PKR is not obligatory for high-fat diet-induced obesity and its associated metabolic and inflammatory complications

Protein kinase R (PKR) has previously been suggested to mediate many of the deleterious consequences of a high-fat diet (HFD). However, previous studies have observed substantial phenotypic variability when examining the metabolic consequences of PKR deletion. Accordingly, herein, we have re-examined the role of PKR in the development of obesity and its associated metabolic complications in vivo as well as its putative lipid-sensing role in vitro. Here we show that the deletion of PKR does not affect HFD-induced obesity, hepatic steatosis or glucose metabolism, and only modestly affects adipose tissue inflammation. Treatment with the saturated fatty acid palmitate in vitro induced comparable levels of inflammation in WT and PKR KO macrophages, demonstrating that PKR is not necessary for the sensing of pro-inflammatory lipids. These results challenge the proposed role for PKR in obesity, its associated metabolic complications and its role in lipid-induced inflammation

A new emulated Monte Carlo radiative transfer disc-wind model: X-Ray Accretion Disc-wind Emulator - XRADE

We present a new X-Ray Accretion Disc-wind Emulator (xrade) based on the 2.5D Monte Carlo radiative transfer code that provides a physically motivated, self-consistent treatment of both absorption and emission from a disc wind by computing the local ionization state and velocity field within the flow. xrade is then implemented through a process that combines X-ray tracing with supervised machine learning. We develop a novel emulation method consisting in training, validating, and testing the simulated disc-wind spectra into a purposely built artificial neural network. The trained emulator can generate a single synthetic spectrum for a particular parameter set in a fraction of a second, in contrast to the few hours required by a standard Monte Carlo radiative transfer pipeline. The emulator does not suffer from interpolation issues with multidimensional spaces that are typically faced by traditional X-ray fitting packages such as xspec. xrade will be suitable to a wide number of sources across the black hole mass, ionizing luminosity, and accretion rate scales. As an example, we demonstrate the applicability of xrade to the physical interpretation of the X-ray spectra of the bright quasar PDS 456, which hosts the best-established accretion disc wind observed to date. We anticipate that our emulation method will be an indispensable tool for the development of high-resolution theoretical models, with the necessary flexibility to be optimized for the next generation microcalorimeters onboard future missions, like X-Ray Imaging and Spectroscopy Mission (XRISM)/Resolve and Athena/X-ray Integral Field Unit (X-IFU). This tool can also be implemented across a wide variety of X-ray spectral models and beyond

The first broad-band X-ray view of the narrow-line Seyfert 1 Ton S180

We present joint \textit{XMM-Newton} and \textit{NuSTAR} observations of the `bare' narrow line Seyfert 1 Ton S180 ($z=0.062$), carried out in 2016 and providing the first hard X-ray view of this luminous galaxy. We find that the 0.4--30 keV band cannot be self-consistently reproduced by relativistic reflection models, which fail to account simultaneously for the soft and hard X-ray emission. The smooth soft excess prefers extreme blurring parameters, confirmed by the nearly featureless nature of the RGS spectrum, while the moderately broad Fe K line and the modest hard excess above 10 keV appear to arise in a milder gravity regime. By allowing a different origin of the soft excess, the broadband X-ray spectrum and overall spectral energy distribution (SED) are well explained by a combination of: (a) direct thermal emission from the accretion disc, dominating from the optical to the far/extreme UV; (b) Comptonization of seed disc photons by a warm ($kT_{\rm e}\sim0.3$ keV) and optically thick ($\tau\sim10$) corona, mostly contributing to the soft X-rays; (c) Comptonization by a standard hot ($kT_{\rm e} \gtrsim 100$ keV) and optically thin ($\tau<0.5$) corona, responsible for the primary X-ray continuum; and (d) reflection from the mid/outer part of the disc. The two coronae are suggested to be rather compact, with $R_{\rm hot} \lesssim R_{\rm warm} \lesssim 10$ R$_{\rm g}$. Our SED analysis implies that Ton S180 accretes at super-Eddington rates. This is a key condition for the launch of a wind, marginal (i.e., 3.1$\sigma$ significance) evidence of which is indeed found in the RGS spectrum.Comment: 20 pages, 12 figures. Accepted for publication MNRA

Aberrant lipid metabolism disrupts calcium homeostasis causing liver endoplasmic reticulum stress in obesity.

