AMPK regulates phagophore-to-autophagosome maturation.

Autophagy is an important metabolic pathway that can non-selectively recycle cellular material or lead to targeted degradation of protein aggregates or damaged organelles. Autophagosome formation starts with autophagy factors accumulating on lipid vesicles containing ATG9. These phagophores attach to donor membranes, expand via ATG2-mediated lipid transfer, capture cargo, and mature into autophagosomes, ultimately fusing with lysosomes for their degradation.

FooDOxS: a database of oxidized sterols content in foods.

Dietary oxidized sterols (DOxS) are cholesterol-like molecules known to exert pro-inflammatory, pro-oxidant, and pro-apoptotic effects, among others. We present the FooDOxS database, a comprehensive compilation of DOxS content in over 1680 food items from 120 publications across 25 countries, augmented by data generated by our group. This database reports DOxS content in foods classified under the NOVA and What We Eat in America (WWEIA) systems, allowing a comprehensive and statistically robust summary of DOxS content in foods.

Oxidative Status of Ultra-Processed Foods in the Western Diet.

Ultra-processed foods (UPFs) have gained substantial attention in the scientific community due to their surging consumption and potential health repercussions. In addition to their well-established poor nutritional profile, UPFs have been implicated in containing various dietary oxidized sterols (DOxSs). These DOxSs are associated with a spectrum of chronic diseases, including cardiometabolic conditions, cancer, diabetes, Parkinson's, and Alzheimer's disease.

Cholesterol crystals induce mechanical trauma, inflammation, and neo-vascularization in solid cancers as in atherosclerosis.

Background And Aims: Cancer and atherosclerosis share common risk factors and inflammatory pathways that promote their proliferation via vascular endothelial growth factor (VEGF). Because CCs cause mechanical injury and inflammation in atherosclerosis, we investigated their presence in solid cancers and their activation of IL-1β, VEGF, CD44, and Ubiquityl-Histone H2B (Ub-H2B), that promote cancer growth.

Methods: Tumor specimens from eleven different types of human cancers and atherosclerotic plaques were assessed for CCs, free cholesterol content and IL1-β by microscopy, immunohistochemistry, and biochemical analysis.

AMPK Regulates Phagophore-to-Autophagosome Maturation.

Autophagy is an important metabolic pathway that can non-selectively recycle cellular material or lead to targeted degradation of protein aggregates or damaged organelles. Autophagosome formation starts with autophagy factors accumulating on lipid vesicles containing ATG9. These phagophores attach to donor membranes, expand via ATG2-mediated lipid transfer, capture cargo, and mature into autophagosomes, ultimately fusing with lysosomes for their degradation.

Tracking the transition from an ATG9A vesicle to an autophagosome.

ATG: autophagy-related proteins; ULK1/2: Unc-51-Like activating Kinases; PI3Ks: Phosphoinositide 3-Kinases; ATG2A: autophagy-related protein 2A; ATG5: autophagy-related protein 5; ATG16: autophagy-related protein 16; ATG8: autophagy-related protein 8; U2OS: human bone osteosarcoma epithelial cell; LC3B: microtubule-associated protein 1A/1B Light Chain 3B; GABARAPL1: GABA type A Receptor-Associated Protein Like 1; ATG9A: autophagy-related protein 9A; ATG13: autophagy-related protein 13; SQSTM1: Sequestosome-1/p62; WIPI2: WD repeat domain, Phosphoinositide Interacting 2; PI3P: Phosphoinositide-3-phosphate.

Quantitative analysis of autophagy reveals the role of ATG9 and ATG2 in autophagosome formation.

Autophagy is a catabolic pathway required for the recycling of cytoplasmic materials. To define the mechanisms underlying autophagy it is critical to quantitatively characterize the dynamic behavior of autophagy factors in living cells. Using a panel of cell lines expressing HaloTagged autophagy factors from their endogenous loci, we analyzed the abundance, single-molecule dynamics, and autophagosome association kinetics of autophagy proteins involved in autophagosome biogenesis.

Dietary exposure assessment of infant formula and baby foods' oxidized lipids in the US population.

Baby Foods (BFs) and Infant formulas (IFs) are the main sources of nutrition for an infant throughout the 1st year of life. Various enriched products are commercially available for parents seeking to fulfill their baby's nutritional needs. Consequently, different bioactive lipids are present in BFs and IFs, including dietary oxysterols (DOxS), whose known toxicity has been associated with mutagenicity, cancer, and other chronic diseases.

