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42. | | BUZZATO, V. C.; OLIVEIRA, A. L. de; ROCHA, M. I. P.; MELLO, S. S. de; NICIURA, S. C. M. Efeitos de sistemas nanoestruturados sobre a expressão gênica em fibroblastos bovinos in vitro In: JORNADA CIENTÍFICA - EMBRAPA SÃO CARLOS, 5., 2013, São Carlos, SP. Anais... São Carlos, SP: Embrapa Pecuária Sudeste: Embrapa Instrumentação , 2013. p. 11 (Embrapa Pecuária Sudeste. Documentos, 110). Editado por Ana Rita de Araújo Nogueira, Simone Cristina Méo Niciura Biblioteca(s): Embrapa Pecuária Sudeste. |
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43. | | MELLO, S. C.; ROCHA, M. I. P.; GROMBONI, J. G. G.; VENERONI, G. B.; NICIURA, S. C. M. Descrição de novos polimorfismos do tipo SNP no gene da calpastatina em bovinos. In: CONGRESSO BRASILEIRO DE GENÉTICA, 56., 2010, Guarujá, SP. Resumos... Guarujá: Sociedade Brasileira de Genética, 2010. Biblioteca(s): Embrapa Pecuária Sudeste. |
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44. | | ROCHA, M. I. P.; MELLO, S. de C.; GROMBONI, J. G. G.; OTSUK, I. P.; NICIURA, S. C. M. Frequência do polimorfismo F200Y no gene da b-tubulina em em rebanhos ovinos do Estado de São Paulo. In: JORNADA CIENTÍFICA - EMBRAPA SÃO CARLOS, 2., 2010, São Carlos, SP. Anais... São Carlos: Embrapa Instrumentação Agropecuária: Embrapa Pecuária Sudeste, 2010. Biblioteca(s): Embrapa Pecuária Sudeste. |
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47. | | NUCCI, A. DA SILVA; GIRALDELO, L. A.; SANTOS, I. B. DOS; ALEMÁN GAINZA, Y.; NICIURA, S. C. M. Padronização de co-cultivo in vitro de Haemonchus contortus com células abomasais ovinas. In: JORNADA CIENTÍFICA DA EMBRAPA SÃO CARLOS, 12., 2020, São Carlos, SP. Anais... São Carlos, SP: Embrapa Instrumentação; Embrapa Pecuária Sudeste, 2020. p.13. (Embrapa Instrumentação. Documentos, 71). Biblioteca(s): Embrapa Pecuária Sudeste. |
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51. | | SARAIVA, N. Z.; OLIVEIRA, C. S.; TETZNER, T. A. D.; LIMA, M. R. de; MELO, D. S. de; NICIURA, S. C. M.; GARCIA, J. M. Chemically assisted enucleation results in higher G6PD expression in early bovine female embryos obtained by somatic cell nuclear transfer. Cellular Reprogramming, v. 14, n. 5, p. 1-11, 2012. Biblioteca(s): Embrapa Pecuária Sudeste. |
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52. | | SARAIVA, N. Z.; OLIVEIRA, C. S.; LEAL, C. L. V.; CALLADO, M. del; VANTINI, R.; MONTEIRO, F. M.; NICIURA, S. C. M.; GARCIA, J. M. Chemically induced enucleation of activated bovine oocytes: chromatin and microtubule organization and production of viable cytoplasts. Zygote, v. 23, n. 6, p. 852-862, 2015. Biblioteca(s): Embrapa Gado de Leite; Embrapa Pecuária Sudeste. |
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53. | | BRESSANI, F. A.; TIZIOTO, P. C.; ROCHA, M. I. P.; IBELLI, A. M. G.; NICIURA, S. C. M.; REGITANO, L. C. de A. Comparação de diferentes protocolos para visualização de DNA separado por eletroforese em gel de agarose utilizando-se os corantes fluorescentes brometo de etídeo e GelRedr. In: ENCONTRO NACIONAL SOBRE MÉTODOS DOS LABORATÓRIOS DA EMBRAPA, 15., 2010, Pelotas, RS. Novas perspectivas para os laboratórios da Embrapa. Pelotas: Embrapa Clima Temperado, 2010. Biblioteca(s): Embrapa Pecuária Sudeste. |
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54. | | BRAZ, F. S. S.; CRUZ, A. A.; PAIS, F.; OLIVEIRA, G.; NICIURA, S. C. M.; BEECH, R.; MOLENTO, M. B.; PRICHARD, R. SNPS/INDELS identification in genomic regions of abc transporters in susceptible and ivermectin-resistant Haemonchus contortus In: CONGRESSO BRASILEIRO DE PARASITOLOGIA VETERINÁRIA, 18., 2014, Gramado. Anais... Gramado: CBPV, 2014. Biblioteca(s): Embrapa Pecuária Sudeste. |
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55. | | NICIURA, S. C. M.; ROCHA, M. I. P.; CRUVINEL, G. G.; FERRACIOLI, C. de M.; ESTEVES, S. N.; CHAGAS, A. C. de S. In silico evidence of epigenetic machinery proteins in the nematode Haemonchus contortus. In: MOLECULAR HELMINTHOLOGY: AN INTEGRATED APPROACH, 1., Cape Cod, MA, USA. Proceedings... Cape Cod, MA, USA: Elsevier, 2017. p. 29. Biblioteca(s): Embrapa Pecuária Sudeste. |
