Lipid Utilization Research Articles

Impact of methimazole-induced hypothyroidism on postnatal swine.

Thyroid hormones regulate metabolic rate, nutrient utilization, growth, and development. Swine are susceptible to thyroid suppression in response to disease or environmental conditions, but the physiological impact of such disruption has not been established. The objective of this study was to evaluate the impact of hypothyroidism induced with the antithyroid medication methimazole (MMI). 10 mg/kg MMI significantly decreased circulating triiodothyronine (T3) for the duration of treatment but had only a transient effect on circulating thyroxine (T4). Thyroid tissue weight was significantly increased by more than 3.5-fold in response to MMI treatment. Histologically, the eosinophilic colloid was largely absent from the thyroid follicle which displayed a disorganized columnar epithelium consistent with goiter. MMI induced hypothyroidism has no effect on growth rate over 28 days. Hepatic expression of genes associated with thyroid metabolism (DIO1, DIO2, and DIO3), lipid utilization (CD36, FASN, and ACACA), apoptosis (TP53, PERP, SIVA1, and SFN) and proliferation (CDK1, CDK2, CDK4, and CDKN1A) were unaffected by treatment. Collectively these results demonstrate that MMI induces mild systemic hypothyroidism and pronounced goiter, indicating a strong homeostatic central regulation within the hypothalamic pituitary thyroid axis. This combined with limited peripheral effects, indicates resilience to hypothyroidism in modern swine.

Kombucha Tea-associated microbes remodel host metabolic pathways to suppress lipid accumulation.

The popularity of the ancient, probiotic-rich beverage Kombucha Tea (KT) has surged in part due to its purported health benefits, which include protection against metabolic diseases; however, these claims have not been rigorously tested and the mechanisms underlying host response to the probiotics in KT are unknown. Here, we establish a reproducible method to maintain C. elegans on a diet exclusively consisting of Kombucha Tea-associated microbes (KTM), which mirrors the microbial community found in the fermenting culture. KT microbes robustly colonize the gut of KTM-fed animals and confer normal development and fecundity. Intriguingly, animals consuming KTMs display a marked reduction in total lipid stores and lipid droplet size. We find that the reduced fat accumulation phenotype is not due to impaired nutrient absorption, but rather it is sustained by a programed metabolic response in the intestine of the host. KTM consumption triggers widespread transcriptional changes within core lipid metabolism pathways, including upregulation of a suite of lysosomal lipase genes that are induced during lipophagy. The elevated lysosomal lipase activity, coupled with a decrease in lipid droplet biogenesis, is partially required for the reduction in host lipid content. We propose that KTM consumption stimulates a fasting-like response in the C. elegans intestine by rewiring transcriptional programs to promote lipid utilization. Our results provide mechanistic insight into how the probiotics in Kombucha Tea reshape host metabolism and how this popular beverage may impact human metabolism.

Abstract 4448: Effects of adagrasib on cholesterol, lipid and glucose gene expression regulation in tumor xenograft models and patient samples

Abstract Introduction: Clinically active KRASG12C inhibitors have represented a key research breakthrough and led to novel treatment options for lung, colorectal and other cancer patients harboring this mutation. While treatment with KRASG12C inhibitors are clinically active in the majority of patients, the depth and duration of response is variable and most patients ultimately progress. This variation in therapeutic response highlights the need for a better understanding of drug mechanism of action and the identification of rational combination strategies. Oncogenic KRAS mutations hyperactivate the MAPK pathway, create an immunosuppressive tumor microenvironment, and promote glycolysis even in the presence of oxygen commonly known as the Warburg effect. This altered metabolic state is marked by increased glucose uptake and lactate production, providing necessary substrates for rapid tumor growth. Results: Pharmacological inhibition of KRASG12C with a potent and selective KRASG12C inhibitor, adagrasib, revealed marked alterations of metabolic gene expression programs in tumor samples collected from mouse xenograft studies and repeat biopsies provided by patients enrolled on adagrasib clinical trials. Upregulation of genes involved in reverse cholesterol transport including ApoE, NR1H3 (LXRa) and ABCA1 was observed across both preclinical tumor xenografts and patient samples following adagrasib treatment. Additionally, decreased expression of Glut1 and low-density lipoprotein receptor (LDLR) genes were also observed in both tumor models and patients suggesting that adagrasib treatment results in an extensive shift in uptake and utilization of cholesterol, lipid, and glucose. The impact of adagrasib treatment on circulating metabolic parameters in tumor xenograft-bearing mice were investigated in follow up studies. Conclusions: Taken together, these data suggest adagrasib modulates cholesterol efflux, glucose and lipid pathways and alters tumor and systemic metabolic regulation. Additional research may yield a targetable collateral vulnerability and rational combinatorial strategy. Citation Format: Fabiola Shelton, Natalie Hoffman, David Trinh, Xousaen M. Helu, Andrew Calinisan, Allan Hebbert, Larry Yan, David M. Briere, Laura Hover, Julio Fernandez-Benet, Kenna Anderes, James G. Christensen, Jill Hallin, Peter Olson. Effects of adagrasib on cholesterol, lipid and glucose gene expression regulation in tumor xenograft models and patient samples [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4448.

