Accumulation Of Droplets Research Articles

LEP inhibits intramuscular adipogenesis through the AMPK signaling pathway invitro.

Leptin can indirectly regulate fatty-acid metabolism and synthesis in muscle invivo and directly in incubated muscle exvivo. In addition, non-synonymous mutations in the bovine leptin gene (LEP) are associated with carcass intramuscular fat (IMF) content. However, the effects of LEP on lipid synthesis of adipocytes have not been clearly studied at the cellular level. Therefore, this study focused on bovine primary intramuscular preadipocytes to investigate the effects of LEP on the proliferation and differentiation of intramuscular preadipocytes, as well as its regulatory mechanism in lipid synthesis. The results showed that both the LEP and leptin receptor gene (LEPR) were highly expressed in IMF tissues, and their mRNA expression levels were positively correlated at different developmental stages of intramuscular preadipocytes. The overexpression of LEP inhibited the proliferation and differentiation of intramuscular preadipocytes, while interference with LEP had the opposite effect. Additionally, LEP significantly promoted the phosphorylation level of AMPKα by promoting the protein expression of CAMKK2. Meanwhile, rescue experiments showed that the increasing effect of AMPK inhibitors on the number of intramuscular preadipocytes was significantly weakened by the overexpression of LEP. Furthermore, the overexpression of LEP could weaken the promoting effect of AMPK inhibitor on triglyceride content and droplet accumulation, and prevent the upregulation of adipogenic protein expression (SREBF1, FABP4, FASN, and ACCα) caused by AMPK inhibitor. Taken together, LEP acted on the AMPK signaling pathway by regulating the protein expression of CAMKK2, thereby downregulating the expression of proliferation-related and adipogenic-related genes and proteins, ultimately reducing intramuscular adipogenesis.

Investigating the Effects of Lubricant Infusion Methods on Polymer SLIPS.

Slippery lubricant infused porous surfaces (SLIPSs) are promising bioinspired surfaces with self-healing and droplet wetting properties, among many others, that are desirable due to their range of applications. Recently, there have been many developments in the SLIPS field regarding the creation of textured surfaces and lubricant selection. However, there is a lack of knowledge regarding the method of lubricant infusion. In this study, we aim to fill this void by investigating different infusion methods that impose external forces on the lubricant. We developed our SLIPS by hot embossing nanostructures onto polypropylene by using molds that were laser micromachined. These textured surfaces were then infused with silicone oil using three different infusion methods: ultrasonication, vacuum, and hydrostatic pressure. We analyzed the wettability and slipperiness of the SLIPS by evaluating the critical tilt angle and comparing the sliding velocities of water droplets on each sample at a tilt angle of 20°. Additionally, the durability of the SLIPS was tested by dropping 50 successive water drops onto the samples and evaluating the droplet-surface interactions throughout. The sonicated infusion method yielded SLIPS that performed the best with a contact angle hysteresis of 13°, a critical tilt angle of 18.3°, a sliding velocity of 1.66 mm/s, and the least accumulation of droplets over time with use. These values are greatly improved when compared to the control sample where lubricant was simply dripped on, which resulted in a contact angle hysteresis of 20°, a critical tilt angle of 26.3°, and a sliding velocity of 0.23 mm/s. The sonicated and drip infusion methods were also compared with different materials (stainless steel) and different textures (microstructures). It was found that the improvement in slipperiness using the sonicated infusion method is prominent for nanoscale textures on both stainless steel and polypropylene. In this study, we discuss the challenges with oil depletion in SLIPS (cloaking and wetting ridges) and with the selection of contact angle measurement methods. While further investigation as to why certain applied forces during infusion yield better SLIPS is warranted, these forces greatly affect the outcome. This work suggests that researchers should consider using sonication or other methods of lubricant infusion that apply external forces as infusion techniques to yield better SLIPS on the nanoscale.

