Underutilized Crocus Sativus L. Flowers: A Hidden Source of Sustainable High Value-Added Ingredients.

Crocus sativus L. is used as a spice due to its organoleptic characteristics. Only flower stigmas are used for its production, as the rest of the flower is discarded as waste. This fact represents a lack of sustainability, since around 230,000 flowers are necessary to produce 1 kg of saffron. The main aim of this study was to contribute to the valorization of Crocus sativus L. spice and its floral by-products, through the study of their nutritional value and composition, in terms of hydrophilic and lipophilic compounds, as well as their functional properties. The results showed that saffron stigmas and floral bio-residues presented high contents of fiber, and the most abundant macronutrient were the carbohydrates, followed by proteins, and a low content in fats. All samples had high concentrations of glucose, fructose, lactic and malic acids, and minerals, mainly K, Ca and Mg. Furthermore, the polyunsaturated fatty acids were predominant, being linoleic acid (C18:2n6) the most abundant. Therefore, this research provides more in-depth information about the composition of saffron stigmas and floral by-products, to be considered as promising sources for the development of functional ingredients with new applications in the food industry.

Influence of Synthetic Fibers on the Flexural Properties of Concrete: Prediction of Toughness as a Function of Volume, Slenderness Ratio and Elastic Modulus of Fibers.

The construction industry requires concrete with adequate post-cracking behavior for applications such as tunnels, bridges, and pavements. For this reason, polypropylene macrofibers are used, which are synthetic fibers that fulfill the function of providing residual strength to concrete. In this study, an experimental plan is carried out to evaluate the bending behavior of concrete reinforced with polypropylene fibers using the four-point bending test according to ASTM C1609. Three fiber dosages (3.6, 7.2 and 10.8 kg/m3) and three fiber lengths (40, 50, and 60 mm) were used. The use of macro polypropylene fibers increased the post-cracking behavior of concrete. In addition, based on the experimentally obtained results and available literature data, a multivariable equation was developed to predict the concrete toughness as a function of the volume, slenderness, and modulus of elasticity of the fibers. A Pearson's correlation coefficient, r of 0.90, showed a strong correlation between the developed equation and the experimental data. From this equation, it was possible to determine the participation of the following parameters in calculating toughness. The participation or weight of the fiber's modulus of elasticity on the concrete's tenacity is 26%, the volume of the fiber is 39%, the slenderness is 19%, and the reinforcement index is 16%.

Use of an electromagnetic-guided device to assist with post-pyloric placement of a nasoenteral feeding tube: A systematic review and meta-analysis.

Background and study aims While endoscopic-guided placement (EGP) of a post-pyloric nasoenteral feeding tube may improve caloric intake and reduce the risk of bronchoaspiration, an electromagnetic-guided placement (EMGP) method may obviate the need for endoscopic procedures. Therefore, the primary aim of this study was to perform a systematic review and meta-analysis of randomized trials comparing the efficacy and safety of EMGP versus EGP of a post-pyloric feeding tube. Methods Protocolized searches were performed from the inception through January 2021 following PRISMA guidelines. Only randomized controlled trials were included comparing EMGP versus EGP. Study outcomes included: technical success (defined as appropriate post-pyloric positioning), tube and patient associated adverse events (AEs), time to enteral nutrition, procedure-associated cost, and procedure time. Pooled risk difference (RD) and mean difference (MD) were calculated using a fixed-effects model and heterogeneity evaluated using Higgins test (I 2 ). Results Four randomized trials (n = 536) were included. A total of 287 patients were included in the EMGP group and 249 patients in the EGP group. There was no difference between EMGP versus EGP regarding technical success, tube-related AEs, patient-related AEs, procedure time, and time in the right position. Time to enteral nutrition favored EMGP (MD: -134.37 [-162.13, -106.61]; I 2  = 35 %); with significantly decreased associated cost (MD: -127.77 ($) [-135.8-119.73]; I 2  = 0 %). Conclusions Based on this study, EMGP and EGP were associated with similar levels of technical success and safety as well as time to complete the procedure. Despite this, EMGP was associated with reduced cost and time to initiation of nutrition.

mTORC2 Is the Major Second Layer Kinase Negatively Regulating FOXO3 Activity.

