Introducción: El cáncer de mama (CM) es la segunda neoplasia en Perú (19.5%). Se evaluaron los porcentajes de poblaciones inmunocompetentes y el perfil Th1/Th2/Th17 en sangre periférica (SP) y microambiente tumoral (MT) de pacientes peruanas en diferentes estadios de CM diagnosticadas recientemente y sin tratamiento. Metodología: Se utilizaron muestras de SP y biopsias, obtenidas en el Instituto Nacional de Enfermedades Neoplásicas (2018), diagnosticadas con CM en estadios II (n=6), III (n=8) y IV (n=4). Obtuvimos células mononucleares de SP (PBMC) y células mononucleares del MT (MTMC). En ambos casos, las células se cultivaron por 24h y luego se realizó citometría de flujo para macrófagos M1 (CD68+), M2 (CD163+), células T (CD4+/CD8+) y células dendríticas mieloides (DCm, CD11c+) y plasmocitoides (DCp, CD11c-). Además, se obtuvieron sobrenadantes de cultivo para medir IF...

Recent publications have shown that active RNA polymerase (RNAP) from Mycobacterium tuberculosis (MtbRNAP) can be produced by expressing all four subunits in a single recombinant Escherichia coli strain [1–3]. By reducing the number of plasmids and changing the codon usage of the Mtb genes in the co-expression system published by Banerjee et al. [1], we present a simplified, detailed and reproducible protocol for the purification of recombinant MtbRNAP containing the ω subunit. Moreover, we describe the formation of ternary elongation complexes (TECs) with a short fluorescence-labeled RNA primer and DNA oligonucleotides, suitable for transcription elongation studies.

This work was supported by Fondecyt 11110534 (to M.B.), 1151274 (to M.B.), 11130263 (to C.A.M.W.), Anillo 1107 (to M.B.), and Chile and Fondecyt 196–2013 (to D.G.G.) CONCYTEC, Perú. This research was partially supported by the supercomputing infrastructure of the NLHPC (ECM-02) at Universidad de Chile. A.B. was supported by Conicyt master fellowship 22121199. We gratefully acknowledge Professor Alexander Vologodskii from New York University for helpful comments and advice. Travel for C.B. to and from Chile and Peru was also partially supported by the Howard Hughes Medical Institute, NIH grant R01GM032543, and the U.S. Department of Energy Office of Basic Energy Sciences Nanomachine Program under contract no. DE-AC02-05CH11231.