The endoplasmic reticulum (ER) is the main site of protein and lipid synthesis, membrane biogenesis, xenobiotic detoxification and cellular calcium storage, and perturbation of ER homeostasis leads to stress and the activation of the unfolded protein response. Chronic activation of ER stress has been shown to have an important role in the development of insulin resistance and diabetes in obesity. However, the mechanisms that lead to chronic ER stress in a metabolic context in general, and in obesity in particular, are not understood. Here we comparatively examined the proteomic and lipidomic landscape of hepatic ER purified from lean and obese mice to explore the mechanisms of chronic ER stress in obesity. We found suppression of protein but stimulation of lipid synthesis in the obese ER without significant alterations in chaperone content. Alterations in ER fatty acid and lipid composition result in the inhibition of sarco/endoplasmic reticulum calcium ATPase (SERCA) activity and ER stress. Correcting the obesity-induced alteration of ER phospholipid composition or hepatic Serca overexpression in vivo both reduced chronic ER stress and improved glucose homeostasis. Hence, we established that abnormal lipid and calcium metabolism are important contributors to hepatic ER stress in obesity

A new emulated Monte Carlo radiative transfer disc-wind model: X-Ray Accretion Disc-wind Emulator – XRADE

Abstract We present a new X-Ray Accretion Disk-wind Emulator (xrade) based on the 2.5D Monte Carlo radiative transfer code which provides a physically-motivated, self-consistent treatment of both absorption and emission from a disk-wind by computing the local ionization state and velocity field within the flow. xrade is then implemented through a process that combines X-ray tracing with supervised machine learning. We develop a novel emulation method consisting in training, validating, and testing the simulated disk-wind spectra into a purposely built artificial neural network. The trained emulator can generate a single synthetic spectrum for a particular parameter set in a fraction of a second, in contrast to the few hours required by a standard Monte Carlo radiative transfer pipeline. The emulator does not suffer from interpolation issues with multi-dimensional spaces that are typically faced by traditional X-ray fitting packages such as xspec. xrade will be suitable to a wide number of sources across the black-hole mass, ionizing luminosity, and accretion rate scales. As an example, we demonstrate the applicability of xrade to the physical interpretation of the X-ray spectra of the bright quasar PDS 456, which hosts the best-established accretion-disk wind observed to date. We anticipate that our emulation method will be an indispensable tool for the development of high-resolution theoretical models, with the necessary flexibility to be optimized for the next generation micro-calorimeters on board future missions, like XRISM/Resolve and Athena/X-IFU. This tool can also be implemented across a wide variety of X-ray spectral models and beyond

Purification and In Situ Immobilization of Papain with Aqueous Two-Phase System

Papain was purified from spray-dried Carica papaya latex using aqueous two-phase system (ATPS). Then it was recovered from PEG phase by in situ immobilization or preparing cross-linked enzyme aggregates (CLEAs). The Plackett-Burman design and the central composite design (CCD) together with the response surface methodology (RSM) were used to optimize the APTS processes. The highly purified papain (96–100%) was achieved under the optimized conditions: 40% (w/w) 15 mg/ml enzyme solution, 14.33–17.65% (w/w) PEG 6000, 14.27–14.42% (w/w) NaH2PO4/K2HPO4 and pH 5.77–6.30 at 20°C. An in situ enzyme immobilization approach, carried out by directly dispersing aminated supports and chitosan beads into the PEG phase, was investigated to recover papain, in which a high immobilization yield (>90%) and activity recovery (>40%) was obtained. Moreover, CLEAs were successfully used in recovering papain from PEG phase with a hydrolytic activity hundreds times higher than the carrier-bound immobilized papain

Endoplasmic reticulum stress inhibition protects steatotic and non-steatotic livers in partial hepatectomy under ischemia–reperfusion

During partial hepatectomy, ischemia–reperfusion (I/R) is commonly applied in clinical practice to reduce blood flow. Steatotic livers show impaired regenerative response and reduced tolerance to hepatic injury. We examined the effects of tauroursodeoxycholic acid (TUDCA) and 4-phenyl butyric acid (PBA) in steatotic and non-steatotic livers during partial hepatectomy under I/R (PH+I/R). Their effects on the induction of unfolded protein response (UPR) and endoplasmic reticulum (ER) stress were also evaluated. We report that PBA, and especially TUDCA, reduced inflammation, apoptosis and necrosis, and improved liver regeneration in both liver types. Both compounds, especially TUDCA, protected both liver types against ER damage, as they reduced the activation of two of the three pathways of UPR (namely inositol-requiring enzyme and PKR-like ER kinase) and their target molecules caspase 12, c-Jun N-terminal kinase and C/EBP homologous protein-10. Only TUDCA, possibly mediated by extracellular signal-regulated kinase upregulation, inactivated glycogen synthase kinase-3β. This is turn, inactivated mitochondrial voltage-dependent anion channel, reduced cytochrome c release from the mitochondria and caspase 9 activation and protected both liver types against mitochondrial damage. These findings indicate that chemical chaperones, especially TUDCA, could protect steatotic and non-steatotic livers against injury and regeneration failure after PH+I/R