Acetyl-CoA carboxylase obstructs CD8+ T cell lipid utilization in the tumor microenvironment

The solid tumor microenvironment (TME) imprints a compromised metabolic state in tumor-infiltrating Tcells (TILs), hallmarked by the inability to maintain effective energy synthesis for antitumor function and survival. Tcells in the TME must catabolize lipids via mitochondrial fatty acid oxidation (FAO) to supply energy in nutrient stress, and it is established that Tcells enriched in FAO are adept at cancer control. However, endogenous TILs and unmodified cellular therapy products fail to sustain bioenergetics in tumors. We reveal that the solid TME imposes perpetual acetyl-coenzyme A (CoA) carboxylase (ACC) activity, invoking lipid biogenesis and storage in TILs that opposes FAO. Using metabolic, lipidomic, and confocal imaging strategies, we find that restricting ACC rewires Tcell metabolism, enabling energy maintenance in TME stress. Limiting ACC activity potentiates a gene and phenotypic program indicative of Tcell longevity, engendering Tcells with increased survival and polyfunctionality, which sustains cancer control.

Utilization of plant-derived sugars and lipids are coupled during colonization of rhizoplane and rhizosphere by the fungus Metarhizium robertsii

Plant-derived sugars and lipids are key nutritional sources for plant associated fungi. However, the relationship between utilization of host-derived sugars and lipids during development of the symbiotic association remains unknown. Here we show that the fungus Metarhizium robertsii also needs plant-derived lipids to develop symbiotic relationship with plants. The fatty acid binding proteins FABP1 and FABP2 are important for utilization of plant-derived lipids as the deletion of Fabp1 and Fabp2 significantly reduced the ability of M. robertsii to colonize rhizoplane and rhizosphere of maize and Arabidopsis thaliana. Deleting Fabp1 and Fabp2 increased sugar utilization by upregulating six sugar transporters, and this explains why deleting the monosaccharide transporter gene Mst1, which plays an important role in utilization of plant-derived sugars, had no impact on the ability of the double-gene deletion mutant ΔFabp1::ΔFabp2 to colonize plant roots. FABP1 and FABP2 were also found in other plant-associated Metarhizium species, and they were highly expressed in the medium using the tomato root exudate as the sole carbon and nitrogen source, suggesting that they could be also important for these species to develop symbiotic relationship with plants. In conclusion, we discovered that utilization of plant-derived sugars and lipids are coupled during colonization of rhizoplane and rhizosphere by M. robertsii.

Pathogenesis of MASLD and MASH - role of insulin resistance and lipotoxicity.

