O-GlcNAc modification in endothelial cells modulates adiposity via fat absorption from the intestine in mice.

Introduction: Endothelial cells have a crucial function in transporting and exchanging various nutrients. O-GlcNAcylation, mediated by O-GlcNAc transferase (OGT), involves the addition of N-acetylglucosamine to proteins and serves as an intracellular nutrient sensing mechanism. However, the role of O-GlcNAcylation in endothelial cells remains poorly understood. Objective: This study investigated the role of O-GlcNAcylation in endothelial cells. Methods: Endothelial-cell-specific Ogt -knockout mice (Ogt-ECKO) were generated by crossing Ogt-floxed mice (Ogt-flox) with VE-Cadherin Cre ERT2 mice. Ogt-ECKO mice and Ogt-flox control mice were subjected to a normal or high-fat diet, and their body weight, glucose metabolism, and lipid metabolism were examined. Results: Ogt-ECKO mice exhibited reduced body weight compared with Ogt-flox control mice under a high-fat diet. Lipid absorption was significantly impaired in Ogt-ECKO mice. Changes in the intercellular junctions of small intestinal lacteal endothelial cells from a button-like to a zipper-like configuration were observed. Furthermore, Ogt-ECKO mice showed decreased expression of VEGFR3. The administration of a nitric oxide donor restored lipid absorption and reversed the morphological alterations in Ogt-ECKO mice. Conclusions: These findings demonstrate the critical role of O-GlcNAcylation in endothelial cells in lipid absorption in the intestine through the modulation of lacteal junction morphology. These results provide novel insight into the metabolic regulatory mechanisms under physiological conditions and have implications for the development of new therapeutic strategies for obesity.

Cytosolic calcium regulates hepatic mitochondrial oxidation, intrahepatic lipolysis, and gluconeogenesis via CAMKII activation.

To examine the roles of mitochondrial calcium Ca2+ ([Ca2+]mt) and cytosolic Ca2+ ([Ca2+]cyt) in the regulation of hepatic mitochondrial fat oxidation, we studied a liver-specific mitochondrial calcium uniporter knockout (MCU KO) mouse model with reduced [Ca2+]mt and increased [Ca2+]cyt content. Despite decreased [Ca2+]mt, deletion of hepatic MCU increased rates of isocitrate dehydrogenase flux, α-ketoglutarate dehydrogenase flux, and succinate dehydrogenase flux in vivo. Rates of [14C16]palmitate oxidation and intrahepatic lipolysis were increased in MCU KO liver slices, which led to decreased hepatic triacylglycerol content. These effects were recapitulated with activation of CAMKII and abrogated with CAMKII knockdown, demonstrating that [Ca2+]cyt activation of CAMKII may be the primary mechanism by which MCU deletion promotes increased hepatic mitochondrial oxidation. Together, these data demonstrate that hepatic mitochondrial oxidation can be dissociated from [Ca2+]mt and reveal a key role for [Ca2+]cyt in the regulation of hepatic fat mitochondrial oxidation, intrahepatic lipolysis, gluconeogenesis, and lipid accumulation.

Small molecule inhibition of glycogen synthase I reduces muscle glycogen content and improves biomarkers in a mouse model of Pompe disease.

Pompe disease is a rare genetic disorder caused by a deficiency of the enzyme acid alpha-glucosidase (GAA). This enzyme is responsible for breaking down glycogen, leading to the abnormal accumulation of glycogen, which results in progressive muscle weakness and metabolic dysregulation. In this study, we investigated the hypothesis that the small molecule inhibition of glycogen synthase I (GYS1) may reduce muscle glycogen content and improve metabolic dysregulation in a mouse model of Pompe disease. To address this hypothesis, we studied four groups of male mice: a control group of wild-type B6129SF1/J mice fed either regular chow (WT) or a GYS1 inhibitor (MZ-101) diet (WT-GYS1), and Pompe model mice B6;129-Gaatm1Rabn/J fed either regular chow (GAA-KO) or MZ-101 diet (GAA-GYS1) for 7 days. Our findings revealed that GAA-KO mice exhibited abnormal glycogen accumulation in the gastrocnemius, heart, and diaphragm. In contrast, inhibiting GYS1 reduced glycogen levels in all tissues compared to GAA-KO mice. Furthermore, GAA-KO mice displayed reduced spontaneous activity during the dark cycle compared to WT mice, while GYS1 inhibition counteracted this effect. Compared to GAA-KO mice, GAA-GYS1 mice exhibited improved glucose tolerance and whole-body insulin sensitivity. These improvements in insulin sensitivity could be attributed to increased AMPK phosphorylation in the gastrocnemius of WT-GYS1 and GAA-GYS1 mice. Additionally, the GYS1 inhibitor led to a reduction in the phosphorylation of GSS641 and the LC3 autophagy marker. Together, our results suggest that targeting GYS1 could serve as a potential strategy for treating glycogen storage disorders and metabolic dysregulation.