Adjusting droplet adhesion of superhydrophobic coating via surface embedding of microparticles with mixed shapes

Designing intelligent superhydrophobic surfaces with versatile surface properties has garnered scholarly attention in field of heterogeneous catalysis, lab-on-chip, water harvesting, etc. However, the precise modulation of fluid adhesion on superhydrophobic surfaces, particularly when being applied as coatings, remains a challenge. Here we employ a “glue + powder” method for the fabrication of superhydrophobic surfaces with tunable droplet adhesion by embedding microparticles with mixed shapes. By adjusting the mixing ratio of rod-like and spherical particles, the sliding angle of superhydrophobic coating can be facilely controlled as well as the underwater bubble holding ability. During the dewetting process, the difference in the contact line lengths between micro-rod and micro-sphere contributes to the varying droplet adhesive force from 43.7 μN to 109.4 μN, which corresponds to the rolling-off angle change of a 10 μL droplet from 9.6° to 45.7°. Furthermore, by mixing fumed silica with branched structure, the lowest range of the rolling-off angle adjustment can be extended down to 0.5°, showing a precise and universal strategy for tuning the surface adhesion. On the basis of the meticulous manipulation of surface adhesion, controlling processes of single droplet and droplets accumulation can be fulfilled, with implications for enhancing the functionality of current superhydrophobic materials. Taking the advantage of the convenient and reliable fabrication method, a series of demonstration including manual droplet holders, multi-tiered droplet capturing interface, droplet sieving platforms, and gravity-driven droplet reactors has been realized. We envision that the precise control of droplet adhesion on superhydrophobic materials should further provides substantial opportunities for innovating fluid manipulation systems possessing heightened operability and multifunctionality.

Effect and application of spray drying and freeze drying on characterization of walnut oil microcapsules

In the current study, a walnut oil (WO) emulsion was prepared with WO as the core material and sodium caseinate and maltodextrin as the wall materials. The effects of spray drying (SD) and freeze drying (FD) on the structure and quality of WO microcapsules were compared, and the role of WO microcapsules as a substitute for commercial creamer in coffee was investigated. The results demonstrated that the WO microcapsules prepared by the SD method had dense microstructure, spherical particles, no oil drop aggregation, small particle size, good fluidity, an embedding rate of up to 93.60%, an unsaturated fatty acid content of 87.00%, and no significant difference in thermal stability or stability of the restored emulsion compared with FD. The freeze-dried WO microcapsules have irregular structure, porous surface structure, high oil content on the surface, 76.84% embedding rate, some accumulation of droplets, large grain size, and poor fluidity. The electronic nose and electronic tongue results reveal that the microcapsules prepared by the two methods have similar flavor when applied to coffee. The spray-dried WO microcapsules are prepared with a flavor closer to that of commercial creams. In summary, microcapsules prepared by SD are superior to FD and are promising for industrial production; further, the nondairy creamer based on WO has great potential for coffee application.

Ultrafast Droplets Trajectories Modulation of Electrohydrodynamic Jet Printing for Pattern Refinement Using Electrostatic Deflection

AbstractElectrohydrodynamic jet (E‐Jet) printing is a highly promising non‐contact additive manufacturing technology. However, the fast jets generated by E‐Jet may cause droplet accumulation on small curvature features due to the slow motion control achieved by the mechanical stage, leading to a degradation of printed patterns in localized areas. Here, a method is proposed for ultrafast modulation of jet trajectories to achieve finely patterned E‐Jet printing by introducing jet‐deflecting electrodes around the jet, which can continuously transform the jet trajectory with lateral acceleration up to 104 ms−2. This extraordinary acceleration is two orders of magnitude higher than that can be achieved by using mechanical stage motion. Through a combination of numerical calculations and experiments, the effects of electric field parameters on the feature size and kinematic properties of jet droplets are investigated. In addition, deposition errors of <5 µm are fabricated by selecting appropriate working parameters and using UV‐curable adhesives as printing ink. Subsequently, complex micropatterns with feature curvatures <4 µm are successfully printed, demonstrating the consistency and reliability of this electrostatic deflection technique in achieving a uniform droplet deposition pitch. This innovative approach holds significant promise in the field of high‐quality microscale additive manufacturing.