Evaluation of the nutritional quality of ultra-processed foods (ready to eat + fast food): Fatty acids, sugar, and sodium.

The average American consumes more than 50% of their total dietary energy from ultra-processed foods (UPFs). From a nutritional standpoint, as UPFs intake increases, fiber, vitamin, and mineral intake decrease. High consumption of UPFs, mainly from fast foods (FF) and ready-to-eat (RTE) food items, emerges as a critical public health concern linking nutritional quality and food safety.

Alkaloid profiling of Italian alpine herbs using high resolution mass spectrometry (Orbitrap-MS).

Alpine pastures have not yet been extensively studied with regard to the presence of alkaloids in herbaceous plants. In this work the alkaloid profiles were characterised from a selection of 62 herbs collected from alpine pastures in north-eastern Italy. High-performance liquid chromatography coupled to a hybrid quadrupole-orbitrap mass spectrometer was used to evaluate the presence of 41 different alkaloids and quantify them using a targeted approach.

Lipid profiling and dietary assessment of infant formulas reveal high intakes of major cholesterol oxidative product (7-ketocholesterol).

Approximately two-thirds of US infants receive infant formula (IF) as a primary or sole nutritional source during the first six months of life. IF is available in a variety of commercial presentations; from a manufacturing standpoint, they can be categorized as powder- (PIF) or liquid- (LIF) based formulations. Thirty commercial IFs were analyzed in their oxidative and non-oxidative lipid profiles.

Efficacy and Safety of Non-Anesthesiologist Administration of Propofol Sedation in Endoscopic Ultrasound: A Propensity Score Analysis.

In spite of promising preliminary results, evidence supporting the use of non-anesthesiologist-administered propofol sedation (NAAP) in endoscopic ultrasound (EUS) procedures is still limited. The aim of this manuscript was to examine the safety and efficacy of NAAP as compared to anesthesiologist-administered propofol sedation in EUS procedures performed in a referral center. Out of 832 patients referred to our center between 2016 and 2019, after propensity score matching two groups were compared: 305 treated with NAAP and 305 controls who underwent anesthesiologist-administered propofol sedation.

Flavonoids Ability to Disrupt Inflammation Mediated by Lipid and Cholesterol Oxidation.

Flavonoids are plant secondary metabolites that act as protectants against harmful effects of UV-B radiation inasmuch as biotic stress, conferring at the same time pigmentation of fruits and leaves [67]. The term "flavonoid" refers to phenolics having a basic skeleton of diphenylpropane (C6-C3-C6), which consists of two aromatic rings linked through three carbons that usually form an oxygenated heterocycle [25, 52]. Flavonoids are broken down into several different sub-categories based on their chemical structure.

Picturing the Membrane-assisted Choreography of Cytochrome P450 with Lipid Nanodiscs.

Cytochrome P450, a family of monooxygenase enzymes, is organized as a catalytic metabolon, and requires enzymatic partners as well as environmental factors that tune its complex dynamic activity. P450 and its reducing counterparts are membrane-bound proteins which are believed to dynamically interact to form functional complexes. Increasing experimental evidence signifies the role (s) of protein-lipid interactions in P450's catalytic function and efficiency.

The role of cholesterol oxidation products in food toxicity.

Food consumption can lead to the accumulation of certain chemical compounds able to exert toxic activities against humans. Of mayor interests are those molecules generated during food processing and handling, since their occurrence and distribution depend of many intrinsic and extrinsic factors. Cholesterol - a lipid constituent of mammalian cells - is the precursor of several toxic molecules known as cholesterol oxidation products (COPs).

Glycosylated simple phenolic profiling of food tannins using high resolution mass spectrometry (Q-Orbitrap).

Tannins are polyphenolic compounds extensively present in plants and used by food industry as processing aids. Due to the heterogeneity of plant sources, actions involved in food processing and tannin commercial costs can be different. In the last years different approaches aimed at correctly identifying the tannin botanical origin have been developed, in order to satisfy the industry's request to verify product labels.

Lipid-exchange in nanodiscs discloses membrane boundaries of cytochrome-P450 reductase.

Lipids are critical for the function of membrane proteins. NADPH-cytochrome-P450-reductase, the sole electron transferase for microsomal oxygenases, possesses a conformational dynamics entwined with its topology. Here, we use peptide-nanodiscs to unveil cytochrome-P450-reductase's lipid boundaries, demonstrating a protein-driven enrichment of ethanolamine lipids (by 25%) which ameliorates by 3-fold CPR's electron-transfer ability.

