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In this article, the author discusses his book "Frankenstein, or the 8­bit Prometheus" available in 2017 through Chili Com Carne and Thisco , and featuring music­related essays incorporating hyper mash­up, gabber and break­core.

The aim was to study the shelf-life over 18 months of storage (no light and oxygen exposure) of chili peppers flavoured olive oils comparing flavouring methods of co-milling of fresh chili peppers with sound olives at mill scale with temporary and permanent infusion of dried chili peppers in olive oil. Tocopherols, secoiridoids and capsaicinoids by HPLC-DAD, volatile compounds by HS-SPME-GC-MS, and sensory profiles were studied. The decrease in tocopherols and secoiridoids was greater in "infusion" samples, while a significant increase in capsaicinoids was observed in "permanent infusion" samples. The main changes were observed for sensory and volatile profiles: "infusion" samples were defective already after 2 months with significant increase of defects-relating volatile compounds, while "co-milling" samples were defects-free and characterized by nice balance among hotness/heat and pepper fruity/taste during entire storage. "Co-milling" samples showed better shelf-life (18 months) than infusion ones, and even than EVOO control samples (12 months)., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

A head space-solid phase microextraction-gas chromatography-mass spectrometery (HS-SPME-GC-MS) method for the simultaneous analysis of pentene dimers from lipoxygenase (LOX) pathway, monoterpenes, and sesquiterpenes in extra virgin olive oil (EVOO) was proposed. A Doehlert design was performed; the conditions of the HS-SPME preconcentration step (extraction temperature, extraction time, sample amount, and desorption time) were optimized by response surface methodology, allowing defining the method operable design region. A quantitative method was set up using the multiple internal standard normalization approach: four internal standards were used, and the most suitable one was selected for area normalization of each external standard. The quantitative method was successfully validated and applied to a series of monocultivar EVOOs. This is the first paper in which a quantitative method using commercial standards has been proposed for the analysis of an important class of molecules of EVOO such as pentene dimers. The optimized method is suitable for routine analysis aimed at characterizing high quality EVOOs.

Neoadjuvant immunotherapy plus chemotherapy improves event-free survival (EFS) and pathologic complete response (0% residual viable tumor (RVT) in primary tumor (PT) and lymph nodes (LNs)), and is approved for treatment of resectable lung cancer. Pathologic response assessment after neoadjuvant therapy is the potential analog to radiographic response for advanced disease. However, %RVT thresholds beyond pathologic complete response and major pathologic response (≤10% RVT) have not been explored. Pathologic response was prospectively assessed in the randomized, phase 3 CheckMate 816 trial (NCT02998528), which evaluated neoadjuvant nivolumab (anti-programmed death protein 1) plus chemotherapy in patients with resectable lung cancer. RVT, regression and necrosis were quantified (0-100%) in PT and LNs using a pan-tumor scoring system and tested for association with EFS in a prespecified exploratory analysis. Regardless of LN involvement, EFS improved with 0% versus >0% RVT-PT (hazard ratio = 0.18). RVT-PT predicted EFS for nivolumab plus chemotherapy (area under the curve = 0.74); 2-year EFS rates were 90%, 60%, 57% and 39% for patients with 0-5%, >5-30%, >30-80% and >80% RVT, respectively. Each 1% RVT associated with a 0.017 hazard ratio increase for EFS. Combining pathologic response from PT and LNs helped differentiate outcomes. When compared with radiographic response and circulating tumor DNA clearance, %RVT best approximated EFS. These findings support pathologic response as an emerging survival surrogate. Further assessment of the full spectrum of %RVT in lung cancer and other tumor types is warranted. ClinicalTrials.gov registration: NCT02998528 ., (© 2023. The Author(s).)

Introduction: A small percentage of patients with SCLC experience durable responses to immune checkpoint blockade (ICB). Defining determinants of immune response may nominate strategies to broaden the efficacy of immunotherapy in patients with SCLC. Prior studies have been limited by small numbers or concomitant chemotherapy administration., Methods: CheckMate 032, a multicenter, open-label, phase 1/2 trial evaluating nivolumab alone or with ipilimumab was the largest study of ICB alone in patients with SCLC. We performed comprehensive RNA sequencing of 286 pretreatment SCLC tumor samples, assessing outcome on the basis of defined SCLC subtypes (SCLC-A, -N, -P, and -Y), and expression signatures associated with durable benefit, defined as progression-free survival more than or equal to 6 months. Potential biomarkers were further explored by immunohistochemistry., Results: None of the subtypes were associated with survival. Antigen presentation machinery signature (p = 0.000032) and presence of more than or equal to 1% infiltrating CD8+ T cells by immunohistochemistry (hazard ratio = 0.51, 95% confidence interval: 0.27-0.95) both correlated with survival in patients treated with nivolumab. Pathway enrichment analysis revealed the association between durable benefit from immunotherapy and antigen processing and presentation. Analysis of epigenetic determinants of antigen presentation identified LSD1 gene expression as a correlate of worse survival outcomes for patients treated with either nivolumab or the combination of nivolumab and ipilimumab., Conclusions: Tumor antigen processing and presentation is a key correlate of ICB efficacy in patients with SCLC. As antigen presentation machinery is frequently epigenetically suppressed in SCLC, this study defines a targetable mechanism by which we might improve clinical benefit of ICB for patients with SCLC., (Copyright © 2023 International Association for the Study of Lung Cancer. Published by Elsevier Inc. All rights reserved.)