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56. | | CHAGAS, A. C. de S.; DOMINGUES, L. F.; ALEMÁN GAINZA, Y.; BARIONI JUNIOR, W.; ESTEVES, S. N.; NICIURA, S. C. M. Target selected treatment with levamisole to control the development of anthelmintic resistance in a sheep flock. Parasitology Research, v. 115, n. 3, p. 1131-1139, mar. 2016. Biblioteca(s): Embrapa Pecuária Sudeste. |
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57. | | NICIURA, S. C. M.; BENAVIDES, M. V.; OKINO, C. H.; IBELLI, A. M. G.; MINHO, A. P.; ESTEVES, S. N.; CHAGAS, A. C. de S. Genome-wide association study for Haemonchus contortus resistance in Morada Nova sheep. Pathogens, v. 11, n. 8, aug. 2022, 939. 12 p. Biblioteca(s): Embrapa Pecuária Sudeste; Embrapa Pecuária Sul; Embrapa Suínos e Aves. |
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58. | | DEA, R. C. D.; CATOIA, V.; TIZIOTO, P. C.; ROCHA, M. I. P.; REGITANO, L. C. de A.; NICIURA, S. C. M. Genotipagem de SNPs em genes candidatos a características de interesse comercial em bovinos de corte com sondas de hidrólise. In: JORNADA CIENTÍFICA - EMBRAPA SÃO CARLOS, 4., 2012, São Carlos, SP. Anais... São Carlos: Embrapa Instrumentação: Embrapa Pecuária Sudeste, 2012. p. 32. (Embrapa Instrumentação. Documentos, 56). Biblioteca(s): Embrapa Pecuária Sudeste. |
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59. | | GROMBONI, J. G. G.; GAGLIARDI, T. R.; MELLO, S. S.; GIUSTI, J.; TURIM, E.; VERÍSSIMO, C. J.; MOLENTO, M. B.; NICIURA, S. C. M. Identificação molecular do helminto Haemonchus contortus e de polimorfismo no gene da b-tubulina. In: JORNADA CIENTÍFICA-EMBRAPA SÃO CARLOS, 2009, São Carlos, SP. Anais... São Carlos: Embrapa Pecuária Sudeste: Embrapa Instrumentação Agropecuária, 2009. Editado por Luiz Francisco Zafalon, Simone Cristina Méo Niciura. Biblioteca(s): Embrapa Pecuária Sudeste. |
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60. | | MELLO, S. S. de; ROCHA, M. I. P.; GIGLIOTI, R.; BRESSANI, F. A. B.; MALAGO JUNIOR, W.; NICIURA, S. C. M. Identificação de polimorfismos no gene da glicoproteína-P (PgP) em Haemonchus contortus: dificuldades técnicas. In: JORNADA CIENTÍFICA - EMBRAPA SÃO CARLOS, 4., 2012, São Carlos, SP. Anais... São Carlos: Embrapa Instrumentação: Embrapa Pecuária Sudeste, 2012. p. 48. (Embrapa Instrumentação. Documentos, 56). Biblioteca(s): Embrapa Pecuária Sudeste. |
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Registros recuperados : 173 | |
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Registro Completo
Biblioteca(s): |
Embrapa Pecuária Sudeste. |
Data corrente: |
28/09/2010 |
Data da última atualização: |
30/06/2023 |
Tipo da produção científica: |
Resumo em Anais de Congresso |
Autoria: |
SARAIVA, N. Z.; OLIVEIRA, C. S.; TETZNER, T. A. D.; SOUZA, M. M.; LIMA, M. R. de; NICIURA, S. C. M.; GARCIA, J. M. |
Afiliação: |
N. Z. SARAIVA, UNESP/JABOTICABAL; C. S. OLIVEIRA, UNESP/JABOTICABAL; T. A. D. TETZNER, UNESP/JABOTICABAL; M. M. SOUZA, UNESP/JABOTICABAL; M. R. DE LIMA, UNESP/JABOTICABAL; SIMONE CRISTINA MEO NICIURA, CPPSE; J. M. GARCIA, UNESP/JABOTICABAL. |
Título: |
Bovine cytoplasts prepared by demecolcine-induced enucleation of activated oocytes. |
Ano de publicação: |
2010 |
Fonte/Imprenta: |
In: ANNUAL CONFERENCE OT THE INTERNATIONAL EMBRYO TRANSFER SOCIETY, 36., 2010, Cordoba. Proceedings... Córdoba: ITES, 2010; Reproduction, Fertility and development, v. 22, n. 1, 2010. |
Páginas: |
p. 197 |
Idioma: |
Português |
Conteúdo: |