Non-targeted glycosidic profiling of international wines using neutral loss-high resolution mass spectrometry.

Many metabolites naturally occur as glycosides, since sugar moieties can be crucial for their biological activity and increase their water solubility. In the plant kingdom they may occur as glycosides or sugar esters, depending on precursor chemical structure, and in wine they have traditionally attracted attention due to their organoleptic properties, such as astringency and bitterness, and because they affect the colour and aroma of wines. A new approach directed at detailed description of glycosides in a large selection of monovarietal wines (8 samples each of Pinot Blanc, Muller Thurgau, Riesling, Traminer, Merlot, Pinot Noir and Cabernet Sauvignon) was developed by combining high performance liquid chromatography with high resolution tandem mass spectrometry.

Cytochrome-P450-Induced Ordering of Microsomal Membranes Modulates Affinity for Drugs.

Although membrane environment is known to boost drug metabolism by mammalian cytochrome P450s, the factors that stabilize the structural folding and enhance protein function are unclear. In this study, we use peptide-based lipid nanodiscs to "trap" the lipid boundaries of microsomal cytochrome P450 2B4. We report the first evidence that CYP2B4 is able to induce the formation of raft domains in a biomimetic compound of the endoplasmic reticulum.

Dissociation Constants of Cytochrome P450 2C9/Cytochrome P450 Reductase Complexes in a Lipid Bilayer Membrane Depend on NADPH: A Single-Protein Tracking Study.

Cytochrome P450-reductase (CPR) is a versatile NADPH-dependent electron donor located in the cytoplasmic side of the endoplasmic reticulum. It is an electron transferase that is able to deliver electrons to a variety of membrane-bound oxidative partners, including the drug-metabolizing enzymes of the cytochrome P450s (P450). CPR is also stoichiometrically limited compared to its oxidative counterparts, and hypotheses have arisen about possible models that can overcome the stoichiometric imbalance, including quaternary organization of P450 and diffusion-limited models.

Membrane environment drives cytochrome P450's spin transition and its interaction with cytochrome b.

Heme's spin-multiplicity is key in determining the enzymatic function of cytochrome P450 (cytP450). The origin of the low-spin state in ferric P450 is still under debate. Here, we report the first experimental demonstration of P450's membrane interaction altering its spin equilibrium which is accompanied by a stronger affinity for cytochrome b.

The effects of Cissampelos pareira extract on envenomation induced by Bothropsdiporus snake venom.

Ethnopharmacological Relevance: Ophidian accidents are a serious public health problem in Argentina; the Bothrops species is responsible for 97% of these accidents, and in particular, B. diporus is responsible for 80% of them. In the northeast of the country (Corrientes Provinces), Cissampelos pareira L.

Kinetic and Structural Characterization of the Effects of Membrane on the Complex of Cytochrome b and Cytochrome c.

Cytochrome b (cytb ) is a membrane protein vital for the regulation of cytochrome P450 (cytP450) metabolism and is capable of electron transfer to many redox partners. Here, using cyt c as a surrogate for cytP450, we report the effect of membrane on the interaction between full-length cytb and cyt c for the first time. As shown through stopped-flow kinetic experiments, electron transfer capable cytb - cyt c complexes were formed in the presence of bicelles and nanodiscs.

Fast analysis of quaternary ammonium pesticides in food and beverages using cation-exchange chromatography coupled with isotope-dilution high-resolution mass spectrometry.

A fast separation based on cation-exchange liquid chromatography coupled with high-resolution mass spectrometry is proposed for simultaneous determination of chlormequat, difenzoquat, diquat, mepiquat and paraquat in several food and beverage commodities. Solid samples were extracted using a mixture of water/methanol/formic acid (69.6:30:0.

Alkaloid profiling of herbal drugs using high resolution mass spectrometry.

Herbal infusions are consumed worldwide thanks to their "natural" beneficial effects, also due to the presence of alkaloids, although these compounds can have poisonous effects. A method combining online solid-phase purification with high resolution mass spectrometry was used to define the alkaloid profiles of 117 herbs and 7 commercial blends. Forty-one alkaloids were quantified in reference to analytical standards, while the presence of a further 116 was confirmed based on accurate mass, retention time, and fragmentation profile.