Intrathecal dexamethasone-bupivacaine combination with bupivacaine alone in spinal anesthesia for cesarean delivery.

Background: Postoperative pain management can be achieved by adjuvant medications during the analgesia procedure. The study investigated the effect of intrathecal dexamethasone-bupivacaine combination with bupivacaine alone in spinal anesthesia for cesarean delivery. Methods: This randomized, double-blind clinical examination included 50 females who had previously experienced a cesarean section. The participants were assigned randomly into two categories: the intervention group, received intrathecal bupivacaine-dexamethasone, and the control group, received intrathecal bupivacaine-normal saline. Levels of pain were evaluated using a 10 cm visual analog scale (VAS) at intervals of 30 minutes, 1 hour, 2 hours after the operation. The span of the sensory block and postoperative analgesia were assessed. Results: The inclusion of intrathecal dexamethasone with bupivacaine resulted in a significant enhancement in the duration of pain relief during the intervention, lasting for an average of 473.4 ± 39.95 minutes (p<0.001). The duration of sensory and motor block analgesia in the intervention group was more than the control group (128.32 ± 7.30 vs. 92.84 ± 7.84) and (155.6±12.34 vs. 126.16±11.89), respectively (p<0.001). Pain score on the VAS scale at 30, 60, and 120 minutes was significantly lower in the intervention group (p<0.001). There was no difference in side effects and onset time between the study groups. Conclusion: The inclusion of intrathecal dexamethasone alongside bupivacaine has demonstrated enhancement in the duration of sensory block during spinal anesthesia. This improvement was observed without any alterations in the time it takes for the anesthesia to take effect and without any adverse effects during the postoperative period.

Latrophilin-2 mediates fluid shear stress mechanotransduction at endothelial junctions.

Endothelial cell responses to fluid shear stress from blood flow are crucial for vascular development, function and disease. A complex of PECAM-1, VE-cadherin, VEGF receptors (VEGFRs) and PlexinD1 located at cell-cell junctions mediates many of these events. But available evidence suggests that another mechanosensor upstream of PECAM-1 initiates signaling. Hypothesizing that GPCR and Gα proteins may serve this role, we performed siRNA screening of Gα subunits and found that Gαi2 and Gαq/11 are required for activation of the junctional complex. We then developed a new activation assay, which showed that these G proteins are activated by flow. We next mapped the Gα residues required for activation and developed an affinity purification method that used this information to identify latrophilin-2 (Lphn-2/ADGRL2) as the upstream GPCR. Latrophilin-2 is required for all PECAM-1 downstream events tested. In both mice and zebrafish, latrophilin-2 is required for flow-dependent angiogenesis and artery remodeling. Furthermore, endothelial specific knockout demonstrates that latrophilin plays a role in flow-dependent artery remodeling. Human genetic data reveal a correlation between the latrophilin-2-encoding Adgrl2 gene and cardiovascular disease. Together, these results define a pathway that connects latrophilin-dependent G protein activation to subsequent endothelial signaling, vascular physiology and disease.

Latrophilin-2 mediates fluid shear stress mechanotransduction at endothelial junctions.