Proso millet has been proposed as an effective anti-diabetic food thanks to the combined action of polyphenols and starch. This study aimed to characterize the chemical composition of twenty-five accessions, in order to enhance this cereal as an alternative to available starch for food applications or to propose new food ingredients with health benefits. Proso millet contained a high percentage of starch, reaching values of 58.51%. The amylose content showed high variability, with values ranging from 1.36 to 42.70%, and significantly higher values were recorded for the white accessions than for the colored ones. High-resistant starch content (13.41-26.07%) was also found. The HPLC-MS analysis showed the same phenolic pattern in all the samples. Cinnamic acids are the most abundant compounds and significant differences in their total content were found (0.69 to 1.35 mg/g DW), while flavonoids were only detected in trace amounts. Statistical results showed significantly higher antiradical activity in the colored accessions than in the white ones.

The tumor immune composition influences prognosis and treatment sensitivity in lung cancer. The presence of effective adaptive immune responses is associated with increased clinical benefit after immune checkpoint blockers. Conversely, immunotherapy resistance can occur as a consequence of local T-cell exhaustion/dysfunction and upregulation of immunosuppressive signals and regulatory cells. Consequently, merely measuring the amount of tumor-infiltrating lymphocytes (TILs) may not accurately reflect the complexity of tumor-immune interactions and T-cell functional states and may not be valuable as a treatment-specific biomarker. In this work, we investigate an immune-related biomarker (PhenoTIL) and its value in associating with treatment-specific outcomes in non-small cell lung cancer (NSCLC). PhenoTIL is a novel computational pathology approach that uses machine learning to capture spatial interplay and infer functional features of immune cell niches associated with tumor rejection and patient outcomes. PhenoTIL's advantage is the computational characterization of the tumor immune microenvironment extracted from H&E-stained preparations. Association with clinical outcome and major non-small cell lung cancer (NSCLC) histology variants was studied in baseline tumor specimens from 1,774 lung cancer patients treated with immunotherapy and/or chemotherapy, including the clinical trial Checkmate 057 (NCT01673867)., (© 2023. The Author(s).)

In the context of the olive oil flavoured with chili peppers, the aim of this study was to compare co-milling of sound olives and fresh chili peppers at mill scale to infusion of dried chili peppers in oil, using the same batch of olives for all oils. Capsaicinoids by HPLC-DAD, volatile profile by HS-SPME-GC-MS and HS-GC-IMS and sensory profile were characterized. Capsaicinoids were statistically higher in oils prepared with green (52.0-68.0 mg/kg) than red (48.0-60.2 mg/kg) chili peppers. Oils flavoured by infusion showed higher contents of volatile compounds linked to defects such as acetic acid, with winey/vinegary sensory defect (median, 1.72-2.02) and no fresh pepper flavour. Oils prepared by co-milling resulted rich in the typical esters of chili pepper (6.175 and 4.156 mg/kg with green and red chili peppers, respectively), with pleasant hotness sensation and fresh pepper flavour. Overall, the co-milling approach allowed obtaining flavoured samples with improved sensory quality., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022. Published by Elsevier Ltd.)

