IL-12 Can Target Human Lung Adenocarcinoma Cells and Normal Bronchial Epithelial Cells Surrounding Tumor Lesions

BACKGROUND: Non small cell lung cancer (NSCLC) is a leading cause of cancer death. We have shown previously that IL-12rb2 KO mice develop spontaneously lung adenocarcinomas or bronchioalveolar carcinomas. Aim of the study was to investigate i) IL-12Rbeta2 expression in human primary lung adenocarcinomas and in their counterparts, i.e. normal bronchial epithelial cells (NBEC), ii) the direct anti-tumor activity of IL-12 on lung adenocarcinoma cells in vitro and vivo, and the mechanisms involved, and iii) IL-12 activity on NBEC. METHODOLOGY/PRINCIPAL FINDINGS: Stage I lung adenocarcinomas showed significantly (P = 0.012) higher frequency of IL-12Rbeta2 expressing samples than stage II/III tumors. IL-12 treatment of IL-12R(+) neoplastic cells isolated from primary adenocarcinoma (n = 6) inhibited angiogenesis in vitro through down-regulation of different pro-angiogenic genes (e.g. IL-6, VEGF-C, VEGF-D, and laminin-5), as assessed by chorioallantoic membrane (CAM) assay and PCR array. In order to perform in vivo studies, the Calu6 NSCLC cell line was transfected with the IL-12RB2 containing plasmid (Calu6/beta2). Similar to that observed in primary tumors, IL-12 treatment of Calu6/beta2(+) cells inhibited angiogenesis in vitro. Tumors formed by Calu6/beta2 cells in SCID/NOD mice, inoculated subcutaneously or orthotopically, were significantly smaller following IL-12 vs PBS treatment due to inhibition of angiogenesis, and of IL-6 and VEGF-C production. Explanted tumors were studied by histology, immuno-histochemistry and PCR array. NBEC cells were isolated and cultured from lung specimens of non neoplastic origin. NBEC expressed IL-12R and released constitutively tumor promoting cytokines (e.g. IL-6 and CCL2). Treatment of NBEC with IL-12 down-regulated production of these cytokines. CONCLUSIONS: This study demonstrates that IL-12 inhibits directly the growth of human lung adenocarcinoma and targets the adjacent NBEC. These novel anti-tumor activities of IL-12 add to the well known immune-modulatory properties of the cytokine and may provide a rational basis for the development of a clinical trial

Realidad nacional de la producción y consumo de hortalizas 2020, en tiempos de la COVID-19

