Sequential Colocalization of ERa, PR, and AR Hormone Receptors Using Confocal Microscopy Enables New Insights into Normal Breast and Prostate Tissue and Cancers

Multiplex immunohistochemistry (mIHC) use markers staining different cell populations applying widefield optical microscopy. Resolution is low not resolving subcellular co-localization. We sought to colocalize markers at subcellular level with antibodies validated for clinical diagnosis, including the single secondary antibody (combination of anti-rabbit/mouse-antibodies) used for diagnostic IHC with any primary antibody, and confocal microscopy. We explore colocalization in the nucleus (ColNu) of nuclear hormone receptors (ERa, PR, and AR) along with the baseline marker p63 in paired samples of breast and prostate tissues. We established ColNu mIHCF as a reliable technique easily implemented in a hospital setting. In ERa+ breast cancer, we identified different colocalization patterns (nuclear or cytoplasmatic) with PR and AR on the luminal epithelium. A triple-negative breast-cancer case expressed membrane-only ERa. A PR-only case was double positive PR/p63. In normal prostate, we identified an ERa+/p63+/AR-negative distinct population. All prostate cancer cases characteristically expressed ERa on the apical membrane of the AR+ epithelium. We confirmed this using ERa IHC and needle-core biopsies. ColNu mIHCF is feasible and already revealed a new marker for prostate cancer and identified sub-patterns in breast cancer. It could be useful for pathology as well as for functional studies in normal prostate and breast tissuesThis study has been supported by projects from the State Research Agency (AEI), and Ministry of Science and Innovation, Spain, MINECO-FEDER PID2019-110437RB-I00 (to CVA) and Instituto de Salud Carlos III (ISCIII)-FEDER PI19/01316 (to JMCT). Financial support code: ED431G 2019/02 from the Xunta de Galicia and the European Union (European Regional Development Fund - ERDF) to the Centro singular de Investigación de Galicia accreditation 2019-2022 is gratefully acknowledgedS

Estimación in vivo de la canal porcina por el método de ultrasonografía: Un Enfoque de la Ecointensificación en Bioeconomia Porcina

En este estudio se analiza la relación entre el espesor de grasa subcutánea (EGS), espesor muscular (EM) y espesor de grasa subcutánea y espesor muscular juntos (EGSM), medidos con ultrasonido en un solo punto anatómico de cerdos vivos y los parámetros más importantes de la canal . En nuestro experimento utilizamos cerdos híbridos  de las  razas Pietrain, Landrace y Yorkshire en cantidad de 25 animales de ambos sexos.   Los resultados del estudio  sugieren que los parámetros de la composición de la canal aquí analizados están altamente correlacionados con el espesor de la grasa subcutánea medida con el  ultrasonido en la última costilla del musculo longuísimo dorsal en el lado izquierdo del cerdo vivo (r = 0.53 - 0.67). La correlación múltiple del modelo de predicción de regresión lineal entre los parámetros de la canal y el espesor de grasa dorsal fue (0.74) y el coeficiente de determinación mostró suficiente capacidad de predicción (R2 = 0.55). Los modelos de predicción de regresión lineal de los componentes de la canal con el espesor muscular obtuvieron baja capacidad de predicción (R2 = 0.23). Igualmente en el modelo de regresión del espesor de grasa subcutánea y espesor muscular juntos ( R2= 0.25).  Entre las correlaciones más importantes encontradas están: lomo derecho, carne clase A (r = 0.80), peso vivo y canal entera (r = 0.85) paleta derecha y carne clase B (r = 0.88) siendo este patrón repetitivo en todas las correlaciones de las mediciones ultrasonografica y los componentes de la canal

Inhibition of iNOS as a Novel Effective Targeted Therapy Against Triple-Negative Breast Cancer