One of the most critical steps of the standard NT procedure is the removal of oocyte chromosomes for the production of enucleated cytoplasts. This process involves ultraviolet (UV) irradiation, which causes damage to the membrane and intracellular components of bovine oocytes. The objective of the present study was to evaluate the use of demecolcine, a microtubule depolymerizing agent, for chemical-induced enucleation of activated bovine oocytes. In the first experiment, oocytes obtained from cow ovaries collected in a slaughterhouse were IVM in TCM-199 medium supplemented with 10% FCS, FSH, hCG, estradiol, pyruvate, and amikacin for 26 h; then, they were artificially activated with 5 ?M ionomycin for 5 min and 10 ?g mL-1 cycloheximide for 4 h. The following groups were established: control, C (no activation and no demecolcine); activated, A (exposure only to activating agents); and demecolcine-exposed oocytes, DEME (activation for 4 h and 0.05 ?g mL-1 demecolcine for 2 to 4 h after 0, 0.5, 1.0, 1.5, and 2.0 h of activation). The treatment DEME comprised the groups G1 (demecolcine from 0 to 2 h of activation; DEME 0-2 h); G2 (DEME 0-4 h); G3 (DEME 0.5-2.5 h); G4 (DEME 0.5-4 h); G5 (DEME 1-3 h); G6 (DEME 1-4 h); G7 (DEME 1.5-3.5 h); G8 (DEME 1.5-4 h); and G9 (DEME 2-4h). The oocytes were stained with 10 ?g mL-1 Hoechst 33342 for 15 min and enucleation rates (EN) were evaluated under epifluorescence microscope (330-385 nm).After EN evaluation, a second experiment was performed to verify the efficiency of demecolcine-induced enucleation in group G9, which showed the best result in the first experiment. After demecolcine treatment, the oocytes were incubated in HEPES-buffered SOF with 10% FCS and cytochalasin B for removal of the 2PB and minimal adjacent cytoplast under an inverted optical microscope. Traditional enucleation was performed on the control group without exposure of the oocytes to demecolcine. The same conditions were employed except that UV light was used to confirm enucleation. Samples of cytoplasts were stained with Hoechst for 15 min and analyzed for enucleation efficiency. Five and 3 replicates were evaluated, respectively, in experiments 1 and 2, and the results were analyzed by chi-square test in the statistical software Minitab®, release 14.1 (Minitab Inc., State College, PA, USA). A level of 5% significance was used. Regarding enucleation rates, all treated groups were significantly different compared with the C (0/114) and A (0/130) groups. Considering the DEME groups, G8 (46/109; 42.2%) and G9 (61/113; 54.0%) presented superior rates (P < 0.05) to all other groups (26.8 to 36.3%), but they were similar despite the great tendency (P = 0.07) to difference. We observed high efficiency (P < 0.05) of demecolcine-induced enucleation (90/110; 81.8%) compared with a traditional technique (61/95; 64.2%). In conclusion, the present study shows that demecolcine-induced enucleation of activated oocytes enables good rates of enucleation and has greater efficiency than the traditional technique as well as avoiding UV irradiation of the cytoplast. MenosOne of the most critical steps of the standard NT procedure is the removal of oocyte chromosomes for the production of enucleated cytoplasts. This process involves ultraviolet (UV) irradiation, which causes damage to the membrane and intracellular components of bovine oocytes. The objective of the present study was to evaluate the use of demecolcine, a microtubule depolymerizing agent, for chemical-induced enucleation of activated bovine oocytes. In the first experiment, oocytes obtained from cow ovaries collected in a slaughterhouse were IVM in TCM-199 medium supplemented with 10% FCS, FSH, hCG, estradiol, pyruvate, and amikacin for 26 h; then, they were artificially activated with 5 ?M ionomycin for 5 min and 10 ?g mL-1 cycloheximide for 4 h. The following groups were established: control, C (no activation and no demecolcine); activated, A (exposure only to activating agents); and demecolcine-exposed oocytes, DEME (activation for 4 h and 0.05 ?g mL-1 demecolcine for 2 to 4 h after 0, 0.5, 1.0, 1.5, and 2.0 h of activation). The treatment DEME comprised the groups G1 (demecolcine from 0 to 2 h of activation; DEME 0-2 h); G2 (DEME 0-4 h); G3 (DEME 0.5-2.5 h); G4 (DEME 0.5-4 h); G5 (DEME 1-3 h); G6 (DEME 1-4 h); G7 (DEME 1.5-3.5 h); G8 (DEME 1.5-4 h); and G9 (DEME 2-4h). The oocytes were stained with 10 ?g mL-1 Hoechst 33342 for 15 min and enucleation rates (EN) were evaluated under epifluorescence microscope (330-385 nm).After EN evaluation, a second experiment was performed... Mostrar Tudo |