Emission spectroscopy at high frame rates during ablation tests of meteorite samples in plasma wind tunnel

Optical emission spectra between 522–580 nm of ablating meteorites have been recorded at frame rates as high as 1 kHz for the first time during ground testing with simultaneous spatial and temporal resolution. A novel high frame rate emission spectroscopy arrangement has been developed and employed to diagnose the ablating meteorites in several experimental campaigns. In addition to the identification of species from emission lines detected, the resulting high-speed spectral data were used to study the temporal and spatial evolution of melting droplets and the associated spectral signatures. The time history of radiance from the atomic species emission was used to interpret the fragmentation behavior of various meteorites. Chelyabinsk meteorite exhibit almost constant radiance over time indicating steady droplet detachment whereas Ragland meteorite shows infrequent radiance peaks corresponding to random fragmentation/droplet detachment of varying sizes. A gradual rise in radiance history from iron meteorite Mount Joy shows that it takes finite time for melting and accumulation of droplets.

Proposal of a new slit-lamp shield for ophthalmic examination and assessment of its effectiveness using computational simulations.

This study aimed to use computational models for simulating the movement of respiratory droplets when assessing the efficacy of standard slit-lamp shield versus a new shield designed for increased clinician comfort as well as adequate protection. Simulations were performed using the commercial software Star-CCM+. Respiratory droplets were assumed to be 100% water in volume fraction with particle diameter distribution represented by a geometric mean of 74.4 (±1.5 standard deviation) μm over a 4-min duration. The total mass of respiratory droplets expelled from patients' mouths and droplet accumulation on the manikin were measured under the following three conditions: with no slit-lamp shield, using the standard slit-lamp shield, and using our new proposed shield. The total accumulated water droplet mass (kilogram) and percentage of expelled mass accumulated on the shield under the three aforementioned conditions were as follows: 5.84e-10 kg (28% of the total weight of particle emitted that settled on the manikin), 9.14e-13 kg (0.045%), and 3.19e-13 (0.015%), respectively. The standard shield could shield off 99.83% of the particles that would otherwise be deposited on the manikin, which is comparable to 99.95% for the proposed design. Conclusion: Slit-lamp shields are effective infection control tools against respiratory droplets. The proposed shield showed comparable effectiveness compared with conventional slit-lamp shields, but with potentially enhanced ergonomics for ophthalmologists during slit-lamp examinations.

Glioblastoma disrupts the ependymal wall and extracellular matrix structures of the subventricular zone

BackgroundGlioblastoma (GBM) is the most aggressive and common type of primary brain tumor in adults. Tumor location plays a role in patient prognosis, with tumors proximal to the lateral ventricles (LVs) presenting with worse overall survival, increased expression of stem cell genes, and increased incidence of distal tumor recurrence. This may be due in part to interaction of GBM with factors of the subventricular zone (SVZ), including those contained within the cerebrospinal fluid (CSF). However, direct interaction of GBM tumors with CSF has not been proved and would be hindered in the presence of an intact ependymal cell layer.MethodsHere, we investigate the ependymal cell barrier and its derived extracellular matrix (ECM) fractones in the vicinity of a GBM tumor. Patient-derived GBM cells were orthotopically implanted into immunosuppressed athymic mice in locations distal and proximal to the LV. A PBS vehicle injection in the proximal location was included as a control. At four weeks post-xenograft, brain tissue was examined for alterations in ependymal cell health via immunohistochemistry, scanning electron microscopy, and transmission electron microscopy.ResultsWe identified local invading GBM cells within the LV wall and increased influx of CSF into the LV-proximal GBM tumor bulk compared to controls. In addition to the physical disruption of the ependymal cell barrier, we also identified increased signs of compromised ependymal cell health in LV-proximal tumor-bearing mice. These signs include increased accumulation of lipid droplets, decreased cilia length and number, and decreased expression of cell channel proteins. We additionally identified elevated numbers of small fractones in the SVZ within this group, suggesting increased indirect CSF-contained molecule signaling to tumor cells.ConclusionsOur data is the first to show that LV-proximal GBMs physically disrupt the ependymal cell barrier in animal models, resulting in disruptions in ependymal cell biology and increased CSF interaction with the tumor bulk. These findings point to ependymal cell health and CSF-contained molecules as potential axes for therapeutic targeting in the treatment of GBM.