Forkhead box O (FOXO) proteins are transcription factors involved in cancer and aging and their pharmacological manipulation could be beneficial for the treatment of cancer and healthy aging. FOXO proteins are mainly regulated by post-translational modifications including phosphorylation, acetylation and ubiquitination. As these modifications are reversible, activation and inactivation of FOXO factors is attainable through pharmacological treatment. One major regulatory input of FOXO signaling is mediated by protein kinases. Here, we use specific inhibitors against different kinases including PI3K, mTOR, MEK and ALK, and other receptor tyrosine kinases (RTKs) to determine their effect on FOXO3 activity. While we show that inhibition of PI3K efficiently drives FOXO3 into the cell nucleus, the dual PI3K/mTOR inhibitors dactolisib and PI-103 induce nuclear FOXO translocation more potently than the PI3Kδ inhibitor idelalisib. Furthermore, specific inhibition of mTOR kinase activity affecting both mTORC1 and mTORC2 potently induced nuclear translocation of FOXO3, while rapamycin, which specifically inhibits the mTORC1, failed to affect FOXO3. Interestingly, inhibition of the MAPK pathway had no effect on the localization of FOXO3 and upstream RTK inhibition only weakly induced nuclear FOXO3. We also measured the effect of the test compounds on the phosphorylation status of AKT, FOXO3 and ERK, on FOXO-dependent transcriptional activity and on the subcellular localization of other FOXO isoforms. We conclude that mTORC2 is the most important second layer kinase negatively regulating FOXO activity.

Novel Insight into the Volatile Profile and Antioxidant Properties of Crocus sativus L. Flowers.

The current production system of saffron spice generates hundreds of tons of waste. Thus, the aim of this study was to value both saffron and its floral by-products as a source of natural bioactive extracts, studying the in vitro antioxidant capacity, the composition of the volatile fraction by GC-MS/MS, and the determination of crocetins esters by HPLC-PDA. Saffron stigmas and floral by-products showed a high content of polyphenols and different antioxidant properties. Floral bio-residues (tepals, stamens, and styles) presented a high concentration of anthocyanins, and stigmas had high levels of flavonoids, ß-carotene, and total crocins. In stigmas, 25 different volatile components were found, with safranal the most relevant. Floral by-products volatile composition consisted of 55 compounds with varying amounts depending on the drying treatment; all the samples presented acetic acid, 2(5H)-furanone, and phenylethyl alcohol. Therefore, saffron stigmas and flower by-products represent a sustainable source of bioactive ingredients for innovative healthy food formulations.

A clinically compatible drug-screening platform based on organotypic cultures identifies vulnerabilities to prevent and treat brain metastasis.

We report a medium-throughput drug-screening platform (METPlatform) based on organotypic cultures that allows to evaluate inhibitors against metastases growing in situ. By applying this approach to the unmet clinical need of brain metastasis, we identified several vulnerabilities. Among them, a blood-brain barrier permeable HSP90 inhibitor showed high potency against mouse and human brain metastases at clinically relevant stages of the disease, including a novel model of local relapse after neurosurgery. Furthermore, in situ proteomic analysis applied to metastases treated with the chaperone inhibitor uncovered a novel molecular program in brain metastasis, which includes biomarkers of poor prognosis and actionable mechanisms of resistance. Our work validates METPlatform as a potent resource for metastasis research integrating drug-screening and unbiased omic approaches that is compatible with human samples. Thus, this clinically relevant strategy is aimed to personalize the management of metastatic disease in the brain and elsewhere.

Saffron bioactives crocin, crocetin and safranal: effect on oxidative stress and mechanisms of action.

Saffron (Crocus sativus L.) is used as a spice for its organoleptic characteristics related to its coloring and flavoring properties, and it has been also used in traditional medicine to treat various diseases. The main chemical components responsible for these properties are crocin, crocetin and safranal. These compounds have been shown to have a wide spectrum of biological activities, including several properties as antigenotoxic, antioxidant, anticancer, anti-inflammatory, antiatherosclerotic, antidiabetic, hypotensive, hypoglycemic, antihyperlipidemic, antidegenerative and antidepressant, among others. This review article highlights the antioxidant effects of these bioactive compounds to reduce reactive oxygen species (ROS) and the mechanisms of action involved, since there are a multitude of diseases related to oxidative stress and the generation of free radicals (FRs). Recent studies have shown that the effects of crocin, crocetin and safranal against oxidative stress include the reduction in lipid peroxidation (malondialdehyde [MDA] levels) and nitric oxide (NO) levels, and the increase in the levels of glutathione, antioxidant enzymes (superoxide dismutase [SOD], catalase (CAT) and glutathione peroxidase [GPx]) and thiol content. Therefore, due to the great antioxidant effects of these saffron compounds, it makes saffron a potential source of bioactive extracts for the development of bioactive ingredients, which can be used to produce functional foods.

Probiotic red quinoa drinks for celiacs and lactose intolerant people: study of functional, physicochemical and probiotic properties during fermentation and gastrointestinal digestion.