The effect of industrial drying processes on phenols and polysaccharides of olive pomace (pâté) and pomegranate peel was studied, with the aim to re-use pomegranate and olive oil by-products. Pomegranate peel (Wonderful and G1 varieties) was oven-dried at different temperatures, taking into account peel thickness and size. Pâté was freeze-dried and oven dried at 50-110 °C, at lab scale; then, an industrial drying system (150 °C) was compared to freeze-drying. All dried samples were analyzed in terms of phenolic and polysaccharides compounds. Drying at room temperature of small pieces of pomegranate peel guaranteed the highest humidity removal and recovery of phenols. Sugar analysis, DLS and 1 H NMR confirmed that polysaccharide fractions were not significantly affected by the highest drying temperatures (42 °C for pomegranate, 150 °C for pâté). The two drying procedures at large scale were suitable for avoiding degradation of phenols, maintaining the same profiles of the corresponding freeze-dried samples., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Millet is the sixth-highest yielding grain in the world and a staple crop for millions of people. Fermentation was applied in this study to improve the nutritional properties of pearl millet. Three microorganism combinations were tested: Saccharomyces boulardii (FPM1), Saccharomyces cerevisiae plus Campanilactobacillus paralimentarius (FPM2) and Hanseniaspora uvarum plus Fructilactobacillus sanfranciscensis (FPM3). All the fermentation processes led to an increase in minerals. An increase was observed for calcium: 254 ppm in FPM1, 282 ppm in FPM2 and 156 ppm in the unfermented sample. Iron increased in FPM2 and FPM3 (approx. 100 ppm) with respect the unfermented sample (71 ppm). FPM2 and FPM3 resulted in richer total phenols (up to 2.74 mg/g) compared to the unfermented sample (2.24 mg/g). Depending on the microorganisms, it was possible to obtain different oligopeptides with a mass cut off ≤10 kDalton that was not detected in the unfermented sample. FPM2 showed the highest resistant starch content (9.83 g/100 g) and a prebiotic activity on Bifidobacterium breve B632, showing a significant growth at 48 h and 72 h compared to glucose ( p < 0.05). Millet fermented with Saccharomyces cerevisiae plus Campanilactobacillus paralimentarius can be proposed as a new food with improved nutritional properties to increase the quality of the diet of people who already use millet as a staple food.

Polysaccharides from pomegranate peel (Wonderful and Purple Queen® varieties) were extracted by hot water and fractionated using ethanol. Three fractions (F1-F2-F3) were obtained for each sample. Polysaccharides' yield was higher for Purple Queen®: 13% dw. Polysaccharides of the three fractions were characterized by size exclusion chromatography (SEC), dynamic light scattering (DLS), 1 H NMR, methylation, and acylation degree. Differently from SEC, DLS highlighted some differences between the polysaccharides's molecular sizes of the fractions. The highest methylation and acylation degree was observed for F3 of Purple Queen®: 74.0% and 18.6%, respectively. The percentage of galacturonic acid confirmed the presence of pectin in almost all the fractions recognized as homogalacturonan. Arabinan and arabinogalactan were also found in all the collected F3 samples, although in different proportions. The stepwise fractionation process followed by a multi-methodological analytical investigation was helpful to improve the knowledge of the pectic polysaccharides of pomegranate., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The study analysed polysaccharides and phenolic compounds in widely consumed but little studied date fruits varieties such as Sukkari, Ajwa, Segae, Barrny and Khalas harvested at Tamr stage. The total phenols were in similar amount in the five varieties and ranged from 20 to 50 mg/100 g DW. The decoction and successive centrifugation made it possible to collect two main polysaccharide fractions for all the selected fruits. For each variety the first fraction was more abundant, with a lower swelling capacity and a higher amount of galacturonic acid (28.3% to 40.1%). The second fraction was only soluble in alkaline solution, with an average galacturonic acid content of only 17%. The different structure of the two polysaccharide fractions was also confirmed by the composition in neutral sugars and the degrees of methylation and acetylation. The proposed extraction procedure could be applied for larger scale extraction of the date fruit polysaccharides., (Copyright © 2022. Published by Elsevier Ltd.)

Climate change has led to rediscovery of minor drought tolerant grains such as millet. Among its bioactive molecules, steryl ferulates have been poorly explored. Steryl ferulates composition of was investigated by high performance liquid chromatography-diode array detector-tandem mass spectrometry and high resolution tandem mass spectrometry in twenty-two millet samples and also in some fermented and microwave heated samples. Six compounds were found in Panicum, Pennisetum, Eleusine and Setaria genera, with a prevalence of campestanyl and sitostanyl ferulate. The lowest steryl ferulates content was found in Panicum, with values ranging from 2.98 ± 0.04 µg/g to 8.72 ± 0.41 µg/g. Foxtail millet and finger millet showed the highest amount with 46.07 ± 5.20 µg/g and 85.29 ± 4.30 µg/g, respectively. As for pearl millet, microwave heating and fermentation increased steryl ferulates by two (33.77 ± 0.88 µg/g) and five (75.83 ± 1.25 µg/g) times, with respect to the untreated sample. Microwave heating and fermentation could be used to increase steryl ferulates in millet., (Copyright © 2022. Published by Elsevier Ltd.)