Insulin resistance and lipotoxicity are extremely interconnected but fundamental in setting the stage for the development of MASLD/MASH. A comprehensive literature search was performed and key themes were synthesised to provide insight into the underlying molecular mechanisms of insulin resistance and lipotoxicity in the liver, muscle, pancreas and adipose tissue and how organ cross-talk is fundamental to driving disease pathogenesis. Classical thinking postulates that excess FFA load exceeds the storage capacity of adipose tissue, which is predicated upon both genetic and environmental factors. This results in insulin resistance and compensatory hyperinsulinaemia by pancreatic beta cells to overcome target organ insulin resistance. As adipocyte dysfunction worsens, not only are excess FFA delivered to other organs, including skeletal muscle, pancreas and liver but a pro-inflammatory milieu is established with increases in IL-6, TNF-α and changes in adipokine levels (increased leptin and decreased adiponectin). With increased intramuscular lipid accumulation, lipotoxic species decrease insulin signalling, reduce glucose uptake by downregulation of GLUT4 and decrease glycogen synthesis. With this additional reduced capacity, hyperglycaemia is further exacerbated and increased FFA are delivered to the liver. The liver has the largest capacity to oxidise fat and to adapt to these stressors and, therefore, has become the last line of defence for excess lipid storage and utilisation, the capacity of which may be impacted by genetic and environmental factors. However, when the liver can no longer keep up with increasing FFA delivery and DNL, lipotoxic species accumulate with ensuing mitochondrial dysfunction, increased ER stress, oxidant stress and inflammasome activation, all of which drive hepatocyte injury and apoptosis. The resulting wound healing response, marked by stellate cell activation, drives collagen accumulation, progressive fibrosis, and, ultimately, end organ failure and death. This vicious cycle and complex interplay between insulin resistance, hyperinsulinaemia, lipotoxicity and multi-directional cross-talk among different target organs are critical drivers of MASLD/MASH. Targeting tissue-specific insulin resistance and hyperinsulinaemia while decreasing FFA load (lipotoxicity) through dietary and lifestyle changes remain the best upstream interventions.

Impact of live feed substitution with formulated diets on the development, digestive capacity, biochemical composition, and rearing water quality of European lobster (Homarus gammarus, L.) larvae

The lobster aquaculture industry, constrained by limited access to viable and cost-effective feed sources, prompted an investigation into the potential of replacing live feeds with formulated diets for Homarus gammarus larval culture, exploring alternative protein ingredients. Four diets were tested on newly-hatched H. gammarus: live Artemia nauplii (ART), an extruded control diet with krill meal and fishmeal as main protein sources (CTRL), and diets with 15% of dietary protein replaced by shrimp waste meal (SWM) or black soldier fly meal (BSF).Results revealed significant diet-induced effects on digestive enzymes in H. gammarus larvae. ART-fed lobsters exhibited increased trypsin and amylase activities but lower lipase activity than those fed extruded diets, suggesting more efficient utilization of protein and carbohydrate in ART-fed larvae, while formulated diets compensated with efficient lipid utilization. BSF and SWM diets increased exo- and endochitinase activities, likely due to chitin presence. Additionally, CTRL and BSF-fed larvae exhibited elevated body glycogen content. ART-fed larvae displayed a significantly higher ratio of low DNA to high DNA bacteria in their rearing water, indicative of lower nutrient loading. Despite these diet-induced impacts, there was no dietary effect on growth and survival during H. gammarus larval development.The study suggests successful formulated feed utilization by homarid larvae, even with alternative ingredients like BSF and SWM, offering a promising solution to reduce reliance on live feeds in lobster aquaculture. Further research is crucial to assess the long-term implications of the observed physiological responses to the different diets in later life stages.

Mitochondrial fusion and altered beta-oxidation drive muscle wasting in a Drosophila cachexia model

Cancer cachexia is a tumour-induced wasting syndrome, characterised by extreme loss of skeletal muscle. Defective mitochondria can contribute to muscle wasting; however, the underlying mechanisms remain unclear. Using a Drosophila larval model of cancer cachexia, we observed enlarged and dysfunctional muscle mitochondria. Morphological changes were accompanied by upregulation of beta-oxidation proteins and depletion of muscle glycogen and lipid stores. Muscle lipid stores were also decreased in Colon-26 adenocarcinoma mouse muscle samples, and expression of the beta-oxidation gene CPT1A was negatively associated with muscle quality in cachectic patients. Mechanistically, mitochondrial defects result from reduced muscle insulin signalling, downstream of tumour-secreted insulin growth factor binding protein (IGFBP) homologue ImpL2. Strikingly, muscle-specific inhibition of Forkhead box O (FOXO), mitochondrial fusion, or beta-oxidation in tumour-bearing animals preserved muscle integrity. Finally, dietary supplementation with nicotinamide or lipids, improved muscle health in tumour-bearing animals. Overall, our work demonstrates that muscle FOXO, mitochondria dynamics/beta-oxidation and lipid utilisation are key regulators of muscle wasting in cancer cachexia.