Diacylglycerol acyltransferase 2 promotes the adipogenesis of intramuscular preadipocytes in goat

Diacylglycerol acyltransferase 2 (DGAT2) is the key enzyme that catalyzes the last step of triglyceride synthesis. However, its role in intramuscular fat (IMF) deposition in goat remains unclear. The purpose of this study was to explore the role of DGAT2 in regulating goat IMF deposition. In the present study, the expression of DGAT2 was highest in goat triceps brachii, and highest on the first day after oleic acid induction in goat intramuscular preadipocytes. The overexpression of DGAT2 promoted the accumulation of lipid droplets and triglyceride synthesis, accompanied by the expression upregulation of DGAT1, TIP47, ACC and ACOX1 significantly, and expression downregulation of AGPAT6, LPIN1, LPL, HSL, ATGL and ADRP significantly. In contrast, the silencing of DGAT2 decreased the accumulation of lipid droplets, inhibited the expression of DGAT1, GPAM, ADRP, AGPAT6, LPL, HSL, ATGL, ACC, FASN, ACOX1 significantly, and enhanced that of TIP47 significantly. Overall, these data underscore DGAT2 may play a potentially important role in lipid droplets formation and triglyceride accumulation, so as to maintain intramuscular fat deposition, beyond triglyceride synthesis in goat.

Prostaglandin 2α Promotes Autophagy and Mitochondrial Energy Production in Fish Hepatocytes.

Fatty liver, characterized by excessive lipid droplet (LD) accumulation in hepatocytes, is a common physiological condition in humans and aquaculture species. Lipid mobilization is an important strategy for modulating the number and size of cellular LDs. Cyclooxygenase (COX)-mediated arachidonic acid derivatives are known to improve lipid catabolism in fish; however, the specific derivatives remain unknown. In the present study, we showed that serum starvation induced LD degradation via autophagy, lipolysis, and mitochondrial energy production in zebrafish hepatocytes, accompanied by activation of the COX pathway. The cellular concentration of PGF2α, but not other prostaglandins, was significantly increased. Administration of a COX inhibitor or interference with PGF2α synthase abolished serum deprivation-induced LD suppression, LD–lysosome colocalization, and expression of autophagic genes. Additionally, exogenous PGF2α suppressed the accumulation of LDs, promoted the accumulation of lysosomes with LD and the autophagy marker protein LC3A/B, and augmented the expression of autophagic genes. Moreover, PGF2α enhanced mitochondrial accumulation and ATP production, and increased the transcript levels of β-oxidation- and mitochondrial respiratory chain-related genes. Collectively, these findings demonstrate that the COX pathway is implicated in lipid degradation induced by energy deprivation, and that PGF2α is a key molecule triggering autophagy, lipolysis, and mitochondrial development in zebrafish hepatocytes.

Lipid droplet degradation by autophagy connects mitochondria metabolism to Prox1-driven expression of lymphatic genes and lymphangiogenesis

Autophagy has vasculoprotective roles, but whether and how it regulates lymphatic endothelial cells (LEC) homeostasis and lymphangiogenesis is unknown. Here, we show that genetic deficiency of autophagy in LEC impairs responses to VEGF-C and injury-driven corneal lymphangiogenesis. Autophagy loss in LEC compromises the expression of main effectors of LEC identity, like VEGFR3, affects mitochondrial dynamics and causes an accumulation of lipid droplets (LDs) in vitro and in vivo. When lipophagy is impaired, mitochondrial ATP production, fatty acid oxidation, acetyl-CoA/CoA ratio and expression of lymphangiogenic PROX1 target genes are dwindled. Enforcing mitochondria fusion by silencing dynamin-related-protein 1 (DRP1) in autophagy-deficient LEC fails to restore LDs turnover and lymphatic gene expression, whereas supplementing the fatty acid precursor acetate rescues VEGFR3 levels and signaling, and lymphangiogenesis in LEC-Atg5−/− mice. Our findings reveal that lipophagy in LEC by supporting FAO, preserves a mitochondrial-PROX1 gene expression circuit that safeguards LEC responsiveness to lymphangiogenic mediators and lymphangiogenesis.