Purpose: Immune checkpoint blockade (ICB) in conjunction with chemotherapy is approved for the treatment of extensive-stage small-cell lung cancer (SCLC). Although specific genomic abnormalities such as KEAP1 and STK11 gene mutations are associated with resistance to ICB in non-SCLC, no genomic abnormality has been found in association with resistance to ICB in SCLC., Materials and Methods: We first analyzed a retrospective cohort of 42 patients with SCLC treated with single-agent ICB or ICB combination (data set A). We then validated our results in a large prospective clinical trial of 460 patients (CheckMate 032, data set B). DNA and RNA sequencing were performed., Results: In data set A, patients treated with ICB with RB1 wild-type (WT) had a median overall survival (OS) of 23.1 months (95% CI, 9 to 37.5), whereas the RB1 mutant OS was 5 months (95% CI, 2.5 to 26; P = .04). Differentially expressed gene analysis between RB1 mutant and RB1 WT samples indicated the enrichment of downregulated immune-related genes and an immune exclusion phenotype among RB1 mutant but not in the RB1 WT tumor samples. We then assessed results from 460 patients enrolled in CheckMate 032, a trial of nivolumab (NIVO) or NIVO + ipilimumab only in SCLC. In this large cohort, RB1 WT patients had significantly improved outcome with NIVO therapy compared with mutant patients (hazard ratio, 1.41; 95% CI, 1.02 to 2.01; P = .041). High RB1 loss-of-function (LOF) signature scores significantly associated with neuroendocrine subtypes (ASCL1 and NeuroD1). However, neuroendocrine subtypes did not associate with OS. Remarkably, patients with lower RB1 LOF scores had longer OS following treatment with NIVO., Conclusion: SCLC patients with RB1 WT status or lower RB1 LOF signature scores by transcriptomics have better outcomes with ICB monotherapy., Competing Interests: Afshin DowlatiConsulting or Advisory Role: AbbVie/Stemcentrx, AstraZeneca, Bristol Myers Squibb, Takeda, Bayer, G1 Therapeutics, Ipsen, MerckResearch Funding: Lilly/ImClone (Inst), Amgen (Inst), Bristol Myers Squibb (Inst), EMD Serono (Inst), Tesaro (Inst), Takeda (Inst), Mirati Therapeutics (Inst), AbbVie/Stemcentrx (Inst), United Therapeutics (Inst), Regeneron (Inst), Bayer (Inst), Seattle Genetics (Inst), Symphogen (Inst), Ipsen (Inst) Timothy ChanLeadership: Cancer Genetics, Illumina, Bristol Myers SquibbStock and Other Ownership Interests: Gritstone BioHonoraria: IlluminaConsulting or Advisory Role: Bristol Myers Squibb/Celgene, Illumina, LG Chem, PfizerResearch Funding: Bristol Myers Squibb (Inst), AstraZeneca/MedImmune (Inst), Pfizer (Inst), Nysnobio (Inst)Patents, Royalties, Other Intellectual Property: Neoantigen discovery and genomic biomarkers for immunotherapy response Brian HenickThis author is a member of the JCO Precision Oncology Editorial Board. Journal policy recused the author from having any role in the peer review of this manuscript.Consulting or Advisory Role: AstraZeneca, IDEAYA Biosciences, Jazz PharmaceuticalsResearch Funding: NexImmune Xuya WangEmployment: Bristol Myers SquibbStock and Other Ownership Interests: Bristol Myers Squibb Parul DoshiEmployment: Bristol Myers Squibb, Gilead SciencesStock and Other Ownership Interests: Bristol Myers SquibbTravel, Accommodations, Expenses: Bristol Myers Squibb, Gilead Sciences Jyoti PatelConsulting or Advisory Role: AbbVie, AstraZeneca, Takeda Science Foundation, Lilly, GenentechResearch Funding: Bristol Myers Squibb (Inst) Fengshen KuoStock and Other Ownership Interests: Sanofi, General Electric, 10x Genomics, Merck, Cigna, Cigna, Viatris, Pfizer Han ChangEmployment: Bristol Myers SquibbStock and Other Ownership Interests: Bristol Myers SquibbResearch Funding: Bristol Myers SquibbPatents, Royalties, Other Intellectual Property: I am an employee of BMS, and an inventor on one or more pending pa tent applications, including applications for TMB as a predictive biomarker of immunotherapyTravel, Accommodations, Expenses: Bristol Myers Squibb David BalliEmployment: Bristol Myers SquibbStock and Other Ownership Interests: Bristol Myers SquibbNo other potential conflicts of interest were reported.

Despite known histological, biological, and clinical differences between lung adenocarcinoma (LUAD) and squamous cell carcinoma (LUSC), relatively little is known about the spatial differences in their corresponding immune contextures. Our study of over 1000 LUAD and LUSC tumors revealed that computationally derived patterns of tumor-infiltrating lymphocytes (TILs) on H&E images were different between LUAD (N = 421) and LUSC (N = 438), with TIL density being prognostic of overall survival in LUAD and spatial arrangement being more prognostically relevant in LUSC. In addition, the LUAD-specific TIL signature was associated with OS in an external validation set of 100 NSCLC treated with more than six different neoadjuvant chemotherapy regimens, and predictive of response to therapy in the clinical trial CA209-057 (n = 303). In LUAD, the prognostic TIL signature was primarily comprised of CD4 + T and CD8 + T cells, whereas in LUSC, the immune patterns were comprised of CD4 + T, CD8 + T, and CD20 + B cells. In both subtypes, prognostic TIL features were associated with transcriptomics-derived immune scores and biological pathways implicated in immune recognition, response, and evasion. Our results suggest the need for histologic subtype-specific TIL-based models for stratifying survival risk and predicting response to therapy. Our findings suggest that predictive models for response to therapy will need to account for the unique morphologic and molecular immune patterns as a function of histologic subtype of NSCLC., (© 2022. The Author(s).)

