Molecular signatures have identified several subsets of diffuse large B cell lymphoma (DLBCL) and rational targets within the B cell receptor (BCR) signaling axis. The OxPhos-DLBCL subset, which ...harbors the signature of genes involved in mitochondrial metabolism, is insensitive to inhibition of BCR survival signaling but is functionally undefined. We show that, compared with BCR-DLBCLs, OxPhos-DLBCLs display enhanced mitochondrial energy transduction, greater incorporation of nutrient-derived carbons into the tricarboxylic acid cycle, and increased glutathione levels. Moreover, perturbation of the fatty acid oxidation program and glutathione synthesis proved selectively toxic to this tumor subset. Our analysis provides evidence for distinct metabolic fingerprints and associated survival mechanisms in DLBCL and may have therapeutic implications.
► Glucose and fatty acids are differentially utilized in molecular subsets of DLBCL ► Mitochondria are predominant in the metabolic signature of BCR-independent DLBCLs ► Fatty acid oxidation and glutathione synthesis are relevant to OxPhos-DLBCL survival ► Fuel utilization pathways define additional levels of heterogeneity in DLBCL
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•Eudragit® L100/chitosan blend forms bilayer films in 2-propanol after evaporation.•Higher Eudragit® L100 and plasticizer content improves the mechanical properties.•Mucoadhesion is ...not influenced by the amount of Eudragit® L100 or plasticizers.•The pH-responsive drug release is explained by the layer based on Eudragit® L100.•Higher Eudragit® L100 and plasticizer content allows a more controlled release.
The high rate of HIV new infections and AIDS-related deaths each year make prevention tools still necessary today. Different dosage forms – including films – for vaginal administration of antiretroviral drugs have been developed for this purpose. Six batches of Tenofovir-loaded films were formulated based on Eudragit® L100 (EL100) and chitosan, containing triethyl citrate and glycerol. In all the cases films structured in two layers – the upper layer mainly attributed to EL100 and the lower layer to chitosan – were revealed by SEM. A higher content in EL100 and plasticizers improves the mechanical properties and control over drug release in the vaginal medium without affecting mucoadhesion. The EL100-based layer acts as a structuring agent that controls Tenofovir release for days in the vaginal medium while it occurs in a few hours in the presence of seminal fluid. Bilayer films with the highest tested content of EL100 and plasticizers would be the most suitable as vaginal microbicides as they are easier to administer due to their excellent mechanical properties and they offer more comfortable posology and enhanced protection against HIV during intercourse due to their pH-responsive release of Tenofovir.
Salt content standardization in dry-cured ham production is complex as there are many factors that influence salt uptake. The aim of this work was to study and model salt uptake as a function of ...salting time and the characteristics of green ham in two different ham types using a large amount of data obtained from an inline non-invasive characterization technology. The usefulness of these models to standardize the salt content of the industrial production was also evaluated. Magnetic induction technology was used to characterize lean and fatty hams in order to further model the salt uptake using different statistical methods. Salt content increased with the salting time showing an asymptotic behaviour which was lower in fatty hams. The best models were those that considered the salting time, weight, and fat content of hams as model independent variables. These models are helpful for salt content standardization within the industry and can be used to simulate process modifications and avoid the time-consuming and costly trial and error tests.
•Salt content standardization of the dry-cured ham (DCH) production is complex.•Traditional DCH classification systems are not sufficient to reduce salt content variability.•Massive data obtained non-invasively shows salt uptake follows asymptotic behaviour.•Fatty hams showed lower salt uptake values than lean hams throughout salting.•Salt uptake predictive models can help to standardize the production salt content.
Changes in mitochondrial morphology are associated with nutrient utilization, but the precise causalities and the underlying mechanisms remain unknown. Here, using cellular models representing a wide ...variety of mitochondrial shapes, we show a strong linear correlation between mitochondrial fragmentation and increased fatty acid oxidation (FAO) rates. Forced mitochondrial elongation following MFN2 over‐expression or DRP1 depletion diminishes FAO, while forced fragmentation upon knockdown or knockout of MFN2 augments FAO as evident from respirometry and metabolic tracing. Remarkably, the genetic induction of fragmentation phenocopies distinct cell type‐specific biological functions of enhanced FAO. These include stimulation of gluconeogenesis in hepatocytes, induction of insulin secretion in islet β‐cells exposed to fatty acids, and survival of FAO‐dependent lymphoma subtypes. We find that fragmentation increases long‐chain but not short‐chain FAO, identifying carnitine O‐palmitoyltransferase 1 (CPT1) as the downstream effector of mitochondrial morphology in regulation of FAO. Mechanistically, we determined that fragmentation reduces malonyl‐CoA inhibition of CPT1, while elongation increases CPT1 sensitivity to malonyl‐CoA inhibition. Overall, these findings underscore a physiologic role for fragmentation as a mechanism whereby cellular fuel preference and FAO capacity are determined.
