Lipotoxicity is a form of cellular stress caused by the accumulation of lipids resulting in mitochondrial dysfunction and insulin resistance in muscle. Previously, we demonstrated that the mitophagy ...receptor BNIP3L/Nix is responsive to lipotoxicity and accumulates in response to a high-fat (HF) feeding. To provide a better understanding of this observation, we undertook gene expression array and shot-gun metabolomics studies in soleus muscle from rodents on an HF diet. Interestingly, we observed a modest reduction in several autophagy-related genes. Moreover, we observed alterations in the fatty acyl composition of cardiolipins and phosphatidic acids. Given the reported roles of these phospholipids and BNIP3L in mitochondrial dynamics, we investigated aberrant mitochondrial turnover as a mechanism of impaired myocyte insulin signaling. In a series of gain-of-function and loss-of-function experiments in rodent and human myotubes, we demonstrate that BNIP3L accumulation triggers mitochondrial depolarization, calcium-dependent activation of DNM1L/DRP1, and mitophagy. In addition, BNIP3L can inhibit insulin signaling through activation of MTOR-RPS6KB/p70S6 kinase inhibition of IRS1, which is contingent on phosphatidic acids and RHEB. Finally, we demonstrate that BNIP3L-induced mitophagy and impaired glucose uptake can be reversed by direct phosphorylation of BNIP3L by PRKA/PKA, leading to the translocation of BNIP3L from the mitochondria and sarcoplasmic reticulum to the cytosol. These findings provide insight into the role of BNIP3L, mitochondrial turnover, and impaired myocyte insulin signaling during an overfed state when overall autophagy-related gene expression is reduced. Furthermore, our data suggest a mechanism by which exercise or pharmacological activation of PRKA may overcome myocyte insulin resistance.
Abbreviations: BCL2: B cell leukemia/lymphoma 2; BNIP3L/Nix: BCL2/adenovirus E1B interacting protein 3-like; DNM1L/DRP1: dynamin 1-like; FUNDC1: FUN14 domain containing 1; IRS1: insulin receptor substrate 1; MAP1LC3A/LC3: microtubule-associated protein 1 light chain 3 alpha; MFN1: mitofusin 1; MFN2: mitofusin 2; MTOR: mechanistic target of rapamycin kinase; OPA1: OPA1 mitochondrial dynamin like GTPase; PDE4i: phosphodiesterase 4 inhibitor; PLD1: phospholipase D1; PLD6: phospholipase D family member 6; PRKA/PKA: protein kinase, AMP-activated; PRKCD/PKCδ: protein kinase C, delta; PRKCQ/PKCθ: protein kinase C, theta; RHEB: Ras homolog enriched in brain; RPS6KB/p70S6K: ribosomal protein S6 kinase; SQSTM1/p62: sequestosome 1; YWHAB/14-3-3β: tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein beta
Pulses are known to be nutritious foods but are susceptible to oxidation due to the reaction of lipoxygenase (LOX) with linolenic and linoleic acids which can lead to off flavors caused by the ...formation of volatile organic compounds (VOCs). Infrared micronization at 130 and 150 °C was investigated as a heat treatment to determine its effect on LOX activity and VOCs of chickpea and green lentil flour. The pulse flours were added to low‐fat beef burgers at 6% and measured for consumer acceptability and physicochemical properties. Micronization at 130 °C significantly decreased LOX activity for both flours. The lentil flour micronized at 150 °C showed a further significant decrease in LOX activity similar to that of the chickpea flour at 150 °C. The lowering of VOCs was accomplished more successfully with micronization at 130 °C for chickpea flour while micronization at 150 °C for the green lentil flour was more effective. Micronization minimally affected the characteristic fatty acid content in each flour but significantly increased omega‐3 and n‐6 fatty acids at 150 °C in burgers with lentil and chickpea flours, respectively. Burgers with green lentil flour micronized at 130 and 150 °C, and chickpea flour micronized at 150 °C were positively associated with acceptability. Micronization did not affect the shear force and cooking losses of the burgers made with both flours. Formulation of low‐fat beef burgers containing 6% micronized gluten‐free binder made from lentil and chickpea flour is possible based on favorable results for physicochemical properties and consumer acceptability.
