocardial cell injury in rats Eur Rev Med Pharmacol Sci (2021), pp. 6670-66775 ZL Jia, CY Zhu, RS Rajendran, Q Xia, KC Liu, Y Zhang Impact of airborne total suspended particles (TSP) and fine particulate matter (PM2.5)-induced developmental toxicity in zebrafish (Danio rerio) embryos J Appl Toxicol (2022)S1281368Caenorhabditis elegansReproductionSurvivalPublicationORIGINALP07-22 Particulate matter of Barranquilla (Colombia) affects reproduction and survival of Caenorhabditis elegans.pdfP07-22 Particulate matter of Barranquilla (Colombia) affects reproduction and survival of Caenorhabditis 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ada en las Obras Colectivas.

b.	Distribuir copias o fonogramas de las Obras, exhibirlas públicamente, ejecutarlas públicamente y/o ponerlas a disposición pública, incluyéndolas como incorporadas en Obras Colectivas, según corresponda.

c.	Distribuir copias de las Obras Derivadas que se generen, exhibirlas públicamente, ejecutarlas públicamente y/o ponerlas a disposición pública.
Los derechos mencionados anteriormente pueden ser ejercidos en todos los medios y formatos, actualmente conocidos o que se inventen en el futuro. Los derechos antes mencionados incluyen el derecho a realizar dichas modificaciones en la medida que sean técnicamente necesarias para ejercer los derechos en otro medio o formatos, pero de otra manera usted no está autorizado para realizar obras derivadas. Todos los derechos no otorgados expresamente por el Licenciante quedan por este medio reservados, incluyendo pero sin limitarse a aquellos que se mencionan en las secciones 4(d) y 4(e).

4. Restricciones.
La licencia otorgada en la anterior Sección 3 está expresamente sujeta y limitada por las siguientes restricciones:

a.	Usted puede distribuir, exhibir públicamente, ejecutar públicamente, o poner a disposición pública la Obra sólo bajo las condiciones de esta Licencia, y Usted debe incluir una copia de esta licencia o del Identificador Universal de Recursos de la misma con cada copia de la Obra que distribuya, exhiba públicamente, ejecute públicamente o ponga a disposición pública. No es posible ofrecer o imponer ninguna condición sobre la Obra que altere o limite las condiciones de esta Licencia o el ejercicio de los derechos de los destinatarios otorgados en este documento. No es posible sublicenciar la Obra. Usted debe mantener intactos todos los avisos que hagan referencia a esta Licencia y a la cláusula de limitación de garantías. Usted no puede distribuir, exhibir públicamente, ejecutar públicamente, o poner a disposición pública la Obra con alguna medida tecnológica que controle el acceso o la utilización de ella de una forma que sea inconsistente con las condiciones de esta Licencia. Lo anterior se aplica a la Obra incorporada a una Obra Colectiva, pero esto no exige que la Obra Colectiva aparte de la obra misma quede sujeta a las condiciones de esta Licencia. Si Usted crea una Obra Colectiva, previo aviso de cualquier Licenciante debe, en la medida de lo posible, eliminar de la Obra Colectiva cualquier referencia a dicho Licenciante o al Autor Original, según lo solicitado por el Licenciante y conforme lo exige la cláusula 4(c).

b.	Usted no puede ejercer ninguno de los derechos que le han sido otorgados en la Sección 3 precedente de modo que estén principalmente destinados o directamente dirigidos a conseguir un provecho comercial o una compensación monetaria privada. El intercambio de la Obra por otras obras protegidas por derechos de autor, ya sea a través de un sistema para compartir archivos digitales (digital file-sharing) o de cualquier otra manera no será considerado como estar destinado principalmente o dirigido directamente a conseguir un provecho comercial o una compensación monetaria privada, siempre que no se realice un pago mediante una compensación monetaria en relación con el intercambio de obras protegidas por el derecho de autor.