L-theanine prevents progression of nonalcoholic hepatic steatosis by regulating hepatocyte lipid metabolic pathways via the CaMKK\u03b2-AMPK signaling pathway

BackgroundL-theanine, a non-protein amino acid was found principally in the green tea, has been previously shown to exhibit potent anti-obesity property and hepatoprotective effect. Herein, we investigated the effects of L-theanine on alleviating nonalcoholic hepatic steatosis in vitro and in vivo, and explored the underlying molecular mechanism.MethodsIn vitro, HepG2 and AML12 cells were treated with 500 μM oleic acid (OA) or treated with OA accompanied by L-theanine. In vivo, C57BL/6J mice were fed with normal control diet (NCD), high‐fat diet (HFD), or HFD along with L-theanine for 16 weeks. The levels of triglycerides (TG), accumulation of lipid droplets and the expression of genes related to hepatocyte lipid metabolic pathways were detected in vitro and in vivo.ResultsOur data indicated that, in vivo, L-theanine significantly reduced body weight, hepatic steatosis, serum levels of alanine transaminase (ALT), aspartate transaminase (AST), TG and LDL cholesterol (LDL-C) in HFD-induced nonalcoholic fatty liver disease (NAFLD) mice. In vitro, L-theanine also significantly alleviated OA induced hepatocytes steatosis. Mechanic studies showed that L-theanine significantly inhibited the nucleus translocation of sterol regulatory element binding protein 1c (SREBP-1c) through AMPK-mTOR signaling pathway, thereby contributing to the reduction of fatty acid synthesis. We also identified that L-theanine enhanced fatty acid β-oxidation by increasing the expression of peroxisome proliferator–activated receptor α (PPARα) and carnitine palmitoyltransferase-1 A (CPT1A) through AMP-activated protein kinase (AMPK). Furthermore, our study indicated that L-theanine can active AMPK through its upstream kinase Calmodulin-dependent protein kinase kinase-β (CaMKKβ).ConclusionsTaken together, our findings suggested that L-theanine alleviates nonalcoholic hepatic steatosis by regulating hepatocyte lipid metabolic pathways via the CaMKKβ-AMPK signaling pathway.

In vitro ballooned hepatocytes can be produced by primary human hepatocytes and hepatic stellate cell sheets

Despite the increasing prevalence of Nonalcoholic steatohepatitis (NASH) worldwide, there is no effective treatment available for this disease. “Ballooned hepatocyte” is a characteristic finding in NASH and is correlated with disease prognosis, but their mechanisms of action are poorly understood; furthermore, neither animal nor in vitro models of NASH have been able to adequately represent ballooned hepatocytes. Herein, we engineered cell sheets to develop a new in vitro model of ballooned hepatocytes. Primary human hepatocytes (PHH) and Hepatic stellate cells (HSC) were co-cultured to produce cell sheets, which were cultured in glucose and lipid containing medium, following which histological and functional analyses were performed. Histological findings showed hepatocyte ballooning, accumulation of fat droplets, abnormal cytokeratin arrangement, and the presence of Mallory–Denk bodies and abnormal organelles. These findings are similar to those of ballooned hepatocytes in human NASH. Functional analysis showed elevated levels of TGFβ-1, SHH, and p62, but not TNF-α, IL-8. Exposure of PHH/HSC sheets to a glucolipotoxicity environment induces ballooned hepatocyte without inflammation. Moreover, fibrosis is an important mechanism underlying ballooned hepatocytes and could be the basis for the development of a new in vitro NASH model with ballooned hepatocytes.