Alpha linolenic acid (ALA) is an 18-carbon essential fatty acid found in plant-based foods and oils. While much attention has been placed on conversion of ALA to long chain polyunsaturated fatty ...acids, alternative routes of ALA metabolism exist and may lead to formation of other bioactive metabolites of ALA. The current study employed a non-targeted metabolomics approach to profile ALA metabolites that are significantly upregulated by ALA treatment. H4IIE hepatoma cells (n=3 samples per time point) were treated with 60 μM ALA or vehicle for 0, 0.25, 0.5, 1, 2, 3, 4, 6, 8, and 12 h. Samples were then extracted with methanol and analyzed using high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry. We observed selective changes in ALA incorporation into phospholipid classes and subclasses over the 12 h following ALA treatment. While levels of specific molecular species of ALA-containing phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, and lysophospholipids were elevated with ALA treatment, others were not affected. Of the phospholipids that were increased, some (e.g., PC18:3/18:1, PC18:3/18:4, PE18:3/18:2, PE18:3/18:3) were elevated almost immediately after exposure to ALA, while others e.g., PE18:1/18:3 PA18:3/22:6, and PA18:3/18:2) were not elevated until several hours after ALA treatment. Overall, these results suggest that ALA incorporation into phospholipids is selective and support a metabolic hierarchy for ALA incorporation into specific phospholipids. Given the functionality of phospholipids based on their fatty acid composition, future studies will need to investigate the implications of ALA incorporation into specific phospholipids on cell function.
OBJECTIVES: Lung cancer is one of the most lethal cancers. Currently, there are no biomarkers for early detection, monitoring treatment response, and detecting recurrent lung cancer. We undertook ...this study to determine if 1^H magnetic resonance spectroscopy (MRS) of sputum and exhaled breath condensate (EBC), as a noninvasive tool, can identify metabolic biomarkers of lung cancer. MATERIALS AND METHODS: Sputum and EBC samples were collected from 20 patients, comprising patients with pathologically confirmed nonsmall cell lung cancer (n = 10) and patients with benign respiratory conditions (n = 10). Both sputum and EBC samples were collected from 18 patients; 2 patients provided EBC samples only. ^1H MR spectra were obtained on a Bruker Avance 400 MHz nuclear magnetic resonance (NMR) spectrometer. Sputum samples were further confirmed cytologically to distinguish between true sputum and saliva. RESULTS: In the EBC samples, median concentrations of propionate, ethanol, acetate, and acetone were higher in lung cancer patients compared to the patients with benign conditions. Median concentration of methanol was lower in lung cancer patients (0.028 mM) than in patients with benign conditions (0.067 mM; P = 0.028). In the combined sputum and saliva and the cytologically confirmed sputum samples, median concentrations of N-acetyl sugars, glycoprotein, propionate, lysine, acetate, and formate were lower in the lung cancer patients than in patients with benign conditions. Glucose was found to be consistently absent in the combined sputum and saliva samples (88%) as well as in the cytologically confirmed sputum samples (86%) of lung cancer patients. CONCLUSION: Absence of glucose in sputum and lower concentrations of methanol in EBC of lung cancer patients discerned by ^1H MRS may serve as metabolic biomarkers of lung cancer for early detection, monitoring treatment response, and detecting recurrence.
Abstract
The PI3K signalling pathway is known to regulate B cell metabolic programming upon activation, but the mechanisms involved are not well understood. Here we find that the PI3K pathway ...controls reprogramming of hexokinases (HKs), the enzymes that convert glucose into glucose-6-phosphate as a key rate-limiting step of glycolysis and other metabolic pathways. In primary mouse B cells, PI3K pathway inhibition substantially impaired the activation-induced increase in extracellular acidification rates (ECAR), a measure of glycolysis. In contrast, B cells isolated from PI3Kdelta gain-of-function mutant mice exhibit elevated ECAR. We find that B cell activation substantially elevates protein levels of HK2 and HK3 isoforms, but not HK1, in a PI3K-dependant manner. PI3K or mTOR inhibition significantly reduced induction of HK2 and HK3 expression, whereas Akt inhibition did not affect HK isoform expression. In human B lymphoma cells, HK isoforms differ significantly in their degree of mitochondrial localization, with HK1 being mitochondrial, HK3 being cytoplasmic and HK2 present in both mitochondria and cytoplasm. To assess whether HK isoforms have unique non-redundant functions, HK2-deficient B lymphoma cells were generated and were found to exhibit decreased ECAR as well as significant changes in metabolomic profile, despite normal expression and localization of HK1 and HK3. Taken together, our study reveals that PI3K-dependant reprogramming of hexokinase isoforms can impact on B cell glycolysis and other metabolic pathways. Studies in progress are examining the functional importance of HK2 in antibody responses using mice with B cell-specific deletion of this enzyme.
Although choline is essential for brain development and neural function, the effect of choline on retina function is not well understood. This study examined the effects of choline on neural tissues ...of brain and retina, and membrane phospholipid (PL) composition during fetal development.