The consumption of probiotic foods is rather limited for many sectors of the population (vegans, lactose intolerant and celiacs). Therefore, it is necessary to offer functional alternatives for these sectors. Different red quinoa drinks were fermented with L. plantarum (QLPBB) and B. longum (QBLBB) at different times. Results showed that microbial concentration reached high levels (6-8 Log CFU/mL) after 6 h and probiotic viability was very high (6 Log CFU/mL) after gastrointestinal digestion (GD). However, QBLBB reached the best probiotic concentration at 24 h. From 6 h of fermentation, probiotic resistance to some antibiotics, especially B. longum, could have a great potential to restore the intestinal microbiota during and after treatment with certain antibiotics. QLPBB showed the highest levels of total polyphenols and antioxidant capacity (AOC) after GD. Therefore, these red quinoa drinks have potential as functional probiotic beverages for vegans, celiacs and allergic to milk protein and lactose-intolerant people.

Screening Health-Promoting Compounds for Their Capacity to Induce the Activity of FOXO3.

Several chemical compounds including natural products have been suggested as being effective against age-related diseases or as beneficial for a healthy life. On the other hand, forkhead box O (FOXO) proteins are emerging as key cellular components associated with extreme human longevity. FOXO proteins are mainly regulated by posttranslational modifications and as these modifications are reversible, activation and inactivation of FOXO are attainable through pharmacological treatment. Here, we questioned whether a panel of compounds with known health-beneficial properties has the capacity to induce the activity of FOXO factors. We show that resveratrol, a phytoalexin present in grapes and other food products, the amide alkaloid piperlongumine found in the fruit of the long pepper, and the plant-derived ß-carboline compound harmine induced nuclear translocation of FOXO3. We also show that piperlongumine and harmine but not resveratrol activate FOXO-dependent transcription. We determined the half maximal effective concentration (EC50) values for resveratrol, piperlongumine, and harmine for FOXO translocation, and analyzed their inhibitory impact on chromosomal maintenance 1 (CRM1)-mediated nuclear export and the production of reactive oxygen species (ROS). We also used chemical biology approach and Western blot analysis to explore the underlying molecular mechanisms. We show that harmine, piperlongumine, and resveratrol activate FOXO3 independently of phosphoinositide 3-kinase (PI3K)/AKT signaling and the CRM1-mediated nuclear export. The effect of harmine on FOXO3 activity is at least partially mediated through the inhibition of dual-specificity tyrosine (Y) phosphorylationregulated kinase 1A (DYRK1A) and can be reverted by the inhibition of sirtuins (SIRTs).

Macrocyclization as a Source of Desired Polypharmacology. Discovery of Triple PI3K/mTOR/PIM Inhibitors.

The PI3K/AKT/mTOR and PIM kinase pathways contribute to the development of several hallmarks of cancer. Cotargeting of these pathways has exhibited promising synergistic therapeutic effects in liquid and solid tumor types. To identify molecules with combined activities, we cross-screened our collection of PI3K/(±mTOR) macrocycles (MCXs) and identified the MCX thieno[3,2-d]pyrimidine derivative 2 as a moderate dual PI3K/PIM-1 inhibitor. We report the medicinal chemistry exploration and biological characterization of a series of thieno[3,2-d]pyrimidine MCXs, which led to the discovery of IBL-302 (31), a potent, selective, and orally bioavailable triple PI3K/mTOR/PIM inhibitor. IBL-302, currently in late preclinical development (AUM302), has recently demonstrated efficacy in neuroblastoma and breast cancer xenografts. Additionally, during the course of our experiments, we observed that macrocyclization was essential to obtain the desired multitarget profile. As a matter of example, the open precursors 35-37 were inactive against PIM whereas MCX 28 displayed low nanomolar activity.

EUS-guided gastroenterostomy versus duodenal stent placement and surgical gastrojejunostomy for the palliation of malignant gastric outlet obstruction: a systematic review and meta-analysis.