Dietary calcifediol reduces mesenteric adiposity to the benefit of carcass growth independently of circulating vitamin D hormone in juvenile Atlantic salmon

Dietary vitamin D is required by cultivated fish for optimal growth, feed utilisation and bone health. The in vivo conversion of cholecalciferol (D3) to active vitamin D hormone (1,25-OH-D3) may be impaired in farm-raised fish fed modern feed formulations. Requiring fewer metabolic steps for its conversion, the use of dietary calcifediol (25-OH-D3) may mitigate against deficits in vitamin D metabolism.A 90-day experimental feeding trial was undertaken in juvenile (23 g) Atlantic salmon (Salmo salar) to test the efficacy of 25-OH-D3 added to a standard diet containing recommended levels of D3 (5640 IU). Test diets contained 100, 200 or 400 μg/kg added 25-OH-D3 against a no additive control diet. Effects on zootechnical performance, indices of adiposity, bone mineralisation, and circulating vitamin D metabolites were tested.Dietary 25-OH-D3 significantly reduced feed conversion ratio (FCR) and resulted in numerical increases in growth performance. 25-OH-D3 also significantly reduced mesenteric adiposity with concurrent increases in carcass weight. Levels of calcium, phosphorous, magnesium, zinc and manganese were not modulated in the vertebrae of fish receiving 25-OH-D3. Circulating 25-OH-D3 remained below detection limits and 1,25-OH-D3 was not significantly modulated by the dietary manipulations.The significant reduction of mesenteric adiposity to the benefit of carcass weight shows that dietary 25-OH-D3 improves the utilisation of lipids. The vitamin D requirement for bone mineralisation was met by the basal diet. Segmented regression estimates a dietary 25-OH-D3 supplementation of 143 to 180 μg/kg for improved feed conversion and reduced mesenteric adiposity. The lack of correlation between circulating vitamin D hormone and mesenteric adiposity indicates that lipid metabolism is regulated by localised tissue activation of vitamin D, rather than the endocrine functions of circulating 1,25-OH-D3.

Sphingolipid metabolism controls mammalian heart regeneration

Utilization of lipids as energy substrates after birth causes cardiomyocyte (CM) cell-cycle arrest and loss of regenerative capacity in mammalian hearts. Beyond energy provision, proper management of lipid composition is crucial for cellular and organismal health, but its role in heart regeneration remains unclear. Here, we demonstrate widespread sphingolipid metabolism remodeling in neonatal hearts after injury and find that SphK1 and SphK2, isoenzymes producing the same sphingolipid metabolite sphingosine-1-phosphate (S1P), differently regulate cardiac regeneration. SphK2 is downregulated during heart development and determines CM proliferation via nuclear S1P-dependent modulation of histone acetylation. Reactivation of SphK2 induces adult CM cell-cycle re-entry and cytokinesis, thereby enhancing regeneration. Conversely, SphK1 is upregulated during development and promotes fibrosis through an S1P autocrine mechanism in cardiac fibroblasts. By fine-tuning the activity of each SphK isoform, we develop a therapy that simultaneously promotes myocardial repair and restricts fibrotic scarring to regenerate the infarcted adult hearts.

Mfn2/Hsc70 Complex Mediates the Formation of Mitochondria-Lipid Droplets Membrane Contact and Regulates Myocardial Lipid Metabolism.

The heart primarily derives its energy through lipid oxidation. In cardiomyocytes, lipids are stored in lipid droplets (LDs) and are utilized in mitochondria, although the structural and functional connections between these two organelles remain largely unknown. In this study, visible evidence have presented indicating that a complex is formed at the mitochondria-LD membrane contact (MLC) site, involving mitochondrion-localized Mfn2 and LD-localized Hsc70. This complex serves to tether mitochondria to LDs, facilitating the transfer of fatty acids (FAs) from LDs to mitochondria for β-oxidation. Reduction of Mfn2 induced by lipid overload inhibits MLC, hinders FA transfer, and results in lipid accumulation. Restoring Mfn2 reinstates MLC, alleviating myocardial lipotoxicity under lipid overload conditions both in-vivo and in-vitro. Additionally, prolonged lipid overload induces Mfn2 degradation through the ubiquitin-proteasome pathway, following Mfn2 acetylation at the K243 site. This leads to the transition from adaptive lipid utilization to maladaptive lipotoxicity. The experimental findings are supported by clinical data from patients with obesity and age-matched non-obese individuals. These translational results make a significant contribution to the molecular understanding of MLC in the heart, and offer new insights into its role in myocardial lipotoxicity.