c.	Si usted distribuye, exhibe públicamente, ejecuta públicamente o ejecuta públicamente en forma digital la Obra o cualquier Obra Derivada u Obra Colectiva, Usted debe mantener intacta toda la información de derecho de autor de la Obra y proporcionar, de forma razonable según el medio o manera que Usted esté utilizando: (i) el nombre del Autor Original si está provisto (o seudónimo, si fuere aplicable), y/o (ii) el nombre de la parte o las partes que el Autor Original y/o el Licenciante hubieren designado para la atribución (v.g., un instituto patrocinador, editorial, publicación) en la información de los derechos de autor del Licenciante, términos de servicios o de otras formas razonables; el título de la Obra si está provisto; en la medida de lo razonablemente factible y, si está provisto, el Identificador Uniforme de Recursos (Uniform Resource Identifier) que el Licenciante especifica para ser asociado con la Obra, salvo que tal URI no se refiera a la nota sobre los derechos de autor o a la información sobre el licenciamiento de la Obra; y en el caso de una Obra Derivada, atribuir el crédito identificando el uso de la Obra en la Obra Derivada (v.g., "Traducción Francesa de la Obra del Autor Original," o "Guión Cinematográfico basado en la Obra original del Autor Original"). Tal crédito puede ser implementado de cualquier forma razonable; en el caso, sin embargo, de Obras Derivadas u Obras Colectivas, tal crédito aparecerá, como mínimo, donde aparece el crédito de cualquier otro autor comparable y de una manera, al menos, tan destacada como el crédito de otro autor comparable.

d.	Para evitar toda confusión, el Licenciante aclara que, cuando la obra es una composición musical:

i.	Regalías por interpretación y ejecución bajo licencias generales. El Licenciante se reserva el derecho exclusivo de autorizar la ejecución pública o la ejecución pública digital de la obra y de recolectar, sea individualmente o a través de una sociedad de gestión colectiva de derechos de autor y derechos conexos (por ejemplo, SAYCO), las regalías por la ejecución pública o por la ejecución pública digital de la obra (por ejemplo Webcast) licenciada bajo licencias generales, si la interpretación o ejecución de la obra está primordialmente orientada por o dirigida a la obtención de una ventaja comercial o una compensación monetaria privada.

ii.	Regalías por Fonogramas. El Licenciante se reserva el derecho exclusivo de recolectar, individualmente o a través de una sociedad de gestión colectiva de derechos de autor y derechos conexos (por ejemplo, los consagrados por la SAYCO), una agencia de derechos musicales o algún agente designado, las regalías por cualquier fonograma que Usted cree a partir de la obra (“versión cover”) y distribuya, en los términos del régimen de derechos de autor, si la creación o distribución de esa versión cover está primordialmente destinada o dirigida a obtener una ventaja comercial o una compensación monetaria privada.

e.	Gestión de Derechos de Autor sobre Interpretaciones y Ejecuciones Digitales (WebCasting). Para evitar toda confusión, el Licenciante aclara que, cuando la obra sea un fonograma, el Licenciante se reserva el derecho exclusivo de autorizar la ejecución pública digital de la obra (por ejemplo, webcast) y de recolectar, individualmente o a través de una sociedad de gestión colectiva de derechos de autor y derechos conexos (por ejemplo, ACINPRO), las regalías por la ejecución pública digital de la obra (por ejemplo, webcast), sujeta a las disposiciones aplicables del régimen de Derecho de Autor, si esta ejecución pública digital está primordialmente dirigida a obtener una ventaja comercial o una compensación monetaria privada.

5. Representaciones, Garantías y Limitaciones de Responsabilidad.
A MENOS QUE LAS PARTES LO ACORDARAN DE OTRA FORMA POR ESCRITO, EL LICENCIANTE OFRECE LA OBRA (EN EL ESTADO EN EL QUE SE ENCUENTRA) “TAL CUAL”, SIN BRINDAR GARANTÍAS DE CLASE ALGUNA RESPECTO DE LA OBRA, YA SEA EXPRESA, IMPLÍCITA, LEGAL O CUALQUIERA OTRA, INCLUYENDO, SIN LIMITARSE A ELLAS, GARANTÍAS DE TITULARIDAD, COMERCIABILIDAD, ADAPTABILIDAD O ADECUACIÓN A PROPÓSITO DETERMINADO, AUSENCIA DE INFRACCIÓN, DE AUSENCIA DE DEFECTOS LATENTES O DE OTRO TIPO, O LA PRESENCIA O AUSENCIA DE ERRORES, SEAN O NO DESCUBRIBLES (PUEDAN O NO SER ESTOS DESCUBIERTOS). ALGUNAS JURISDICCIONES NO PERMITEN LA EXCLUSIÓN DE GARANTÍAS IMPLÍCITAS, EN CUYO CASO ESTA EXCLUSIÓN PUEDE NO APLICARSE A USTED.