Background: Neoadjuvant or adjuvant chemotherapy confers a modest benefit over surgery alone for resectable non-small-cell lung cancer (NSCLC). In early-phase trials, nivolumab-based neoadjuvant regimens have shown promising clinical activity; however, data from phase 3 trials are needed to confirm these findings., Methods: In this open-label, phase 3 trial, we randomly assigned patients with stage IB to IIIA resectable NSCLC to receive nivolumab plus platinum-based chemotherapy or platinum-based chemotherapy alone, followed by resection. The primary end points were event-free survival and pathological complete response (0% viable tumor in resected lung and lymph nodes), both evaluated by blinded independent review. Overall survival was a key secondary end point. Safety was assessed in all treated patients., Results: The median event-free survival was 31.6 months (95% confidence interval [CI], 30.2 to not reached) with nivolumab plus chemotherapy and 20.8 months (95% CI, 14.0 to 26.7) with chemotherapy alone (hazard ratio for disease progression, disease recurrence, or death, 0.63; 97.38% CI, 0.43 to 0.91; P = 0.005). The percentage of patients with a pathological complete response was 24.0% (95% CI, 18.0 to 31.0) and 2.2% (95% CI, 0.6 to 5.6), respectively (odds ratio, 13.94; 99% CI, 3.49 to 55.75; P<0.001). Results for event-free survival and pathological complete response across most subgroups favored nivolumab plus chemotherapy over chemotherapy alone. At the first prespecified interim analysis, the hazard ratio for death was 0.57 (99.67% CI, 0.30 to 1.07) and did not meet the criterion for significance. Of the patients who underwent randomization, 83.2% of those in the nivolumab-plus-chemotherapy group and 75.4% of those in the chemotherapy-alone group underwent surgery. Grade 3 or 4 treatment-related adverse events occurred in 33.5% of the patients in the nivolumab-plus-chemotherapy group and in 36.9% of those in the chemotherapy-alone group., Conclusions: In patients with resectable NSCLC, neoadjuvant nivolumab plus chemotherapy resulted in significantly longer event-free survival and a higher percentage of patients with a pathological complete response than chemotherapy alone. The addition of nivolumab to neoadjuvant chemotherapy did not increase the incidence of adverse events or impede the feasibility of surgery. (Funded by Bristol Myers Squibb; CheckMate 816 ClinicalTrials.gov number, NCT02998528.)., (Copyright © 2022 Massachusetts Medical Society.)

Anogeissus leiocarpus DC is an evergreen tree, widely distributed in Asia and Africa. The stem bark is used in traditional medicine, and as chewing sticks and infusion. Nowadays, it is becoming increasingly important to define the phytochemical profile of less studied edible plants. Aim of this research was a first complete characterization of ellagitannins and triterpenoids profiles by HPLC-DAD-MS and 1 H NMR and analyses. A total of 59 compounds were identified including 43 ellagitannins and 16 triterpenoids, mainly oleane derivatives and glycosylated forms. Among ellagitannins, roburin, vescalin and castalin were found for the first time. Tannins accounted for 38.9% whereas triterpenoids were 4.8%, both estimated on dry decoction. The decoction was preliminary tested against osteoarthritis in rats. The characterization of the main phytochemicals of Anogeissus leiocarpus DC stem bark decoction is a necessary step to evaluate nutraceutical properties, paving the way for possible food applications of this plant., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

The degree of metastatic disease varies widely among patients with cancer and affects clinical outcomes. However, the biological and functional differences that drive the extent of metastasis are poorly understood. We analyzed primary tumors and paired metastases using a multifluorescent lineage-labeled mouse model of pancreatic ductal adenocarcinoma (PDAC)-a tumor type in which most patients present with metastases. Genomic and transcriptomic analysis revealed an association between metastatic burden and gene amplification or transcriptional upregulation of MYC and its downstream targets. Functional experiments showed that MYC promotes metastasis by recruiting tumor-associated macrophages, leading to greater bloodstream intravasation. Consistent with these findings, metastatic progression in human PDAC was associated with activation of MYC signaling pathways and enrichment for MYC amplifications specifically in metastatic patients. Collectively, these results implicate MYC activity as a major determinant of metastatic burden in advanced PDAC. SIGNIFICANCE: Here, we investigate metastatic variation seen clinically in patients with PDAC and murine PDAC tumors and identify MYC as a major driver of this heterogeneity. This article is highlighted in the In This Issue feature, p. 275 ., (©2021 The Authors; Published by the American Association for Cancer Research.)