Itaconate stabilizes CPT1a to enhance lipid utilization during inflammation

One primary metabolic manifestation of inflammation is the diversion of cis-aconitate within the tricarboxylic acid (TCA) cycle to synthesize the immunometabolite itaconate. Itaconate is well established to possess immunomodulatory and metabolic effects within myeloid cells and lymphocytes, however, its effects in other organ systems during sepsis remain less clear. Utilizing Acod1 knockout mice that are deficient in synthesizing itaconate, we aimed to understand the metabolic role of itaconate in the liver and systemically during sepsis. We find itaconate aids in lipid metabolism during sepsis. Specifically, Acod1 KO mice develop a heightened level of hepatic steatosis when induced with polymicrobial sepsis. Proteomics analysis reveals enhanced expression of enzymes involved in fatty acid oxidation in following 4-octyl itaconate (4-OI) treatment in vitro. Downstream analysis reveals itaconate stabilizes the expression of the mitochondrial fatty acid uptake enzyme CPT1a, mediated by its hypoubiquitination. Chemoproteomic analysis revealed itaconate interacts with proteins involved in protein ubiquitination as a potential mechanism underlying its stabilizing effect on CPT1a. From a systemic perspective, we find itaconate deficiency triggers a hypothermic response following endotoxin stimulation, potentially mediated by brown adipose tissue (BAT) dysfunction. Finally, by use of metabolic cage studies, we demonstrate Acod1 KO mice rely more heavily on carbohydrates versus fatty acid sources for systemic fuel utilization in response to endotoxin treatment. Our data reveal a novel metabolic role of itaconate in modulating fatty acid oxidation during polymicrobial sepsis.

Itaconate stabilizes CPT1a to enhance lipid utilization during inflammation.

One primary metabolic manifestation of inflammation is the diversion of cis-aconitate within the tricarboxylic acid (TCA) cycle to synthesize the immunometabolite itaconate. Itaconate is well established to possess immunomodulatory and metabolic effects within myeloid cells and lymphocytes, however, its effects in other organ systems during sepsis remain less clear. Utilizing Acod1 knockout mice that are deficient in synthesizing itaconate, we aimed to understand the metabolic role of itaconate in the liver and systemically during sepsis. We find itaconate aids in lipid metabolism during sepsis. Specifically, Acod1 KO mice develop a heightened level of hepatic steatosis when induced with polymicrobial sepsis. Proteomics analysis reveals enhanced expression of enzymes involved in fatty acid oxidation in following 4-octyl itaconate (4-OI) treatment in vitro. Downstream analysis reveals itaconate stabilizes the expression of the mitochondrial fatty acid uptake enzyme CPT1a, mediated by its hypoubiquitination. Chemoproteomic analysis revealed itaconate interacts with proteins involved in protein ubiquitination as a potential mechanism underlying its stabilizing effect on CPT1a. From a systemic perspective, we find itaconate deficiency triggers a hypothermic response following endotoxin stimulation, potentially mediated by brown adipose tissue (BAT) dysfunction. Finally, by use of metabolic cage studies, we demonstrate Acod1 KO mice rely more heavily on carbohydrates versus fatty acid sources for systemic fuel utilization in response to endotoxin treatment. Our data reveal a novel metabolic role of itaconate in modulating fatty acid oxidation during polymicrobial sepsis.

A proton birdcage coil integrated with interchangeable single loops for multi-nuclear MRI/MRS.

Energy metabolism is fundamental for life. It encompasses the utilization of carbohydrates, lipids, and proteins for internal processes, while aberrant energy metabolism is implicated in many diseases. In the present study, using three-dimensional (3D) printing from polycarbonate via fused deposition modeling, we propose a multi-nuclear radiofrequency (RF) coil design with integrated 1H birdcage and interchangeable X-nuclei (2H, 13C, 23Na, and 31P) single-loop coils for magnetic resonance imaging (MRI)/magnetic resonance spectroscopy (MRS). The single-loop coil for each nucleus attaches to an arc bracket that slides unrestrictedly along the birdcage coil inner surface, enabling convenient switching among various nuclei and animal handling. Compared to a commercial 1H birdcage coil, the proposed 1H birdcage coil exhibited superior signal-excitation homogeneity and imaging signal-to-noise ratio (SNR). For X-nuclei study, prominent peaks in spectroscopy for phantom solutions showed excellent SNR, and the static and dynamic peaks of in vivo spectroscopy validated the efficacy of the coil design in structural imaging and energy metabolism detection simultaneously.