INTRODUCTION: Triple-negative breast cancer (TNBC) is an aggressive form of breast cancer with no effective targeted therapy. Inducible nitric oxide synthase (iNOS) is associated with poor survival in patients with breast cancer by increasing tumor aggressiveness. This work aimed to investigate the potential of iNOS inhibitors as a targeted therapy for TNBC. We hypothesized that inhibition of endogenous iNOS would decrease TNBC aggressiveness by reducing tumor initiation and metastasis through modulation of epithelial-mesenchymal transition (EMT)-inducing factors. METHODS: iNOS protein levels were determined in 83 human TNBC tissues and correlated with clinical outcome. Proliferation, mammosphere-forming efficiency, migration, and EMT transcription factors were assessed in vitro after iNOS inhibition. Endogenous iNOS targeting was evaluated as a potential therapy in TNBC mouse models. RESULTS: High endogenous iNOS expression was associated with worse prognosis in patients with TNBC by gene expression as well as immunohistochemical analysis. Selective iNOS (1400 W) and pan-NOS (L-NMMA and L-NAME) inhibitors diminished cell proliferation, cancer stem cell self-renewal, and cell migration in vitro, together with inhibition of EMT transcription factors (Snail, Slug, Twist1, and Zeb1). Impairment of hypoxia-inducible factor 1α, endoplasmic reticulum stress (IRE1α/XBP1), and the crosstalk between activating transcription factor 3/activating transcription factor 4 and transforming growth factor β was observed. iNOS inhibition significantly reduced tumor growth, the number of lung metastases, tumor initiation, and self-renewal. CONCLUSIONS: Considering the effectiveness of L-NMMA in decreasing tumor growth and enhancing survival rate in TNBC, we propose a targeted therapeutic clinical trial by re-purposing the pan-NOS inhibitor L-NMMA, which has been extensively investigated for cardiogenic shock as an anti-cancer therapeutic

Multi-stage delivery nano-particle systems for therapeutic applications

BACKGROUND: The daunting task for drug molecules to reach pathological lesions has fueled rapid advances in Nanomedicine. The progressive evolution of nanovectors has led to the development of multi-stage delivery systems aimed at overcoming the numerous obstacles encountered by nanovectors on their journey to the target site. SCOPE OF REVIEW: This review summarizes major findings with respect to silicon-based drug delivery vectors for cancer therapeutics and imaging. Based on rational design, well established silicon technologies have been adapted for the fabrication of nanovectors with specific shapes, sizes, and porosities. These vectors are part of a multi-stage delivery system that contains multiple nano-components, each designed to achieve a specific task with the common goal of site-directed delivery of therapeutics. MAJOR CONCLUSIONS: Quasi-hemispherical and discoidal silicon microparticles are superior to spherical particles with respect to margination in the blood, with particles of different shapes and sizes having unique distributions in vivo. Cellular adhesion and internalization of silicon microparticles is influenced by microparticle shape and surface charge, with the latter dictating binding of serum opsonins. Based on in vitro cell studies, the internalization of porous silicon microparticles by endothelial cells and macrophages is compatible with cellular morphology, intracellular trafficking, mitosis, cell cycle progression, cytokine release, and cell viability. In vivo studies support superior therapeutic efficacy of liposomal encapsulated siRNA when delivered in multi-stage systems compared to free nanoparticles

Estimación in vivo de la canal porcina por el método de ultrasonografía: Un Enfoque de la Ecointensificación en Bioeconomia Porcina

Revista Iberoamericana De Bioeconomía Y Cambio Climático, 5(10), 1278–1294En este estudio se analiza la relación entre el espesor de grasa subcutánea (EGS), espesor muscular (EM) y espesor de grasa subcutánea y espesor muscular juntos (EGSM), medidos con ultrasonido en un solo punto anatómico de cerdos vivos y los parámetros más importantes de la canal . En nuestro experimento utilizamos cerdos híbridos de las razas Pietrain, Landrace y Yorkshire en cantidad de 25 animales de ambos sexos. Los resultados del estudio sugieren que los parámetros de la composición de la canal aquí analizados están altamente correlacionados con el espesor de la grasa subcutánea medida con el ultrasonido en la última costilla del musculo longuísimo dorsal en el lado izquierdo del cerdo vivo (r = 0.53 - 0.67). La correlación múltiple del modelo de predicción de regresión lineal entre los parámetros de la canal y el espesor de grasa dorsal fue (0.74) y el coeficiente de determinación mostró suficiente capacidad de predicción (R2 = 0.55). Los modelos de predicción de regresión lineal de los componentes de la canal con el espesor muscular obtuvieron baja capacidad de predicción (R2 = 0.23). Igualmente en el modelo de regresión del espesor de grasa subcutánea y espesor muscular juntos ( R2= 0.25). Entre las correlaciones más importantes encontradas están: lomo derecho, carne clase A (r = 0.80), peso vivo y canal entera (r = 0.85) paleta derecha y carne clase B (r = 0.88) siendo este patrón repetitivo en todas las correlaciones de las mediciones ultrasonografica y los componentes de la canal.Universidad Nacional Autónoma de Nicaragua, León. Escuela de Ciencias Agrarias y Veterinarias. Departamento de Agroecologia