En el actual contexto de complejidad socioeconómica y sanitaria, con el objetivo de disponer de información actualizada respecto de la realidad nacional de la producción y consumo de hortalizas, de las preferencias y hábitos de consumo de vegetales de la población argentina, así como también de los cambios producidos en la alimentación, por la pandemia global, COVID-19, se realizó una encuesta, a nivel nacional, titulada: "Tendencias del consumo de hortalizas en Argentina, en tiempos del Coronavirus", que fue respondida por 1694 encuestados. La población argentina cuenta con una gran predisposición a la realización de huertas urbanas familiares ante el nuevo contexto, la misma tiene preferencias de consumo claras, valora los beneficios para la salud del consumo de hortalizas y considera que cuenta con una proporción adecuada de vegetales en su dieta. No obstante, surge de la encuesta, que el consumo real es muy inferior a la ingesta recomendada por parte de entidades nacionales e internacionales. Por tal motivo, existe una importante asimetría, entre lo que la población cree que consume y lo que realmente ingiere Este hecho conlleva a intensificar acciones, por parte de diversos organismos e instituciones, tendientes a lograr mayor conciencia de dichas diferencias y promover el consumo de hortalizas en cantidad, calidad y diversidad, que mejore el bienestar, reduzca el riesgo de desarrollo de enfermedades y beneficie la calidad de vida de la población.In the present complex socioeconomical and sanitary context, with the aim of having updated information on the national reality of vegetable production and consumption, of the Argentinian population’s preferences and consumption habits of vegetables as well as the changes produced in their diet due to the global COVID-19 pandemics, a national survey, titled “Trends in the consumption of vegetables in Argentina in Coronavirus times” was carried out obtaining 1694 responses. The Argentinian population has had a great inclination to start growing, urban family gardens in this new context; it has clear consumption preferences, it values the benefits of consuming vegetables for health reasons and considers it has an adequate proportion of vegetables in its diet. However, the survey shows that real consumption is much lower than that recommended by national and international entities arises from the survey. Therefore, there is a great asymmetry between what the population believes it consumes and what it actually eats. This fact implies intensifying actions towards raising greater awareness of these differences and promoting adequate consumption of quantity, quality and diversity of vegetables to improve well-being, reduce the risk of developing illnesses and benefit the population’s life quality by various organizations and institutions.Fil: Castagnino, Ana Maria. Universidad Nacional del Centro de la Provincia de Buenos Aires; ArgentinaFil: Diaz, Karina Elizabeth. Universidad Nacional del Centro de la Provincia de Buenos Aires; ArgentinaFil: Bazán, P.. Universidad Nacional de San Luis; ArgentinaFil: Castro Luna, Adolfo Eduardo. Universidad Nacional de San Luis; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Martinoia, Gabriela Ivone. Universidad Nacional del Centro de la Provincia de Buenos Aires; ArgentinaFil: Marina, Javier Alejandro. Universidad Nacional del Centro de la Provincia de Buenos Aires; ArgentinaFil: Echeverría, S.. Universidad Nacional del Centro de la Provincia de Buenos Aires; ArgentinaFil: Galizio, Rodrigo. Universidad Nacional del Centro de la Provincia de Buenos Aires; ArgentinaFil: Rosini, M. B.. Universidad Nacional del Centro de la Provincia de Buenos Aires; ArgentinaFil: Rogers, William John. Universidad Nacional del Centro de la Provincia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Rubel, Irene Albertina. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ingeniería Olavarría. Grupo Tecnologías de Semillas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Benson, Silvia. Universidad Nacional del Centro de la Provincia de Buenos Aires; ArgentinaFil: Díaz, H.. Universidad Nacional del Centro de la Provincia de Buenos Aires; ArgentinaFil: Reina, R.. Universidad Nacional del Centro de la Provincia de Buenos Aires; Argentin

Cyclohexenylphenyldiazene: A Simple Surrogate of the Azobenzene Photochromic Unit

We have carried out an experimental and computational study on the ground- and excited-state photochemical and photophysical properties of (1-cyclohexenyl)phenyldiazene (CPD), a species formally derived from azobenzene in which one of the phenyl rings is replaced by a 1-cyclohexene substituent. The results show that CPD does substantially behave like azobenzene, but with a higher (~70%) ZE (n*) photoisomerization quantum yield, calling for CPD as an effective alternative of azobenzene itself with new functionalization possibilities. By use of state-of-the-art ab initio CASPT2//CASSCF minimum energy path computations, we have identified the most efficient decay and isomerization routes of the absorbing singlet (*), S1 (n*), T1, and S0 states of CPD. The resulting mechanistic scheme agrees with experimental findings and provides a rationale of the observed photoisomerization quantum yields. Furthermore, this study provides a deep insight on the photophysical and photochemical properties of compounds based on the -N=N- double bond which supplies a general model for the photoreactivity of azobenzene-type compounds in general. This is expected to be a useful guideline for the design of novel photoreactive azo compounds

New nature-inspired hybrids targeting the Nrf2-HO1 pathway protect retinal pigment epithelial cells under stress conditions.