Ceramides are fuel gauges on the drive to cardiometabolic disease.

Ceramides are signals of fatty acid excess that accumulate when a cell's energetic needs have been met and its nutrient storage has reached capacity. As these sphingolipids accrue, they alter the metabolism and survival of cells throughout the body including in the heart, liver, blood vessels, skeletal muscle, brain, and kidney. These ceramide actions elicit the tissue dysfunction that underlies cardiometabolic diseases such as diabetes, coronary artery disease, metabolic-associated steatohepatitis, and heart failure. Here, we review the biosynthesis and degradation pathways that maintain ceramide levels in normal physiology and discuss how the loss of ceramide homeostasis drives cardiometabolic pathologies. We highlight signaling nodes that sense small changes in ceramides and in turn reprogram cellular metabolism and stimulate apoptosis. Finally, we evaluate the emerging therapeutic utility of these unique lipids as biomarkers that forecast disease risk and as targets of ceramide-lowering interventions that ameliorate disease.

Roles of Myokines and Muscle-Derived Extracellular Vesicles in Musculoskeletal Deterioration under Disuse Conditions.

Prolonged inactivity and disuse conditions, such as those experienced during spaceflight and prolonged bedrest, are frequently accompanied by detrimental effects on the motor system, including skeletal muscle atrophy and bone loss, which greatly increase the risk of osteoporosis and fractures. Moreover, the decrease in glucose and lipid utilization in skeletal muscles, a consequence of muscle atrophy, also contributes to the development of metabolic syndrome. Clarifying the mechanisms involved in disuse-induced musculoskeletal deterioration is important, providing therapeutic targets and a scientific foundation for the treatment of musculoskeletal disorders under disuse conditions. Skeletal muscle, as a powerful endocrine organ, participates in the regulation of physiological and biochemical functions of local or distal tissues and organs, including itself, in endocrine, autocrine, or paracrine manners. As a motor organ adjacent to muscle, bone tissue exhibits a relative lag in degenerative changes compared to skeletal muscle under disuse conditions. Based on this phenomenon, roles and mechanisms involved in the communication between skeletal muscle and bone, especially from muscle to bone, under disuse conditions have attracted widespread attention. In this review, we summarize the roles and regulatory mechanisms of muscle-derived myokines and extracellular vesicles (EVs) in the occurrence of muscle atrophy and bone loss under disuse conditions, as well as discuss future perspectives based on existing research.

Genetic factors affecting storage and utilization of lipids during dormancy in Mycobacterium tuberculosis.

Mycobacterium tuberculosis (Mtb) can adopt a non-growing dormant state during infection that may be critical to both active and latent tuberculosis. During dormancy, Mtb is widely tolerant toward antibiotics, a significant obstacle in current anti-tubercular drug regimens, and retains the ability to persist in its environment. We aimed to identify novel mechanisms that permit Mtb to survive dormancy in an in vitro carbon starvation model using transposon insertion sequencing and gene expression analysis. We identified a previously uncharacterized component of the lipid transport machinery, omamC, which was upregulated and required for survival during carbon starvation. We show that OmamC plays a role both in increasing fatty acid stores during growth in rich media and enhancing fatty acid utilization during starvation. Besides its involvement in lipid metabolism, OmamC levels affected the expression of the anti-anti-sigma factor rv0516c and other genes to improve Mtb survival during carbon starvation and increase its tolerance toward rifampicin, a first-line drug effective against non-growing Mtb. Importantly, we show that Mtb can be eradicated during carbon starvation, in an OmamC-dependent manner, by inhibiting lipid metabolism with the lipase inhibitor tetrahydrolipstatin. This work casts new light into the survival processes of non-replicating, drug-tolerant Mtb by identifying new proteins involved in lipid metabolism required for the survival of dormant bacteria and exposing a potential vulnerability that could be exploited for antibiotic discovery.IMPORTANCETuberculosis is a global threat, with ~10 million yearly active cases. Many more people, however, live with "latent" infection, where Mycobacterium tuberculosis survives in a non-replicative form. When latent bacteria activate and regrow, they elicit immune responses and result in significant host damage. Replicating and non-growing bacilli can co-exist; however, non-growing bacteria are considerably less sensitive to antibiotics, thus complicating treatment by necessitating long treatment durations. Here, we sought to identify genes important for bacterial survival in this non-growing state using a carbon starvation model. We found that a previously uncharacterized gene, omamC, is involved in storing and utilizing fatty acids as bacteria transition between these two states. Importantly, inhibiting lipid metabolism using a lipase inhibitor eradicates non-growing bacteria. Thus, targeting lipid metabolism may be a viable strategy for treating the non-growing population in strategies to shorten treatment durations of tuberculosis.