Liver X receptor-agonist treatment rescues degeneration in a Drosophila model of hereditary spastic paraplegia

Hereditary spastic paraplegias (HSPs) are a group of inherited, progressive neurodegenerative conditions characterised by prominent lower-limb spasticity and weakness, caused by a length-dependent degeneration of the longest corticospinal upper motor neurons. While more than 80 spastic paraplegia genes (SPGs) have been identified, many cases arise from mutations in genes encoding proteins which generate and maintain tubular endoplasmic reticulum (ER) membrane organisation. The ER-shaping proteins are essential for the health and survival of long motor neurons, however the mechanisms by which mutations in these genes cause the axonopathy observed in HSP have not been elucidated. To further develop our understanding of the ER-shaping proteins, this study outlines the generation of novel in vivo and in vitro models, using CRISPR/Cas9-mediated gene editing to knockout the ER-shaping protein ADP-ribosylation factor-like 6 interacting protein 1 (ARL6IP1), mutations in which give rise to the HSP subtype SPG61. Loss of Arl6IP1 in Drosophila results in progressive locomotor deficits, emulating a key aspect of HSP in patients. ARL6IP1 interacts with ER-shaping proteins and is required for regulating the organisation of ER tubules, particularly within long motor neuron axons. Unexpectedly, we identified physical and functional interactions between ARL6IP1 and the phospholipid transporter oxysterol-binding protein-related protein 8 in both human and Drosophila model systems, pointing to a conserved role for ARL6IP1 in lipid homeostasis. Furthermore, loss of Arl6IP1 from Drosophila neurons results in a cell non-autonomous accumulation of lipid droplets in axonal glia. Importantly, treatment with lipid regulating liver X receptor-agonists blocked lipid droplet accumulation, restored axonal ER organisation, and improved locomotor function in Arl6IP1 knockout Drosophila. Our findings indicate that disrupted lipid homeostasis contributes to neurodegeneration in HSP, identifying a potential novel therapeutic avenue for the treatment of this disorder.

Analysis of flame propagation mechanisms during light-round in an annular spray flame combustor: the impact of wall heat transfer and two-phase flow

Ignition in annular multi-burner combustors is marked by a succession of four phases, ending with a characteristic flame expansion from burner to burner, often referred to as light-round. During this last phase, flame propagation is prone to substantial change depending on the boundary and operating conditions. With realistic aero-engine conditions in mind, wall heat transfers can be enhanced during ignition in cold wall conditions, which aid an understanding of the main governing mechanisms of flame propagation. From a modeling perspective, several works have outlined the need for detailed descriptions of the liquid phase, turbulent combustion and wall heat transfer, which are all included in the present work for the first time. Large-Eddy Simulations of light-round are performed in the annular MICCA-Spray combustor with cold walls, Lagrangian particle tracking, a dynamic closure for the sub-grid scale flame surface wrinkling as well as a custom tabulated wall model. The predicted light-round duration from the simulation is found to be in good agreement with experimental data. It is shown that the volumetric expansion of burnt gases induces a flow acceleration in azimuthal direction which constitutes the main driving mechanism of flame propagation. Droplet accumulations in the wake of swirling jets are generated ahead of the propagating flame fronts, which in turn cause a characteristic sawtooth propagation mode of the leading point. A cooling effect of the combustor walls on burnt gases is particularly pronounced downstream, diminishing the resulting flame propagation speed. The main governing mechanisms are investigated by means of a mathematical model for the absolute turbulent flame speed to quantify their relative impact on flame propagation. Finally, a priori estimations are provided for the flame propagation speed based on different models and boundary conditions, which are directly plugged into the model.

Effects of Electrode Negative Pulsing Ratio in Direct Energy Deposition via Variable-Polarity Cold Metal Transfer Process on the Deposition Behavior and Microstructural Characteristics

Interest in research on the application of variable-polarity cold metal transfer mode in wire-based direct energy deposition has been growing; particularly popular are investigations into the respective influences of polarity, amplitude of the arc current, and polarity variation sequence on the quality of the final product manufactured via additive manufacturing. The application of the electrode-negative phase is more capable of yielding relatively large droplets and increasing the weight of the deposited material. However, the proportions of the electrode positive phase are typically larger than those of the electrode-negative phase because it maintains arc stability and droplet transfer. This discrepancy has prevented the accurate evaluation of the effects of the polarity mode and polarity sequences on the deposition characteristics associated with variable-polarity cold metal transfer. In this study, variable-polarity cold metal transfer was performed using a tuned waveform, and the effects of the electrode-negative pulsing ratio and pulse repetition on the geometrical features and deposition rate were assessed. The weight tended to increase with decreasing welding speed and increasing electrode-negative pulsing ratio. The number of repetitions influenced molten pool behavior, and when sufficiently high, induced ripple formation via droplet accumulation below the electrode. In addition, the effects of the electrode-negative pulsing ratio and repetition on the microstructure formation were analyzed. It was revealed that the average grain size was related to the amount of supplied energy and polarity switching during grain formation.