Purpose: An impairment in the Nuclear factor E2-related factor 2 (Nrf2) pathway, a master regulator of stress response in retinal pigment epithelium (RPE), is suggested to contribute to the Age-related Macular Degeneration (AMD) pathogenesis; Nrf2-activators may be of interest in AMD and other pathologies featured by oxidative stress. We tested in vitro in ARPE19 cells some novel Nature-Inspired Hybrids (NIHs) for their capability to activate Nrf2 pathway and to provide protection from AMD-related stressors. Methods: Four NIHs (5μM) were individually tested for their pharmacological activity and SAR in ARPE-19 cells. Control cells (CTR) were treated with the solvent DMSO. The Nrf2 activator dimethyl-fumarate (DMF; 10μM) was used as a positive control. Nrf2 nuclear translocation was evaluated by Western blotting. Among Nrf2-target genes, the Heme-Oxygenase 1 (HO-1) expression was measured by real-time qPCR and Western blotting. The NIHs’ effects on ROS levels and cell viability were evaluated following either a short term exposure to H2O2 (0.3mM), or a long term exposure to MG132 (5μM) + Bafilomycin (50nM) (M+B, autophagy inhibitor) by fluorimetric assays. We performed ≥3 independent replicates for each experiment. Prism was used for statistical analysis (one-way or two-way ANOVA followed by Dunnett’s multiple comparisons test). Results: ARPE-19 cells well tolerate all NIHs (no cell viability changes after 48 hrs treatments; n=6). NIHs activate Nrf2 pathway with different times and intensities among molecules. The most promising NIH, lead compound (NIH1) provides: i) Nrf2 nuclear translocation after 3 hrs (+65.8±12.4% vs CTR), accompanied by a 4.5-fold increase of HO-1 mRNA total level with respect to control cells (n=3, p<0.05); ii) a long-lasting (up to 48 hrs) increase of HO-1 protein levels (+311.3±30% vs CTR; n=3, p<0.01); iii) a direct antioxidant effect against 4 hrs H2O2 treatment (n=4, p<0.0001); iv) an increase of cell viability under 48 hrs MG132+Bafilomycin (M+B) stress (CTR: 100±0.6%; M+B: 75.6±0.94%; NIH1+M+B: 90.1±1.35%; n=4-8, p<0.0001). Conclusion: A positive modulation of Nrf2-pathway by NIHs is protective in RPE cells, encouraging further studies on their potential use in pathologies featured by oxidative stress. Layman Abstract (optional) Retina is subjected to stressors (such as free radicals) for the entire life span, and endogenous protective factors are essential to preserve this tissue. In particular, the Nuclear factor E2-related factor 2 (Nrf2) pathway is a master regulator of the stress response in the retina, and especially in the retinal pigment epithelium (RPE). Nrf2 pathway is impaired in aging and in some ocular pathologies, such as Age-related Macular Degeneration (AMD), and molecules able to stimulate Nrf2 are interesting as new potential drugs. We studied some new nature-inspired hybrids that activate Nrf2 pathway, and we found they can protect RPE cells in vitro from AMD-related stressors. Our findings encourage further studies on the potential use of these molecules in AMD and other pathologies featured by oxidative stress

New nature-inspired hybrids activating the Nrf2-HO1 pathway in retinal pigment epithelial cells and their potential use in pathologies featured by oxidative stress

Purpose: Among the earliest factors triggering Age-related Macular Degeneration (AMD) is the degeneration of retinal pigment epithelium (RPE). A primary system used by RPE to neutralize oxidative stress and maintain cellular homeostasis is the nuclear factor E2-related factor 2 (Nrf2) pathway. We aimed to further test some novel Nature-Inspired Hybrids (NIH) endowed with anti-oxidant properties, in order to evaluate their capability to activate Nrf2-pathway and to promote protection in RPE cells. Methods: ARPE-19 cells were exposed to NIH (5μM) for increasing times (from 3 to 48 hours). Dimethyl-fumarate (DMF; 10μM), a well-known Nrf2 activator, was used as a positive control. The Nrf2-pathway activation was evaluated by studying Nrf2 protein nuclear translocation and Heme-Oxygenase 1 (HO-1) expression (mRNA and protein, with real time PCR and Western blotting, respectively). Cell viability of ARPE-19 exposed to the NIH in the presence/absence of AMD-related stressors was evaluated by various techniques (MTT, LDH and PrestoBlue® assay). Results: The NIH are well tolerated by ARPE-19 cells. The NIH presenting in their structure the chemical active group(s) responsible for the Nrf2-pathway activation (catechol group and/or Michael acceptor) induce Nrf2 nuclear translocation, suggesting an interference in Nrf2 protein degradation process. The same NIH upregulate HO-1 expression, although with levels and/or time-courses that vary among molecules, with potential outcomes on the cellular stress response. Conclusion: A positive modulation of Nrf2-pathway by NIH may be protective in RPE cells, encouraging further studies on their potential use in pathologies featured by oxidative stress