Pregnant C57BL/6 mice were fed one of 4 choline modified diets: i) control (Cont, 2.5g/kg), ii) choline deficient (Def, 0g/kg), iii) supplemented with choline chloride (Cho, 10g/kg) and iv) supplemented with egg phosphatidylcholine (PC, 10g/kg). At postnatal day (PD) 21, pups were weaned onto their mothers' respective diets until PD 45. Spatial memory was measured using the Morris Water Maze; retina function by electroretinogram (ERG); and PL composition with nuclear magnetic resonance spectroscopy.
Cho and PC supplementation enhanced cued learning and spatial memory abilities, respectively (p Def > PC > Cho, with no statistically significant alterations in cone-driven responses. There were no differences in the composition of major PLs in the brain and retina. In the brain, subclasses of ether PL, alkyl acyl- phosphatidylethanolamine (PEaa) and phosphatidylcholine (PCaa) were significantly greater among the PC supplemented group in comparison to the Def group.
These results indicate that while choline supplementation during gestation to an early developmental period is beneficial for spatial memory, contributions to retina function are minor. Assessment with a larger sample size of retinas could warrant the essentiality of choline for retina development.
Fetal alcohol spectrum disorder (FASD) is a major public health issue that encompass an array of physical, neurological, and behavioral effects due to alcohol consumption during pregnancy. The ...classical biomarkers of FASD that are currently used lack sensitivity and specificity, and as such there is an opportunity through the use of novel metabolomics analysis to identify new biomarkers to identify those at risk for FASD, which could more effectively aid in early intervention. The focus of this minireview is to identify current work that is being done in the field of metabolomics in FASD in utero, and to highlight promising metabolites that could act as biomarkers in the future. We will conclude with suggestions for further research, as there is a large gap of knowledge in this particular area of metabolomics.
Development of bakery products containing milled flaxseed, could have potential health benefits due to its high antioxidant, lignan and omega-3 fatty acid alpha linolenic acid content. However the ...effect of adding flaxseed, especially in the high amounts required for use as the food supplement in clinical trials, may affect the flavor characteristics. The objective of this study was to evaluate the flavor profile of different muffin and snack bar formulations with and without milled flaxseed. Three snack bar formulations and two muffin formulations with and without flaxseed were evaluated by ten and eleven trained assessors respectively. The intensity of seven aroma and eight flavor attributes for snack bars, and nine aroma and ten flavor attributes for muffins were measured on 15-cm line scales. Both the orange cranberry and apple spice muffin formulations with flaxseed had significantly lower sweetness and vanilla aroma and flavor intensities, and significantly higher grain/flax aroma and flavor, and bitter taste compared to the non-flax muffin. Of the two flavorings apple spice appeared to have the best potential, since the apple aroma and flavor were significantly higher for the flax muffin compared to the non-flax muffin. The orange cranberry, gingerbread raisin and cappuccino chocolate chip snack bar formulations with and without flaxseed showed no significant differences in intensities for the grain/flax aroma and flavor. The gingerbread raisin snack bar formulations showed no significant differences for any other attributes except for higher spice aroma in the flax sample, which makes it a promising flavoring option.
Yellow pea (Pisumsativum L.) is an economically rich source of nutrients with health‐promoting effects. However, the consumption of pea ingredients is minimal due to their off‐flavor characteristics. ...The present study investigated the effect of Revtech heat treatment on the chemical profile and volatile compounds in split yellow pea flour. Revtech treatment (RT) was applied at 140°C with a residence time of 4 min in dry condition (RT 0%) and in the presence of 10% steam (RT 10%). Both thermal treatments resulted in a significant reduction (p < 0.05) in lipoxygenase activity and the concentration of key beany‐related odors such as heptanal, (E)‐2‐heptenal, 1‐octen‐3‐ol, octanal, and (E)‐2‐octenal. In addition, RT 10% resulted in a significant reduction in pentanal, 1‐penten‐3‐ol, hexanal, and 1‐hexanol compared to untreated flour. The content of known precursors of lipoxygenase such as linoleic and linolenic acids was found in higher concentrations in heat‐treated flours, indicating the efficacy of Revtech technology in minimizing the degradation of polyunsaturated fatty acids. No significant changes in the amino acid composition or the 29 selected phenolic compounds in pea flours were observed with Revtech processing except for two compounds, caffeic acid and gallocatechin, which were found at higher concentrations in RT 0%.
Practical Application
Thermal processing of split yellow pea flours at 140°C using Revtech technology successfully decreased the concentrations of volatile compounds responsible for beany off‐flavor while improving the nutritional quality of studied yellow pea flours. These results provide valuable information to the food industry for developing novel pulse‐based products with enhanced sensory characteristics.
PDF