Decrease of Intracellular Glutamine by STF-62247 Results in the Accumulation of Lipid Droplets in von Hippel-Lindau Deficient Cells.

Kidney cancer is one of the top ten cancer diagnosed worldwide and its incidence has increased the last 20 years. Clear Cell Renal Cell Carcinoma (ccRCC) are characterized by mutations that inactivate the von Hippel-Lindau (VHL) tumor suppressor gene and evidence indicated alterations in metabolic pathways, particularly in glutamine metabolism. We previously identified a small molecule, STF-62247, which target VHL-deficient renal tumors by affecting late-stages of autophagy and lysosomal signaling. In this study, we investigated ccRCC metabolism in VHL-deficient and proficient cells exposed to the small molecule. Metabolomics profiling using 1H NMR demonstrated that STF-62247 increases levels of glucose, pyruvate, glycerol 3-phosphate while glutamate, asparagine, and glutathione significantly decreased. Diminution of glutamate and glutamine was further investigated using mass spectrometry, western blot analyses, enzymatic activities, and viability assays. We found that expression of SLC1A5 increases in VHL-deficient cells treated with STF-62247, possibly to stimulate glutamine uptake intracellularly to counteract the diminution of this amino acid. However, exogenous addition of glutamine was not able to rescue cell viability induced by the small molecule. Instead, our results showed that VHL-deficient cells utilize glutamine to produce fatty acid in response to STF-62247. Surprisingly, this occurs through oxidative phosphorylation in STF-treated cells while control cells use reductive carboxylation to sustain lipogenesis. We also demonstrated that STF-62247 stimulated expression of stearoyl-CoA desaturase (SCD1) and peripilin2 (PLIN2) to generate accumulation of lipid droplets in VHL-deficient cells. Moreover, the carnitine palmitoyltransferase 1A (CPT1A), which control the entry of fatty acid into mitochondria for β-oxidation, also increased in response to STF-62247. CPT1A overexpression in ccRCC is known to limit tumor growth. Together, our results demonstrated that STF-62247 modulates cellular metabolism of glutamine, an amino acid involved in the autophagy-lysosome process, to support lipogenesis, which could be implicated in the signaling driving to cell death.

Histone acetyltransferase NAA40 modulates acetyl-CoA levels and lipid synthesis

BackgroundEpigenetic regulation relies on the activity of enzymes that use sentinel metabolites as cofactors to modify DNA or histone proteins. Thus, fluctuations in cellular metabolite levels have been reported to affect chromatin modifications. However, whether epigenetic modifiers also affect the levels of these metabolites and thereby impinge on downstream metabolic pathways remains largely unknown. Here, we tested this notion by investigating the function of N-alpha-acetyltransferase 40 (NAA40), the enzyme responsible for N-terminal acetylation of histones H2A and H4, which has been previously implicated with metabolic-associated conditions such as age-dependent hepatic steatosis and calorie-restriction-mediated longevity.ResultsUsing metabolomic and lipidomic approaches, we found that depletion of NAA40 in murine hepatocytes leads to significant increase in intracellular acetyl-CoA levels, which associates with enhanced lipid synthesis demonstrated by upregulation in de novo lipogenesis genes as well as increased levels of diglycerides and triglycerides. Consistently, the increase in these lipid species coincide with the accumulation of cytoplasmic lipid droplets and impaired insulin signalling indicated by decreased glucose uptake. However, the effect of NAA40 on lipid droplet formation is independent of insulin. In addition, the induction in lipid synthesis is replicated in vivo in the Drosophila melanogaster larval fat body. Finally, supporting our results, we find a strong association of NAA40 expression with insulin sensitivity in obese patients.ConclusionsOverall, our findings demonstrate that NAA40 affects the levels of cellular acetyl-CoA, thereby impacting lipid synthesis and insulin signalling. This study reveals a novel path through which histone-modifying enzymes influence cellular metabolism with potential implications in metabolic disorders.

