👤 Javier C Angulo

Indole-3-carbinol (I3C) is a metabolic derivative of glucobrassicin found in cruciferous vegetables. Known for its anticarcinogenic properties, I3C has been shown to target the NEDD4 family HECT E3 ligases, NEDD4-1 and WWP1, yet Show less

Protein kinase D (PKD) family members play controversial roles in prostate cancer (PC). Thus, PKD1 is nearly absent in advanced tumours, where PKD2 and PKD3 are upregulated. Additionally, consequences of activation of these kinases on PC progression remain largely unclear. Here, we first investigated PKD function on PC cell motility, analysing the underlying molecular mechanisms. We find a striking decrease of Snail levels after PKD inhibition followed by cell migration and invasion impairment, demonstrating an unprecedented role of PKD activity on the regulation of this key transcription factor in PC progression. Specifically, we show that PKD2 activity mediates the effects of MEK/ERK pathway on Snail expression, establishing a joint function of ERK/PKD2/Snail cascade in PC cell invasion regulation. These results led us to address the clinical relevance of the correlation between PKD2 and ERK activities with Snail abundance in samples from PC patients at different stages, analysing its impact on tumour prognosis and patients´ survival. Importantly, this is the first study defining a direct correlation between active PKD2 and Snail levels, further linked to ERK activity. We also evidence that PKD2 activity is associated with important poor prognostic factors. Thus, PC patients with the expression pattern: active PKD2 Show less

We focus on this study in designing an alternative technique for obtaining mesenchymal stem cells (MSCs) from residual tissue, Hoffa fat, in arthroscopic procedures. Two males and two females were included, and underwent knee arthroscopy; a sample of infrapatellar adipose tissue was obtained with basket forceps. The primary culture was made using the explant method and the culture media: DMEM-high glucose, supplemented with 10% of inactivated human allogeneic serum. All the cellular cultures remained under culture conditions for three weeks, after that by flow cytometry the cells were characterized by MSCs antibody panel: CD105, CD73 and CD90. Subsequently, in the first pass, the MSCs were cultured in commercial human chondrogenic, osteogenic and adipogenic mediums, respectively. After primary culture, we obtained on average 95,600.00 ± 7,233.26 cells/cm2, and the duplication time of MSCs isolate from Hoffa fat pad was established in 39 hours. By flow cytometry, we found that surface markers percentage for expanded MSCs (CD105, CD73, CD90) in primary culture significantly increased and its morphology was fibroblastic-like. After differentiation culture which was made in the first pass, by immunofluorescence, we obtained positive cell markers for three lineages of differentiation, adipocytes: LPL protein, osteocytes: RUNX2, Osteopontin, chondrocytes: SOX9, Aggrecan and COL2A1. We managed to isolate a significant number of MSCs from this source using an easy method to implement and minimal nutrient supplementation, with high potential for differentiation to mature mesenchymal tissues and potential use in basic experimental, preclinical and even clinical research. Show less

We have screened small molecule libraries specifically for inhibitors that target WWP2, an E3 ubiquitin ligase associated with tumour outgrowth and spread. Selected hits demonstrated dose-dependent WWP2 inhibition, low micromolar IC50 values, and inhibition of PTEN substrate-specific ubiquitination. Binding to WWP2 was confirmed by ligand-based NMR spectroscopy. Furthermore, we used a combination of STD NMR, the recently developed DEEP-STD NMR approach, and docking calculations, to propose for the first time an NMR-validated 3D molecular model of a WWP2-inhibitor complex. These first generation WWP2 inhibitors provide a molecular framework for informing organic synthetic approaches to improve activity and selectivity. Show less

The effect of a 10-week supplementation with polyunsaturated fatty acids [via sunflower oil/DHA-rich algae (SUNA) or linseed oil/DHA-rich algae (LINA) enriched diets] versus saturated fatty acids (SAT) of lactating German Holstein dairy cows in mid-lactation on expression patterns of lipid metabolism-associated genes and gene products in hepatic, longissimus muscle and subcutaneous/perirenal/omental adipose tissue was assessed. Most pronounced transcriptomic responses to dietary PUFA were obtained in hepatic [down-regulated ACACA (FC = 0.83, SUNA; FC = 0.86, LINA), FADS1 (FC = 0.60, SUNA; FC = 0.72, LINA), FADS2 (FC = 0.64, SUNA; FC = 0.79, LINA), FASN (FC = 0.64, SUNA; FC = 0.72, LINA), SCD (FC = 0.37, SUNA; FC = 0.47, LINA) and SREBF1 (FC = 0.79, SUNA, LINA) expression] and omental adipose [up-regulated ACACA (FC = 1.58, SUNA; FC = 1.22, LINA), ADFP (FC = 1.33, SUNA; FC = 1.32, LINA), CEBPA (FC = 1.75, SUNA; FC = 1.40, LINA), FASN (FC = 1.57, SUNA; FC = 1.21, LINA), LPL (FC = 1.50, SUNA; FC = 1.20, LINA), PPARG (FC = 1.36, SUNA; FC = 1.12, LINA), SCD (FC = 1.41, SUNA; FC = 1.17, LINA) and SREBF1 (FC = 1.56, SUNA; FC = 1.18, LINA) expression] tissue. Interestingly, gene/gene product associations were comparatively low in hepatic and omental adipose tissue compared with longissimus muscle, perirenal adipose and subcutaneous adipose tissue, indicating matches only in regard to minor concentrations of SCD product 18:1c9, FADS1 product 20:4n-6 and FADS2 product 18:3n-6 in hepatic tissue, and higher concentrations of ACACA and FASN gene products 12:0 and 14:0 and SCD product 18:2c9,t11 in omental adipose tissue. Whereas all analyzed tissues accumulated dietary PUFA and their ruminally generated biohydrogenation products, tissue-divergent preferences for certain fatty acids were identified. This descriptive study reports tissue-divergent effects of dietary PUFA and outlines the significance of a PUFA intervention with regard to dairy cows' nutritional management. Show less