Phenolic and triterpenoid compounds of the olive tree are recognized as having a key role in health promotion, thanks to their multiple protective actions in humans. To expand the source of these bioactive compounds, the phenolic and triterpenoid profiles of leaf, branch, destoned fruit, destoned pomace, shell, seed, and extra virgin olive oil from the Frantoio , Leccino , and Moraiolo olive cultivars were simultaneously characterized by HPLC-DAD-MS. Overall, 43 molecules were quantitated and expressed on the obtained dry extracts. Oleuropein was mainly concentrated in branches (82.72 g/kg), fruits (55.79 g/kg), leaves (36.71 g/kg), and shells (1.26 g/kg), verbascoside (4.88 g/kg) in pomace, and nüzhenide 11-methyl oleoside (90.91 g/kg) in seeds. Among triterpenoids, which were absent in shells, the highest amount of oleanolic acid was found in olive leaves (11.88 g/kg). HCT-116 colorectal cells were chosen to assess the cytotoxicity of the dry extract, using the phytocomplex from Frantoio, which was the richest in phenols and triterpenoids. The IC 50 was also determined for 13 pure molecules (phenols and terpenoids) detected in the extracts. The greatest inhibition on the cell's proliferation was induced by the branch dry extract (IC 50 88.25 μg/mL) and by ursolic acid (IC 50 24 μM). A dose-dependent relationship was observed for the tested extracts.

Epithelial plasticity, or epithelial-to-mesenchymal transition (EMT), is a well-recognized form of cellular plasticity, which endows tumor cells with invasive properties and alters their sensitivity to various agents, thus representing a major challenge to cancer therapy. It is increasingly accepted that carcinoma cells exist along a continuum of hybrid epithelial-mesenchymal (E-M) states and that cells exhibiting such partial EMT (P-EMT) states have greater metastatic competence than those characterized by either extreme (E or M). We described recently a P-EMT program operating in vivo by which carcinoma cells lose their epithelial state through post-translational programs. Here, we investigate the underlying mechanisms and report that prolonged calcium signaling induces a P-EMT characterized by the internalization of membrane-associated E-cadherin (ECAD) and other epithelial proteins as well as an increase in cellular migration and invasion. Signaling through Gαq-associated G-protein-coupled receptors (GPCRs) recapitulates these effects, which operate through the downstream activation of calmodulin-Camk2b signaling. These results implicate calcium signaling as a trigger for the acquisition of hybrid/partial epithelial-mesenchymal states in carcinoma cells., (© 2021 The Authors.)

Wine and olive oil making by-products are rich sources of bioactive compounds suitable for new healthy recipes of staple foods. In this study, the profile of pasta (tagliatelle) fortified with 7% of grape pomace (GP) or olive pomace (pâté, OP) was studied, focusing on phenolic compounds after cooking. The enriched tagliatelle retained the same monoglycosylated and acetylated anthocyanins found in grape pomace. The fortified tagliatelle with a new milling by-product called pâté retained hydroxytyrosol after cooking (6.6 mg/100 g). In both the two types of enriched tagliatelle the fiber content increased of approx. 3%, while the added phenols retained after cooking by tagliatelle fortified with GP and OP were 6.21 mg/100 g and 9 mg/100 g, respectively. The fortified tagliatelle retained a good cooking resistance and a good texture after cooking, thus enhancing the nutritional profile of pasta, a staple food usually characterized by a negligible amount of phenolic compounds and fiber., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

The characterization of six varieties of native Andean potatoes with a wide biodiversity in tuber shape, flesh, and skin color was performed, through the determination of their proximate composition, mineral content, and phenolic profile. Minerals concentration revealed significant genotypic variation. Potassium was the most abundant element in all varieties, ranging from 7272.9 to 13,059.9 µg/g and from 12,418 to 17,388.6 µg/g dried weight for the flesh and skin samples, respectively. Iron content was relevant, ranging from 20.5 to 39.9 µg/g and from 112.2 to 288.8 µg/g dried weight in flesh and skin samples, respectively. Phenolic compounds were consistently higher in the skin than in the flesh. The total content varied greatly from 19.5 to 2015.3 µg/g and from 1592.3 to 14807.3 µg/g dried weight for flesh and skin tissues, respectively. 5-caffeoylquinic acid was 74% of the total phenolic acids. Different pattern of anthocyanins was found, depending on the color of the variety; the red genotypes contained predominantly pelargonidin derivatives, while the purple samples had petunidin as a major anthocyanidin. This study increases the knowledge of the composition of the local Andean varieties (which are only scarcely studied so far), helping to enhance these genotypes and the conservation of biodiversity.