Synopsis
Metabolic fuel preference is associated with mitochondrial architecture, yet the mechanisms that connect shape and function remain unknown. This study demonstrates selective coupling between mitochondrial architecture and fatty acid (FA) utilization, uncovering an unexpected connection between mitochondrial fusion and fission, and fuel choice.
Long‐chain FA oxidation capacity increases upon mitochondrial fragmentation and decreases upon mitochondrial elongation.
Activity of CPT1, a rate‐limiting enzyme in fatty acid oxidation, is inhibited by the elongation of mitochondria, but enhanced by their fragmentation.
Mitochondrial fragmentation reduces CPT1's sensitivity to its endogenous inhibitor, malonyl‐CoA, which is a product of pyruvate metabolism and the first metabolite in FA synthesis.
Fragmentation‐induced FA utilization has cell type‐specific functional consequences, causing increased hepatic gluconeogenesis, induction of insulin secretion at non‐stimulatory glucose levels, and enhanced survival of OXPHOS‐dependent lymphoma cells.
Mitochondrial fragmentation increases beta‐oxidation and cellular lipid metabolism via de‐repressing CPT1.
Lymph node fibroblastic reticular cells (FRCs) respond to signals from activated T cells by releasing nitric oxide, which inhibits T cell proliferation and restricts the size of the expanding T cell ...pool. Whether interactions with FRCs also support the function or differentiation of activated CD8
T cells is not known. Here we report that encounters with FRCs enhanced cytokine production and remodeled chromatin accessibility in newly activated CD8
T cells via interleukin-6. These epigenetic changes facilitated metabolic reprogramming and amplified the activity of pro-survival pathways through differential transcription factor activity. Accordingly, FRC conditioning significantly enhanced the persistence of virus-specific CD8
T cells in vivo and augmented their differentiation into tissue-resident memory T cells. Our study demonstrates that FRCs play a role beyond restricting T cell expansion-they can also shape the fate and function of CD8
T cells.
Byproducts of arachidonic (AA) and docosahexaenoic acid (DHA) oxidation are highly relevant for the study of free radical associated conditions in the perinatal period. Plasma metabolites can provide ...the clinician with a snapshot of the oxidant status of patients before and after specific clinical interventions (e.g.: supplementation with oxygen). We describe a new andreliable ultra-performance liquid mass spectrometry method to determine F2-isoprostanes and other byproducts (isoprostanes, isofurans, neuroprostanes, neurofurans) in newborn serum samples. Cord blood samples were obtained from severely depressed newborn infants (Apgar score 1 min < 3; arterial cord pH < 7.00), and aliquoted for serum determination and stored at −80 °C. A UHPLC-MS/MS method was employed. It has a series of technical advantages: simple sample treatment; reduced sample volume (100 μL) which is essential for preterm neonates with low circulating blood volume, high throughput of sample analysis (96 samples in less than 24 h) and high selectivity for different isoprostanes isomers. Excellent sensitivity was achieved within limits of detection between 0.06 and 4.2 nmol L−1, which renders this method suitable to monitoranalyte concentration in newborn samples. The method's precision was satisfactory; with coefficients of variation around 5–12% (intra-day) and 7–17% (inter-day). The reliability of the described method was assessed by analysis of spiked serum samples obtaining recoveries between 70% and 120%. The proposed method has rendered suitable for serum determination for newborn babies at risk of oxygen free radical associated conditions.
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•A validated liquid chromatographic method to determine peroxidation lipid biomarkers.•Serum samples from severely depressed newborn infants (Apgar score 1 min <3; arterial cord pH <7.00).•Small sample volume and simple sample treatment.•High throughput of sample analysis and high selectivity for different isoprostanes isomers.