Drosophila Lysophospholipase Gene swiss cheese Is Required for Survival and Reproduction.

Simple SummaryBiological evolution implies fitness of newly evolved organisms that have inherent adaptive traits because of mutations in genes. However, most mutations are detrimental, and they spoil the organism’s life, its survival and its ability to leave progeny. Some genes are extremely vital for an organism, and therefore, they tend to save their structure and do not mutate or do it very composedly. That is the case of the gene encoding PNPLA6 lysophospholipase domain that evolved in bacteria, and evolution obliged it to save its function in higher animals. In mammals, complete dysfunction of such a gene is lethal because of its high importance in placenta for early embryo development. Why is it conserved in other species, for instance insects, that have no placenta? Here we studied the role of the PNPLA6-encoding gene named swiss cheese in Drosophila melanogaster fitness. We have found that its dysfunction results in premature death of specimens and their inability to leave enough progeny. Thus, we provide the first evidence for significance of the gene that encodes the lysophospholipase enzyme in fitness of insects.Drosophila melanogaster is one of the most famous insects in biological research. It is widely used to analyse functions of different genes. The phosphatidylcholine lysophospholipase gene swiss cheese was initially shown to be important in the fruit fly nervous system. However, the role of this gene in non-nervous cell types has not been elucidated yet, and the evolutional explanation for the conservation of its function remains elusive. In this study, we analyse expression pattern and some aspects of the role of the swiss cheese gene in the fitness of Drosophila melanogaster. We describe the spatiotemporal expression of swiss cheese throughout the fly development and analyse the survival and productivity of swiss cheese mutants. We found swiss cheese to be expressed in salivary glands, midgut, Malpighian tubes, adipocytes, and male reproductive system. Dysfunction of swiss cheese results in severe pupae and imago lethality and decline of fertility, which is impressive in males. The latter is accompanied with abnormalities of male locomotor activity and courtship behaviour, accumulation of lipid droplets in testis cyst cells and decrease in spermatozoa motility. These results suggest that normal swiss cheese is important for Drosophila melanogaster fitness due to its necessity for both specimen survival and their reproductive success.

Activation of TAF9 via Danshensu-Induced Upregulation of HDAC1 Expression Alleviates Non-alcoholic Fatty Liver Disease.

Fatty acid β-oxidation is an essential pathogenic mechanism in nonalcoholic fatty liver disease (NAFLD), and TATA-box binding protein associated factor 9 (TAF9) has been reported to be involved in the regulation of fatty acid β-oxidation. However, the function of TAF9 in NAFLD, as well as the mechanism by which TAF9 is regulated, remains unclear. In this study, we aimed to investigate the signaling mechanism underlying the involvement of TAF9 in NAFLD and the protective effect of the natural phenolic compound Danshensu (DSS) against NAFLD via the HDAC1/TAF9 pathway. An in vivo model of high-fat diet (HFD)-induced NAFLD and a palmitic acid (PA)-treated AML-12 cell model were developed. Pharmacological treatment with DSS significantly increased fatty acid β-oxidation and reduced lipid droplet (LD) accumulation in NAFLD. TAF9 overexpression had the same effects on these processes both in vivo and in vitro. Interestingly, the protective effect of DSS was markedly blocked by TAF9 knockdown. Mechanistically, TAF9 was shown to be deacetylated by HDAC1, which regulates the capacity of TAF9 to mediate fatty acid β-oxidation and LD accumulation during NAFLD. In conclusion, TAF9 is a key regulator in the treatment of NAFLD that acts by increasing fatty acid β-oxidation and reducing LD accumulation, and DSS confers protection against NAFLD through the HDAC1/TAF9 pathway.