Rhaponticum acaule (L) DC essential oil: chemical composition, in vitro antioxidant and enzyme inhibition properties

Background: α-glucosidase is a therapeutic target for diabetes mellitus (DM) and α-glucosidase inhibitors play a vital role in the treatments for the disease. Furthermore, xanthine oxidase (XO) is a key enzyme that catalyzes hypoxanthine and xanthine to uric acid which at high levels can lead to hyperuricemia which is an important cause of gout. Pancreatic lipase (PL) secreted into the duodenum plays a key role in the digestion and absorption of fats. For its importance in lipid digestion, PL represents an attractive target for obesity prevention. Methods: The flowers essential oil of Rhaponticum acaule (L) DC (R. acaule) was characterized using gas chromatography-mass spectrometry (GC-MS). The antioxidant activities of R. acaule essential oil (RaEO) were also determined using 2,2’-azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS), reducing power, phosphomolybdenum, and DNA nicking assays. The inhibitory power of RaEO against α-glucosidase, xanthine oxidase and pancreatic lipase was evaluated. Enzyme kinetic studies using Michaelis-Menten and the derived Lineweaver-Burk (LB) plots were performed to understand the possible mechanism of inhibition exercised by the components of this essential oil. Results: The result revealed the presence of 26 compounds (97.4%). The main constituents include germacrene D (49.2%), methyl eugenol (8.3%), (E)-β-ionone (6.2%), β-caryophyllene (5.7%), (E,E)-α-farnesene (4.2%), bicyclogermacrene (4.1%) and (Z)-α-bisabolene (3.7%). The kinetic inhibition study showed that the essential oil demonstrated a strong α-glucosidase inhibiton and it was a mixed inhibitor. On the other hand, our results evidenced that this oil exhibited important xanthine oxidase inhibitory effect, behaving as a non-competitive inhibitor. The essential oil inhibited the turkey pancreatic lipase, with maximum inhibition of 80% achieved at 2 mg/mL. Furthermore, the inhibition of turkey pancreatic lipase by RaEO was an irreversible one. Conclusion: The results revealed that the RaEO is a new promising potential source of antioxidant compounds, endowed with good practical applications for human health. Keywords: α-glucosidase, Antioxidant activity, Chemical composition, Pancreatic lipase inhibition, Rhaponticum acaule essential oil, Xanthine oxidase

Differing Endoplasmic Reticulum Stress Response to Excess Lipogenesis versus Lipid Oversupply in Relation to Hepatic Steatosis and Insulin Resistance

Mitochondrial dysfunction and endoplasmic reticulum (ER) stress have been implicated in hepatic steatosis and insulin resistance. The present study investigated their roles in the development of hepatic steatosis and insulin resistance during de novo lipogenesis (DNL) compared to extrahepatic lipid oversupply. Male C57BL/6J mice were fed either a high fructose (HFru) or high fat (HFat) diet to induce DNL or lipid oversupply in/to the liver. Both HFru and HFat feeding increased hepatic triglyceride within 3 days (by 3.5 and 2.4 fold) and the steatosis remained persistent from 1 week onwards (p<0.01 vs Con). Glucose intolerance (iAUC increased by ∼60%) and blunted insulin-stimulated hepatic Akt and GSK3β phosphorylation (∼40–60%) were found in both feeding conditions (p<0.01 vs Con, assessed after 1 week). No impairment of mitochondrial function was found (oxidation capacity, expression of PGC1α, CPT1, respiratory complexes, enzymatic activity of citrate synthase & β-HAD). As expected, DNL was increased (∼60%) in HFru-fed mice and decreased (32%) in HFat-fed mice (all p<0.05). Interestingly, associated with the upregulated lipogenic enzymes (ACC, FAS and SCD1), two (PERK/eIF2α and IRE1/XBP1) of three ER stress pathways were significantly activated in HFru-fed mice. However, no significant ER stress was observed in HFat-fed mice during the development of hepatic steatosis. Our findings indicate that HFru and HFat diets can result in hepatic steatosis and insulin resistance without obvious mitochondrial defects via different lipid metabolic pathways. The fact that ER stress is apparent only with HFru feeding suggests that ER stress is involved in DNL per se rather than resulting from hepatic steatosis or insulin resistance