5 research outputs found

    Rapid cell type-specific nascent proteome labeling in Drosophila

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    Controlled protein synthesis is required to regulate gene expression and is often carried out in a cell type-specific manner. Protein synthesis is commonly measured by labeling the nascent proteome with amino acid analogs or isotope-containing amino acids. These methods have been difficult to implement in vivo as they require lengthy amino acid replacement procedures. O-propargyl-puromycin (OPP) is a puromycin analog that incorporates into nascent polypeptide chains. Through its terminal alkyne, OPP can be conjugated to a fluorophore-azide for directly visualizing nascent protein synthesis, or to a biotin-azide for capture and identification of newly-synthesized proteins. To achieve cell type-specific OPP incorporation, we developed phenylacetyl-OPP (PhAc-OPP), a puromycin analog harboring an enzyme-labile blocking group that can be removed by penicillin G acylase (PGA). Here, we show that cell type-specific PGA expression in Drosophila can be used to achieve OPP labeling of newly-synthesized proteins in targeted cell populations within the brain. Following a brief 2 hr incubation of intact brains with PhAc-OPP, we observe robust imaging and affinity purification of OPP-labeled nascent proteins in PGA-targeted cell populations. We apply this method to show a pronounced age-related decline in neuronal protein synthesis in the fly brain, demonstrating the capability of PhAc-OPP to quantitatively capture in vivo protein synthesis states. This method, which we call POPPi (PGA-dependent OPP incorporation), should be applicable for rapidly visualizing protein synthesis and identifying nascent proteins synthesized under diverse physiological and pathological conditions with cellular specificity in vivo

    Socioeconomic, Clinical, and Molecular Features of Breast Cancer Influence Overall Survival of Latin American Women