Idiopathic pulmonary fibrosis (IPF) is a chronic disease with high mortality and is refractory to treatment. Pulmonary macrophages can both promote and repress fibrosis, however molecular mechanisms regulating macrophage functions during fibrosis remain poorly understood. FOXM1 is a transcription factor and is not expressed in quiescent lungs. Herein, we show that FOXM1 is highly expressed in pulmonary macrophages within fibrotic lungs of IPF patients and mouse fibrotic lungs. Macrophage-specific deletion of Foxm1 in mice (myFoxm1-/-) exacerbated pulmonary fibrosis. Inactivation of FOXM1 in vivo and in vitro increased p38 MAPK signaling in macrophages and decreased DUSP1, a negative regulator of p38 MAPK pathway. FOXM1 directly activated Dusp1 promoter. Overexpression of DUSP1 in FOXM1-deficient macrophages prevented activation of p38 MAPK pathway. Adoptive transfer of wild-type monocytes to myFoxm1-/- mice alleviated bleomycin-induced fibrosis. Altogether, contrary to known pro-fibrotic activities in lung epithelium and fibroblasts, FOXM1 has anti-fibrotic function in macrophages by regulating p38 MAPK., Competing Interests: The authors have declared that no competing interests exist.

Pomegranate is receiving renewed commercial and scientific interest, therefore a deeper knowledge of the chemical composition of the fruits of less studied varieties is required. In this work, juices from arils and decoctions from mesocarp plus exocarp were prepared from fifteen varieties. Samples were submitted to High Performance Liquid Chromatography-Diode Array Detector-Mass Spectrometry, spectrophotometric and colorimetric CIEL*a*b* analyses. Antioxidant, antiradical and metal chelating properties, inhibitory activity against tyrosinase and α-amylase enzymes were also evaluated. All varieties presented the same main phenols; anthocyanins and ellagitannins were widely variable among varieties, with the richest anthocyanin content in the juices from the Wonderful and Soft Seed Maule varieties (approx. 660 mg/L) and the highest ellagitannin content in the peel of the Black variety (approx. 133 mg/g dry matter). A good correlation was shown between the colour hue and the delphinidin/cyanidin ratio in juices ( R 2 = 0.885). Total polysaccharide yield ranged from 3% to 12% of the peels' dry weight, with the highest content in the Black variety. Decoctions (24.44-118.50 mg KAE/g) showed better in vitro antioxidant properties and higher inhibitory capacity against tyrosinase than juices (not active-16.56 mg KAE/g); the inhibitory capacity against α-amylase was similar and quite potent for juices and decoctions. Knowledge about the chemical composition of different pomegranate varieties will allow for a more aware use of the different parts of the fruit., Competing Interests: The authors declare no conflict of interest.

Millet is underutilized in Europe, despite its advantages compared to other common cereals. In Asia and Africa, millet is mainly eaten in fermented form; its consumption has beneficial properties on human health. Three millet batches were compared in terms of free and bound phenols by High Performance Liquid Chromatography-Diode Array Detector-Mass Spectrometry (HPLC-DAD-MS). The richest one in terms of bound phenols was selected for testing via a basic (0.1 M NaOH) and an acidic (1.2 M H 2 SO 4 ) hydrolysis, in which 149.3 and 193.6 mg/100 g of phenols were recovered, respectively. The ability of fermentation, with yeast and Lactobacilli , to increase the content of phenolic compounds was verified. Five withdrawalswere performed to verify the influence of fermentation time on the total phenolic content. The greatest phenolic content was observed after 72 h. Fermentation increased the cinnamic acids and flavonoids contents by approximately 30%. Vitexin and vitexin 2″- O -rhamnoside contents were significantly higher in the fermented millet; these compounds partially inhibit the protein tyrosine phosphatase enzyme, which is overexpressed in type-2 diabetes. A molecular dynamic simulation showed the two flavonoids to be allosteric inhibitors. The phenolic extract from fermented millet demonstrated a higher level of antioxidant protection on human erythrocytes by ex vivo cellular antioxidant activity in red blood cells. In this context, functional foods based on fermented millet could represent a new trend in European markets.