PURPOSE: Malignant gastric outlet obstruction (GOO) is associated with significant morbidity and decreased quality of life, thereby necessitating effective and safe palliative treatment. As such, we sought to compare endoscopic ultrasound-guided gastroenterostomy (EUS-GE) versus duodenal stent (DS) placement and surgical gastrojejunostomy (SGJ) for palliation of malignant GOO. METHODS: Searches of electronic databases were performed to identify studies comparing EUS-GE versus DS and/or SGJ for palliative treatment of GOO. Outcomes included technical and clinical success, severe adverse events (SAEs), rate of stent obstruction (including tumor ingrowth), length of hospital stay (LOS), reintervention, and 30-day all-cause mortality. Differences in dichotomous and continuous outcomes were reported as risk difference and mean difference, respectively. RESULTS: Seven studies (n = 513 patients) were included. When compared to DS placement, EUS-GE was associated with a higher clinical success, fewer SAEs, decreased stent obstruction, lower rate of tumor ingrowth, and decreased need for reintervention. Compared to SGJ, EUS-GE was associated with a lower technical success; however, LOS was significantly decreased. All other outcomes including clinical success, SAEs, reintervention rate, and 30-day mortality were not significantly different between an EUS-guided versus surgical approach. CONCLUSIONS: EUS-GE was associated with significantly improved outcomes compared to DS placement for palliative treatment of malignant GOO. Despite SGJ possessing a higher technical success compared to EUS-GE, LOS was significantly longer with no difference in clinical success or rate of adverse events.

Pain relief in chronic pancreatitis: endoscopic or surgical treatment? a systematic review with meta-analysis.

BACKGROUND AND AIMS: Pain is one of the consequences of chronic pancreatitis (CP) that has the greatest impact on the quality of life of patients. Endoscopic and surgical interventions, by producing a decrease in intraductal pancreatic pressure, can provide pain relief. This is the first systematic review that includes only randomized clinical trials (RTCs) comparing outcomes in the short-term (less than 2 years) and long-term (more than 2 years) between these two types of interventions. MATERIAL AND METHODS: A comprehensive search of multiple electronic databases to identify RTCs comparing short and long-term pain relief, procedural complications, and days of hospitalization between endoscopic and surgical interventions was performed following the PRISMA guidelines. RESULTS: Three RCTs evaluating a total of 199 patients (99 in the endoscopy group and 100 in the surgery group) were included in this study. Surgical interventions provided complete pain relief, with statistical difference, in the long-term (16,4% vs 35.7%; RD 0.19; 95% CI 0.03-0.35; p = 0.02; I2 = 0%), without significant difference in short-term (17.5% vs 31.2%; RD 0.14; 95% CI -0.01-0.28; p = 0.07; I2 = 0%) when compared to endoscopy. There was no statistical difference in short-term (17.5% vs 28.1%; RD 0.11; 95% CI -0.04-0.25; p = 0.15; I2 = 0%) and long-term (34% vs 41.1%; RD 0.07; 95% CI -0.10-0.24; p = 0.42; I2 0%) in partial relief of pain between both interventions. In the short-term, both complications (34.9% vs 29.7%; RD 0.05; 95% CI -0.10-0.21; p = 0.50; I2 = 48%) and days of hospitalization (MD -1.02; 95% CI -2.61-0.58; p = 0.21; I2 = 0%) showed no significant differences. CONCLUSION: Surgical interventions showed superior results when compared to endoscopy in terms of complete long-term pain relief. The number of complications and length of hospitalization in both groups were similar.

Omipalisib inspired macrocycles as dual PI3K/mTOR inhibitors.

Activation of the phosphatidylinositol 3-kinase (PI3K)/mammalian target of rapamycin (mTOR) signaling pathway occurs frequently in a wide range of human cancers and is a main driver of cell growth, proliferation, survival, and chemoresistance of cancer cells. Compounds targeting this pathway are under active development as anticancer therapeutics and some of them have reached advanced clinical trials or been approved by the FDA. Dual PI3K/mTOR inhibitors combine multiple therapeutic efficacies in a single molecule by inhibiting the pathway both upstream and downstream of AKT. Herein, we report our efforts on the exploration of novel small molecule macrocycles (MCXs) as dual PI3K/mTOR inhibitors. Macrocyclization is an attractive approach used in drug discovery, as the semi-rigid character of these structures could provide improved potency, selectivity and favorable pharmacokinetic properties. Importantly, this strategy allows access to new chemical space thus obtaining a better intellectual property position. A series of MCXs based on GSK-2126458, a known clinical PI3K/mTOR inhibitor is described. These molecules showed potent biochemical and cellular dual PI3K/mTOR inhibition, demonstrated strong antitumoral effects in human cancer cell lines, and displayed good drug-like properties. Among them, MCX 83 presented remarkable selectivity against a panel of 468 kinases, high in vitro metabolic stability, and favorable pharmacokinetic parameters without significant CYP450 and h-ERG binding inhibition. This profile qualified this compound as a suitable candidate for future in vivo PK-PD and efficacy studies in mouse cancer models.

Global hyperactivation of enhancers stabilizes human and mouse naive pluripotency through inhibition of CDK8/19 Mediator kinases.