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    Molecular profile of breast cancer in Latin-American women was studied in five countries: Argentina, Brazil, Chile, Mexico, and Uruguay. Data about socioeconomic characteristics, risk factors, prognostic factors, and molecular subtypes were described, and the 60-month overall cumulative survival probabilities (OS) were estimated. From 2011 to 2013, 1,300 eligible Latin-American women 18 years or older, with a diagnosis of breast cancer in clinical stage II or III, and performance status ≊̞1 were invited to participate in a prospective cohort study. Face-to-face interviews were conducted, and clinical and outcome data, including death, were extracted from medical records. Unadjusted associations were evaluated by Chi-squared and Fisher’s exact tests and the OS by Kaplan–Meier method. Log-rank test was used to determine differences between cumulative probability curves. Multivariable adjustment was carried out by entering potential confounders in the Cox regression model. The OS at 60 months was 83.9%. Multivariable-adjusted death hazard differences were found for women living in Argentina (2.27), Chile (1.95), and Uruguay (2.42) compared with Mexican women, for older (≄60 years) (1.84) compared with younger (≀40 years) women, for basal-like subtype (5.8), luminal B (2.43), and HER2-enriched (2.52) compared with luminal A subtype, and for tumor clinical stages IIB (1.91), IIIA (3.54), and IIIB (3.94) compared with stage IIA women. OS was associated with country of residence, PAM50 intrinsic subtype, age, and tumor stage at diagnosis. While the latter is known to be influenced by access to care, including cancer screening, timely diagnosis and treatment, including access to more effective treatment protocols, it may also influence epigenetic changes that, potentially, impact molecular subtypes. Data derived from heretofore understudied populations with unique geographic ancestry and sociocultural experiences are critical to furthering our understanding of this complexity. Copyright © 2022 de Almeida, CortĂ©s, Vilensky, Valenzuela, Cortes-Sanabria, de Souza, Barbeito, Abdelhay, Artagaveytia, Daneri-Navarro, Llera, MĂŒller, Podhajcer, Velazquez, Alcoba, Alonso, Bravo, Camejo, Carraro, Castro, Cataldi, Cayota, Cerda, Colombo, Crocamo, Del Toro-Arreola, Delgadillo-Cristerna, Delgado, Breitenbach, FernĂĄndez, FernĂĄndez, FernĂĄndez, Franco-Topete, Gaete, GĂłmez, Gonzalez-Ramirez, Guerrero, Gutierrez-Rubio, Jalfin, Lopez-Vazquez, Loria, MĂ­guez, Moran-Mendoza, Morgan-Villela, Mussetti, Nagai, Oceguera-Villanueva, Reis, Retamales, Rodriguez, Rosales, Salas-Gonzalez, Segovia, Sendoya, Silva-Garcia, Viña, Zagame, Jones, Szklo and United States-Latin American Cancer Research Network (US-LACRN).Fil: de Almeida, Liz Maria. Instituto Nacional de CĂĄncer; BrasilFil: CortĂ©s, Sandra. Pontificia Universidad CatĂłlica de Chile; ChileFil: Vilensky, Marta. Instituto de OncologĂ­a Angel Roffo; ArgentinaFil: Valenzuela, Olivia. Universidad de Sonora; MĂ©xicoFil: Cortes-Sanabria, Laura. Hospital de Especialidades, CMNO-IMSS; MĂ©xicoFil: de Souza, Mirian. Instituto Nacional de CĂĄncer; BrasilFil: Barbeito, Rafael Alonso. Facultad de Medicina; ArgentinaFil: Abdelhay, Eliana. Instituto Nacional de CĂĄncer; BrasilFil: Artagaveytia, Nora. Hospital de ClĂ­nicas Manuel Quintela. Universidad de la RepĂșblica; UruguayFil: Daneri-Navarro, Adrian. Universidad de Guadalajara; MĂ©xicoFil: Llera, Andrea S. CONICET. FundaciĂłn Instituto Leloir; ArgentinaFil: MĂŒller, Bettina. Instituto Nacional del CĂĄncer; ArgentinaFil: Podhajcer, Osvaldo L. CONICET. FundaciĂłn Instituto Leloir; ArgentinaFil: Velazquez, Carlos. Universidad de Sonora; MĂ©xicoFil: Alcoba, Elsa. Hospital Municipal de OncologĂ­a MarĂ­a Curie; ArgentinaFil: Alonso, Isabel. Centro Hospitalario Pereira Rossell; ArgentinaFil: Bravo, Alicia I. Hospital Regional de Agudos Eva PerĂłn; ArgentinaFil: Camejo, Natalia. Hospital de ClĂ­nicas Manuel Quintela. Universidad de la RepĂșblica; UruguayFil: Carraro, Dirce Maria. AC Camargo Cancer Center; BrasilFil: Castro, MĂłnica. Instituto de OncologĂ­a Angel Roffo; ArgentinaFil: Cataldi, Sandra. Instituto Nacional de CĂĄncer; UruguayFil: Cayota, Alfonso. Institut Pasteur de Montevideo; UruguayFil: Cerda, Mauricio. Universidad de Chile; ChileFil: Colombo, Alicia. Universidad de Chile; ChileFil: Crocamo, Susanne. Instituto Nacional de CĂĄncer; BrasilFil: Del Toro-Arreola, Alicia. Universidad de Guadalajara; MĂ©xicoFil: Delgadillo-Cristerna, Raul. Hospital de Especialidades. CMNO-IMSS; MĂ©xicoFil: Delgado, Lucia. Hospital de ClĂ­nicas Manuel Quintela; UruguayFil: Breitenbach, Marisa Dreyer. Universidade do Estado do Rio de Janeiro; BrasilFil: FernĂĄndez, Elmer. Universidad CatĂłlica de CĂłrdoba. CONICET. Centro de Investigaciones en BioquĂ­mica ClĂ­nica e Inmunologia; ArgentinaFil: FernĂĄndez, Jorge. Instituto de Salud PĂșblica; ChileFil: FernĂĄndez, Wanda. Hospital San Borja ArriarĂĄn; ChileFil: Franco-Topete, Ramon A. OPD Hospital Civil de Guadalajara. Universidad de Guadalajara; MĂ©xicoFil: Gaete, Fancy. Hospital Luis Tisne; ChileFil: GĂłmez, Jorge. Texas A&M University; Estados UnidosFil: Gonzalez-Ramirez, Leivy P. Universidad de Guadalajara; MĂ©xicoFil: Guerrero, Marisol. Hospital San JosĂ©; ChileFil: Gutierrez-Rubio, Susan A. Universidad de Guadalajara; MĂ©xicoFil: Jalfin, Beatriz. Hospital Regional de Agudos Eva PerĂłn; ArgentinaFil: Lopez-Vazquez, Alejandra. Universidad de Sonora; MĂ©xicoFil: Loria, Dora. Instituto de OncologĂ­a Angel Roffo; ArgentinaFil: MĂ­guez, Silvia. Hospital Municipal de OncologĂ­a MarĂ­a Curie; ArgentinaFil: Moran-Mendoza, Andres de J. Hospital de Gineco-Obstetricia CMNO-IMSS; MĂ©xicoFil: Morgan-Villela, Gilberto. Hospital de Especialidades. CMNO-IMSS; MĂ©xicoFil: Mussetti, Carina. Registro Nacional de Cancer; UruguayFil: Nagai, Maria Aparecida. Instituto de CĂąncer de SĂŁo Paulo; BrasilFil: Oceguera-Villanueva, Antonio. Instituto Jalisciense de Cancerologia; MĂ©xicoFil: Reis, Rui M. Hospital de CĂąncer de Barretos; BrasilFil: Retamales, Javier. Grupo OncolĂłgico Cooperativo Chileno de InvestigaciĂłn; ChileFil: Rodriguez, Robinson. Hospital Central de las Fuerzas Armadas; UruguayFil: Rosales, Cristina, Hospital Municipal de OncologĂ­a MarĂ­a Curie; ArgentinaFil: Salas-Gonzalez, Efrain. Hospital San JosĂ©; ChileFil: Segovia, Laura. Hospital Barros Luco Trudeau; ChileFil: Sendoya, Juan M. CONICET. FundaciĂłn Instituto Leloir,; ArgentinaFil: Silva-Garcia, Aida A. OPD Hospital Civil de Guadalajara. Universidad de Guadalajara; MĂ©xicoFil: Viña, Stella. Instituto de OncologĂ­a Angel Roffo; ArgentinaFil: Zagame, Livia. Instituto Jalisciense de Cancerologia; MĂ©xicoFil: Jones, Beth. Yale University. Yale School of Public Health; Estados UnidosFil: Szklo, MoysĂ©s. Johns Hopkins University. Johns Hopkins Bloomberg School of Public Health; Estados Unido
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