Extraction of free and bound phenols from millet in acidic and basic hydrolytic conditions were compared for the first time. Acidic hydrolysis was able to extract the highest amount of total phenolic compounds (up to 178 mg/100 g) while the basic hydrolysis underestimates the phenolic concentration. Our findings pointed out for the first time that methyl ferulate is naturally present as bound phenol in millet. Response Surface Methodology was then applied to both acidic and basic hydrolytic extractive conditions: the acidic procedure, optimized in terms of extractive time and temperature and concentration of the acidic mean, gave the best results, allowing definition of Method Operable Design Region and quantitation of the total amount of phenols in millet samples in a single extractive step. This optimized method is suitable for further accurate investigations of the typical phenols of the numerous varieties of this recently re-discovered minor cereal., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Fermented cereals, staple foods in Asia and Africa, are recently receiving a growing interest in Western countries. The object of this work is the characterization of a fermented wheat used as a food ingredient and dietary supplement. To this aim, the phenolic composition, the activity on protein tyrosine phosphatase 1B (PTP1B), an enzyme overexpressed in type-II diabetes, the in vitro prebiotic properties on Lactobacillus reuteri and the microbial composition were investigated. Basic and acidic hydrolysis were tested for an exhaustive recovery of bound phenols: the acidic hydrolysis gave best yields. Methyl ferulate and neocarlinoside were identified for the first time in wheat. The inhibitory power of the extracts of several batches were investigated on PTP1B enzyme. The product was not able to inhibit the enzyme, otherwise, for the first time, a complete inhibition was observed for schaftoside, a major C -flavonoid of wheat. The microbial composition was assessed identifying Lactobacillus, Enterococcus , and Pediococcus as the main bacterial species. The fermented wheat was a suitable substrate for the grown of L. reuteri , recognized for its health properties in the human gut. The proposed method for phenols is easier compared to those based on strong basic hydrolysis; our results assessed the bound phenols as the major fraction, differently from that suggested by the literature for fermented cereals.

The biological and functional heterogeneity between tumors-both across and within cancer types-poses a challenge for immunotherapy. To understand the factors underlying tumor immune heterogeneity and immunotherapy sensitivity, we established a library of congenic tumor cell clones from an autochthonous mouse model of pancreatic adenocarcinoma. These clones generated tumors that recapitulated T cell-inflamed and non-T-cell-inflamed tumor microenvironments upon implantation in immunocompetent mice, with distinct patterns of infiltration by immune cell subsets. Co-injecting tumor cell clones revealed the non-T-cell-inflamed phenotype is dominant and that both quantitative and qualitative features of intratumoral CD8 + T cells determine response to therapy. Transcriptomic and epigenetic analyses revealed tumor-cell-intrinsic production of the chemokine CXCL1 as a determinant of the non-T-cell-inflamed microenvironment, and ablation of CXCL1 promoted T cell infiltration and sensitivity to a combination immunotherapy regimen. Thus, tumor cell-intrinsic factors shape the tumor immune microenvironment and influence the outcome of immunotherapy., (Copyright © 2018. Published by Elsevier Inc.)

Epithelial-mesenchymal transition (EMT) is strongly implicated in tumor cell invasion and metastasis. EMT is thought to be regulated primarily at the transcriptional level through the repressive activity of EMT transcription factors. However, these classical mechanisms have been parsed out almost exclusively in vitro, leaving questions about the programs driving EMT in physiological contexts. Here, using a lineage-labeled mouse model of pancreatic ductal adenocarcinoma to study EMT in vivo, we found that most tumors lose their epithelial phenotype through an alternative program involving protein internalization rather than transcriptional repression, resulting in a "partial EMT" phenotype. Carcinoma cells utilizing this program migrate as clusters, contrasting with the single-cell migration pattern associated with traditionally defined EMT mechanisms. Moreover, many breast and colorectal cancer cell lines utilize this alternative program to undergo EMT. Collectively, these results suggest that carcinoma cells have different ways of losing their epithelial program, resulting in distinct modes of invasion and dissemination., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Comprehensive molecular analysis of rare circulating tumor cells (CTCs) and cell clusters is often hampered by low throughput and purity, as well as cell loss. To address this, we developed a fully integrated platform for flow cytometry-based isolation of CTCs and clusters from blood that can be combined with whole transcriptome analysis or targeted RNA transcript quantification. Downstream molecular signature can be linked to cell phenotype through index sorting. This newly developed platform utilizes in-line magnetic particle-based leukocyte depletion, and acoustic cell focusing and washing to achieve >98% reduction of blood cells and non-cellular debris, along with >1.5 log-fold enrichment of spiked tumor cells. We could also detect 1 spiked-in tumor cell in 1 million WBCs in 4/7 replicates. Importantly, the use of a large 200μm nozzle and low sheath pressure (3.5 psi) minimized shear forces, thereby maintaining cell viability and integrity while allowing for simultaneous recovery of single cells and clusters from blood. As proof of principle, we isolated and transcriptionally characterized 63 single CTCs from a genetically engineered pancreatic cancer mouse model (n = 12 mice) and, using index sorting, were able to identify distinct epithelial and mesenchymal sub-populations based on linked single cell protein and gene expression.