Palavras-Chave: |
Bovine; Cytoplasts. |
Thesaurus NAL: |
oocytes. |
Categoria do assunto: |
W Química e Física |
Marc: |
LEADER 03893nam a2200229 a 4500 001 1863081 005 2023-06-30 008 2010 bl uuuu u00u1 u #d 100 1 $aSARAIVA, N. Z. 245 $aBovine cytoplasts prepared by demecolcine-induced enucleation of activated oocytes.$h[electronic resource] 260 $aIn: ANNUAL CONFERENCE OT THE INTERNATIONAL EMBRYO TRANSFER SOCIETY, 36., 2010, Cordoba. Proceedings... Córdoba: ITES, 2010; Reproduction, Fertility and development, v. 22, n. 1$c2010 300 $ap. 197 520 $aOne of the most critical steps of the standard NT procedure is the removal of oocyte chromosomes for the production of enucleated cytoplasts. This process involves ultraviolet (UV) irradiation, which causes damage to the membrane and intracellular components of bovine oocytes. The objective of the present study was to evaluate the use of demecolcine, a microtubule depolymerizing agent, for chemical-induced enucleation of activated bovine oocytes. In the first experiment, oocytes obtained from cow ovaries collected in a slaughterhouse were IVM in TCM-199 medium supplemented with 10% FCS, FSH, hCG, estradiol, pyruvate, and amikacin for 26 h; then, they were artificially activated with 5 ?M ionomycin for 5 min and 10 ?g mL-1 cycloheximide for 4 h. The following groups were established: control, C (no activation and no demecolcine); activated, A (exposure only to activating agents); and demecolcine-exposed oocytes, DEME (activation for 4 h and 0.05 ?g mL-1 demecolcine for 2 to 4 h after 0, 0.5, 1.0, 1.5, and 2.0 h of activation). The treatment DEME comprised the groups G1 (demecolcine from 0 to 2 h of activation; DEME 0-2 h); G2 (DEME 0-4 h); G3 (DEME 0.5-2.5 h); G4 (DEME 0.5-4 h); G5 (DEME 1-3 h); G6 (DEME 1-4 h); G7 (DEME 1.5-3.5 h); G8 (DEME 1.5-4 h); and G9 (DEME 2-4h). The oocytes were stained with 10 ?g mL-1 Hoechst 33342 for 15 min and enucleation rates (EN) were evaluated under epifluorescence microscope (330-385 nm).After EN evaluation, a second experiment was performed to verify the efficiency of demecolcine-induced enucleation in group G9, which showed the best result in the first experiment. After demecolcine treatment, the oocytes were incubated in HEPES-buffered SOF with 10% FCS and cytochalasin B for removal of the 2PB and minimal adjacent cytoplast under an inverted optical microscope. Traditional enucleation was performed on the control group without exposure of the oocytes to demecolcine. The same conditions were employed except that UV light was used to confirm enucleation. Samples of cytoplasts were stained with Hoechst for 15 min and analyzed for enucleation efficiency. Five and 3 replicates were evaluated, respectively, in experiments 1 and 2, and the results were analyzed by chi-square test in the statistical software Minitab®, release 14.1 (Minitab Inc., State College, PA, USA). A level of 5% significance was used. Regarding enucleation rates, all treated groups were significantly different compared with the C (0/114) and A (0/130) groups. Considering the DEME groups, G8 (46/109; 42.2%) and G9 (61/113; 54.0%) presented superior rates (P < 0.05) to all other groups (26.8 to 36.3%), but they were similar despite the great tendency (P = 0.07) to difference. We observed high efficiency (P < 0.05) of demecolcine-induced enucleation (90/110; 81.8%) compared with a traditional technique (61/95; 64.2%). In conclusion, the present study shows that demecolcine-induced enucleation of activated oocytes enables good rates of enucleation and has greater efficiency than the traditional technique as well as avoiding UV irradiation of the cytoplast. 650 $aoocytes 653 $aBovine 653 $aCytoplasts 700 1 $aOLIVEIRA, C. S. 700 1 $aTETZNER, T. A. D. 700 1 $aSOUZA, M. M. 700 1 $aLIMA, M. R. de 700 1 $aNICIURA, S. C. M. 700 1 $aGARCIA, J. M.
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