Pluripotent stem cells (PSCs) transition between cell states in vitro, reflecting developmental changes in the early embryo. PSCs can be stabilized in the naive state by blocking extracellular differentiation stimuli, particularly FGF-MEK signalling. Here, we report that multiple features of the naive state in human and mouse PSCs can be recapitulated without affecting FGF-MEK signalling or global DNA methylation. Mechanistically, chemical inhibition of CDK8 and CDK19 (hereafter CDK8/19) kinases removes their ability to repress the Mediator complex at enhancers. CDK8/19 inhibition therefore increases Mediator-driven recruitment of RNA polymerase II (RNA Pol II) to promoters and enhancers. This efficiently stabilizes the naive transcriptional program and confers resistance to enhancer perturbation by BRD4 inhibition. Moreover, naive pluripotency during embryonic development coincides with a reduction in CDK8/19. We conclude that global hyperactivation of enhancers drives naive pluripotency, and this can be achieved in vitro by inhibiting CDK8/19 kinase activity. These principles may apply to other contexts of cellular plasticity.

Induction of Lysosome Membrane Permeabilization as a Therapeutic Strategy to Target Pancreatic Cancer Stem Cells.

Despite significant efforts to improve pancreatic ductal adenocarcinoma (PDAC) clinical outcomes, overall survival remains dismal. The poor response to current therapies is partly due to the existence of pancreatic cancer stem cells (PaCSCs), which are efficient drivers of PDAC tumorigenesis, metastasis and relapse. To find new therapeutic agents that could efficiently kill PaCSCs, we screened a chemical library of 680 compounds for candidate small molecules with anti-CSC activity, and identified two compounds of a specific chemical series with potent activity in vitro and in vivo against patient-derived xenograft (PDX) cultures. The anti-CSC mechanism of action of this specific chemical series was found to rely on induction of lysosomal membrane permeabilization (LMP), which is likely associated with the increased lysosomal mass observed in PaCSCs. Using the well characterized LMP-inducer siramesine as a tool molecule, we show elimination of the PaCSC population in mice implanted with tumors from two PDX models. Collectively, our approach identified lysosomal disruption as a promising anti-CSC therapeutic strategy for PDAC.

Pyrido[2,3-b][1,5]benzoxazepin-5(6H)-one derivatives as CDK8 inhibitors.

CDK8 is a cyclin-dependent kinase that forms part of the mediator complex, and modulates the transcriptional output from distinct transcription factors involved in oncogenic control. Overexpression of CDK8 has been observed in various cancers, representing a potential target for developing novel CDK8 inhibitors in cancer therapeutics. In the course of our investigations to discover new CDK8 inhibitors, we designed and synthesized tricyclic pyrido[2,3-b][1,5]benzoxazepin-5(6H)-one derivatives, by introduction of chemical complexity in the multi-kinase inhibitor Sorafenib taking into account the flexibility of the P-loop motif of CDK8 protein observed after analysis of structural information of co-crystallized CDK8 inhibitors. In vitro evaluation of the inhibitory activity of the prepared compounds against CDK8 led us to identify compound 2 as the most potent inhibitor of the series (IC50 = 8.25 nM). Co-crystal studies and the remarkable selectivity profile of compound 2 are presented. Compound 2 showed moderate reduction of phosphorylation of CDK8 substrate STAT1 in cells, in line with other reported Type II CDK8 inhibitors. We propose herein an alternative to find a potential therapeutic use for this chemical series.

Non-Dairy Fermented Beverages as Potential Carriers to Ensure Probiotics, Prebiotics, and Bioactive Compounds Arrival to the Gut and Their Health Benefits.

In alignment with Hippocrates' aphorisms "Let food be your medicine and medicine be your food" and "All diseases begin in the gut", recent studies have suggested that healthy diets should include fermented foods to temporally enhance live microorganisms in our gut. As a result, consumers are now demanding this type of food and fermented food has gained popularity. However, certain sectors of population, such as those allergic to milk proteins, lactose intolerant and strict vegetarians, cannot consume dairy products. Therefore, a need has arisen in order to offer consumers an alternative to fermented dairy products by exploring new non-dairy matrices as probiotics carriers. Accordingly, this review aims to explore the benefits of different fermented non-dairy beverages (legume, cereal, pseudocereal, fruit and vegetable), as potential carriers of bioactive compounds (generated during the fermentation process), prebiotics and different probiotic bacteria, providing protection to ensure that their viability is in the range of 106-107 CFU/mL at the consumption time, in order that they reach the intestine in high amounts and improve human health through modulation of the gut microbiome.