6. Limitación de responsabilidad.
A MENOS QUE LO EXIJA EXPRESAMENTE LA LEY APLICABLE, EL LICENCIANTE NO SERÁ RESPONSABLE ANTE USTED POR DAÑO ALGUNO, SEA POR RESPONSABILIDAD EXTRACONTRACTUAL, PRECONTRACTUAL O CONTRACTUAL, OBJETIVA O SUBJETIVA, SE TRATE DE DAÑOS MORALES O PATRIMONIALES, DIRECTOS O INDIRECTOS, PREVISTOS O IMPREVISTOS PRODUCIDOS POR EL USO DE ESTA LICENCIA O DE LA OBRA, AUN CUANDO EL LICENCIANTE HAYA SIDO ADVERTIDO DE LA POSIBILIDAD DE DICHOS DAÑOS. ALGUNAS LEYES NO PERMITEN LA EXCLUSIÓN DE CIERTA RESPONSABILIDAD, EN CUYO CASO ESTA EXCLUSIÓN PUEDE NO APLICARSE A USTED.

7. Término.

a.	Esta Licencia y los derechos otorgados en virtud de ella terminarán automáticamente si Usted infringe alguna condición establecida en ella. Sin embargo, los individuos o entidades que han recibido Obras Derivadas o Colectivas de Usted de conformidad con esta Licencia, no verán terminadas sus licencias, siempre que estos individuos o entidades sigan cumpliendo íntegramente las condiciones de estas licencias. Las Secciones 1, 2, 5, 6, 7, y 8 subsistirán a cualquier terminación de esta Licencia.

b.	Sujeta a las condiciones y términos anteriores, la licencia otorgada aquí es perpetua (durante el período de vigencia de los derechos de autor de la obra). No obstante lo anterior, el Licenciante se reserva el derecho a publicar y/o estrenar la Obra bajo condiciones de licencia diferentes o a dejar de distribuirla en los términos de esta Licencia en cualquier momento; en el entendido, sin embargo, que esa elección no servirá para revocar esta licencia o que deba ser otorgada , bajo los términos de esta licencia), y esta licencia continuará en pleno vigor y efecto a menos que sea terminada como se expresa atrás. La Licencia revocada continuará siendo plenamente vigente y efectiva si no se le da término en las condiciones indicadas anteriormente.

8. Varios.

a.	Cada vez que Usted distribuya o ponga a disposición pública la Obra o una Obra Colectiva, el Licenciante ofrecerá al destinatario una licencia en los mismos términos y condiciones que la licencia otorgada a Usted bajo esta Licencia.

b.	Si alguna disposición de esta Licencia resulta invalidada o no exigible, según la legislación vigente, esto no afectará ni la validez ni la aplicabilidad del resto de condiciones de esta Licencia y, sin acción adicional por parte de los sujetos de este acuerdo, aquélla se entenderá reformada lo mínimo necesario para hacer que dicha disposición sea válida y exigible.

c.	Ningún término o disposición de esta Licencia se estimará renunciada y ninguna violación de ella será consentida a menos que esa renuncia o consentimiento sea otorgado por escrito y firmado por la parte que renuncie o consienta.

d.	Esta Licencia refleja el acuerdo pleno entre las partes respecto a la Obra aquí licenciada. No hay arreglos, acuerdos o declaraciones respecto a la Obra que no estén especificados en este documento. El Licenciante no se verá limitado por ninguna disposición adicional que pueda surgir en alguna comunicación emanada de Usted. Esta Licencia no puede ser modificada sin el consentimiento mutuo por escrito del Licenciante y Usted.