The end of the elaboration process of dry-cured ham is currently decided by product weight loss and/or by an expert who carries out an evaluation of the tactile texture on the surface. The objective ...of this study was to define the optimal measurement conditions of an instrumental texture analysis on the surface of the dry-cured ham (ITAS), to define the end of process. 120 dry-cured hams were classified by experts into Hard (appropriate) or Soft (non-appropriate) texture groups and used to perform compression tests using different probes on three anatomical positions. Results showed that the small probe in position 2 gave the most discriminant conditions, providing representative information of the internal texture. Although classification using only weight loss was possible with an accuracy rate of 80.4% or 66.7% depending on the weight loss, the maximum classification accuracy was obtained when using ITAS in combination with weight loss. Further studies at industrial level are needed.
•To define dry-cured ham (DCH) end of process according to its texture is a challenge.•A new Instrumental Texture Analysis on the surface of the ham (ITAS) is developed.•ITAS using a small probe in position 2 can identify finished DCH with low error.•ITAS in combination with weight loss improves classification ability.
Diffuse large B-cell lymphomas (DLBCLs) are a highly heterogeneous group of tumors in which subsets share molecular features revealed by gene expression profiles and metabolic fingerprints. While ...B-cell receptor (BCR)-dependent DLBCLs are glycolytic, OxPhos-DLBCLs rely on mitochondrial energy transduction and nutrient utilization pathways that provide pro-survival benefits independent of BCR signaling. Integral to these metabolic distinctions is elevated mitochondrial electron transport chain (ETC) activity in OxPhos-DLBCLs compared with BCR-DLBCLs, which is linked to greater protein abundance of ETC components. To gain insights into molecular determinants of the selective increase in ETC activity and dependence on mitochondrial energy metabolism in OxPhos-DLBCLs, we examined the mitochondrial translation pathway in charge of the synthesis of mitochondrial DNA encoded ETC subunits. Quantitative mass spectrometry identified increased expression of mitochondrial translation factors in OxPhos-DLBCL as compared with the BCR subtype. Biochemical and functional assays indicate that the mitochondrial translation pathway is required for increased ETC activity and mitochondrial energy reserves in OxPhos-DLBCL. Importantly, molecular depletion of several mitochondrial translation proteins using RNA interference or pharmacological perturbation of the mitochondrial translation pathway with the FDA-approved inhibitor tigecycline (Tigecyl) is selectively toxic to OxPhos-DLBCL cell lines and primary tumors. These findings provide additional molecular insights into the metabolic characteristics of OxPhos-DLBCLs, and mark the mitochondrial translation pathway as a potential therapeutic target in these tumors.
Highlights • Cells can use different metabolic fuels as an adaptive advantage to stress. • Proliferation and differentiation are accompanied by metabolic rewiring. • Mitochondrial architecture and ...metabolic compartmentalization influence fuel choice.
Protective coatings that retard ionic diffusion at high temperature in the presence of steam are of interest for the energy industry. A CrAlSiN monolayer and two CrN/AlSiN coatings of periodic layers ...of different thicknesses deposited on P92 steel by cathodic arc evaporation (CAE) were studied. Their protection capabilities under the working conditions of ultrasupercritical steam turbines (at 650 °C in 100% steam atmosphere) up to 2000 h were assessed. Several characterization techniques were used to achieve this objective, such as gravimetric analysis, X-ray diffraction (XRD), scanning electron microscopy with an energy dispersive detector (SEM-EDS), field emission scanning electron microscopy (FE-SEM) and glow discharge optical emission spectrometry (GDOES).
After 2000 h, the oxidation resistance of the coated samples showed a significant improvement compared to that of the bare substrate. This behavior is explained by the formation of a protective outer layer of chromia (Cr2O3), which prevents or reduces the inward and outward diffusion of oxygen and metallic cations, respectively, leading to a reduction of the oxidation rate at working conditions. Oxidation kinetics of the coated samples follows a parabolic trend at the first stages of oxidation, according with the development of a protective oxide on the surface during the thermal treatment. However, after a certain time an apparent change in the kinetics of the oxide growth is observed, that has been related to the detachment of the oxidized macrodroplets of the evaporated materials, which are characteristic of the CAE technique. Nevertheless, this phenomenon does not decrease the oxidation resistance of the coatings.
•CrAlSiN based coatings deposited on P92 steel by cathodic arc evaporation.•Composition of coatings ranged 3–4 at.% Si with the Cr/Al ratio at ∼4.5.•Coating and substrate oxidation resistances were compared at 650 °C in 100% steam.•After 2000 h, Cr2O3 forms at the surface and CrN partially transforms to Cr2N.•Benefits of multilayer architecture are highlighted with respect to the monolayer.
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