Dietary oleic acid intake increases the proportion of type 1 and 2X muscle fibers in mice

Skeletal muscle is one of the largest metabolic tissues in mammals and is composed of four different types of muscle fibers (types 1, 2A, 2X, and 2B); however, type 2B is absent in humans. Given that slow-twitch fibers are superior to fast-twitch fibers in terms of oxidative metabolism and are rich in mitochondria, shift of muscle fiber types in direction towards slower fiber types improves metabolic disorders and endurance capacity. We previously had reported that oleic acid supplementation increases type 1 fiber formation in C2C12 myotubes; however, its function still remains unclear. This study aimed to determine the effect of oleic acid on the muscle fiber types and endurance capacity. An in vivo mouse model was used, and mice were fed a 10% oleic acid diet for 4 weeks. Two different skeletal muscles, slow soleus muscle with the predominance of slow-twitch fibers and fast extensor digitorum longus (EDL) muscle with the predominance of fast-twitch fibers, were used. We found that dietary oleic acid intake improved running endurance and altered fiber type composition of muscles, the proportion of type 1 and 2X fibers increased in the soleus muscle and type 2X increased in the EDL muscle. The fiber type shift in the EDL muscle was accompanied by an increased muscle TAG content. In addition, blood triacylglycerol (TAG) and non-esterified fatty acid levels decreased during exercise. These changes suggested that lipid utilization as an energy substrate was enhanced by oleic acid. Increased proliferator-activated receptor γ coactivator-1β protein levels were observed in the EDL muscle, which potentially enhanced the fiber type transitions towards type 2X and muscle TAG content. In conclusion, dietary oleic acid intake improved running endurance with the changes of muscle fiber type shares in mice. This study elucidated a novel functionality of oleic acid in skeletal muscle fiber types. Further studies are required to elucidate the underlying mechanisms. Our findings have the potential to contribute to the field of health and sports science through nutritional approaches, such as the development of supplements aimed at improving muscle function.

Probing Leishmania donovani Metabolism in 3D Culture: New Perspectives on Drug Targets

Objective: The goal of the research was to investigate Leishmania donovani (L. donovani) metabolism in a 3D culture environment and find possible targets for metabolic drugs. Methods: Different metabolic experiments were performed on two strains of L. donovani (LD1S and LD2B) that were cultivated in 3D spheroids. In order to comprehend the metabolic processes of L. donovani, glucose intake, lactate generation, and lipid utilization were evaluated. Hexokinase (HK) and fatty acid synthase (FAS), two important enzymes implicated in these metabolic pathways, were recognized as potential therapeutic targets. HK and FAS were knocked down in the parasite using CRISPR-Cas9 technology to validate these targets, and the effects on growth and viability were statistically evaluated. Results: In the 3D culture, L. donovani showed a strong growth pattern and significantly increased in diameter over the course of 14 days (p<0.001). When compared to earlier 2D investigations, metabolic tests revealed that 3D cells consumed much more glucose and used lipids (p<0.001). In comparison to wild-type strains, HK and FAS knockdown strains drastically decreased their use of lipids and glucose, respectively (p<0.001). In comparison to the wild-type strains, both knockdown variants demonstrated significantly less growth and vitality (p<0.001). Conclusion: Our results demonstrate the value of 3D culture for investigating the metabolism of L. donovani, and they also identify and verify HK and FAS as prospective therapeutic targets. These discoveries could open the door for the creation of novel treatment approaches to treat L. donovani infection. The parasite's considerable growth and survival decrease when these metabolic pathways are interrupted is supported by statistical evidence, opening the door for future therapeutic development.

Artemether regulates liver glycogen and lipid utilization through mitochondrial pyruvate oxidation in db/db mice

Artemether regulates liver glycogen and lipid utilization through mitochondrial pyruvate oxidation in db/db mice