Endothelial cell responses to fluid shear stress from blood flow are crucial for vascular development, function, and disease. A complex of PECAM-1, VE-cadherin, VEGF receptors (VEGFRs), and Plexin D1 located at cell-cell junctions mediates many of these events. However, available evidence suggests that another mechanosensor upstream of PECAM-1 initiates signaling. Hypothesizing that GPCR and Gα proteins may serve this role, we performed siRNA screening of Gα subunits and found that Gαi2 and Gαq/11 are required for activation of the junctional complex. We then developed a new activation assay, which showed that these G proteins are activated by flow. We next mapped the Gα residues required for activation and developed an affinity purification method that used this information to identify latrophilin-2 (Lphn2/ADGRL2) as the upstream GPCR. Latrophilin-2 is required for all PECAM-1 downstream events tested. In both mice and zebrafish, latrophilin-2 is required for flow-dependent angiogenesis and artery remodeling. Furthermore, endothelial-specific knockout demonstrates that latrophilin plays a role in flow-dependent artery remodeling. Human genetic data reveal a correlation between the latrophilin-2-encoding Adgrl2 gene and cardiovascular disease. Together, these results define a pathway that connects latrophilin-dependent G protein activation to subsequent endothelial signaling, vascular physiology, and disease.

Corrosion response of steels fabricated through arc directed energy deposition additive manufacturing: a review.

Steels exhibit distinct properties that underscore their pivotal role in critical industries, such as maritime, aerospace, automotive, petrochemical, and biomedicine. In recent times, there has been an increasing trend towards manufacturing near-net-shape steel components through various additive manufacturing (AM) modalities, utilizing intricate 3D model data. Initially, powder bed fusion (PBF) technology garnered significant attention for the fabrication of steel components. Nonetheless, arc-directed energy deposition (arc-DED), also known as wire arc additive manufacturing (WAAM) technology, is progressively gaining prominence in the AM enterprise due to its high production rate, the ability to print large-scale components, and notably, reduced capital investment. While early research on WAAM-fabricated steels primarily focused on microstructural and mechanical characteristics, there is an increasing emphasis on the corrosion performance of WAAM steel components. These components often encounter exposure to corrosive environments in their intended applications. The existing literature lacks a comprehensive review that delves into the nuanced factors influencing the corrosion behavior of WAAM-fabricated steels and the primary corrosion mechanisms governing their degradation. Therefore, this review is dedicated to exploring the corrosion properties of WAAM-fabricated steels, identifying key parameters influencing their degradation behavior. Moreover, it offers an in-depth examination and discussion of the underlying mechanisms governing corrosion-induced deterioration. Furthermore, this review meticulously scrutinizes the microstructural features and WAAM technologies, providing clarity and organization regarding details relevant to the corrosion of WAAM steel components. To conclude, the paper highlights the existing research gaps related to the corrosion of WAAM steel, delineating potential avenues for future research.

Effect of Surface Nanocrystallization on Wear Behavior of Steels: A Review.

Ferrous alloys, particularly steels, form a specialized class of metallic materials extensively employed in industrial sectors to combat deterioration and failures caused by wear. Despite their commendable mechanical properties, steels are not immune to wear-induced degradation. In this context, surface nanocrystallization (SNC) technologies have carved a distinct niche for themselves by enabling the nanostructuring of the surface layer (with grain sizes < 100 nm). This process enhances overall mechanical properties to a level desirable for wear resistance while preserving the chemical composition. Existing literature has consistently highlighted the efficacy of various SNC methods in improving the wear resistance of ferrous alloys, positioning SNC as a promising tool to extend materials' service life in practical applications. This review provides a comprehensive examination of the SNC techniques employed in surface treatment of ferrous alloys and their impact on wear behavior. We delved into the underlying mechanisms governing wear in SNC-treated Fe-based alloys and concluded with a discussion on current challenges and future perspectives in this evolving field.

A review of the corrosion behavior of conventional and additively manufactured nickel-aluminum bronze (NAB) alloys: current status and future challenges.

The growing demand for materials with exceptional corrosion resistance and mechanical properties in the aerospace and ocean industries has led to increased research interest in versatile alloys like nickel-aluminum bronze (NAB). NABs exhibit excellent corrosion performance due to the formation of a protective, duplex corrosion product film on the surface, which is largely influenced by their complex microstructure. While NABs are typically produced as cast or wrought products, the emergence of additive manufacturing (AM) technologies has enabled 3D printing of near-net-shape NABs with intricate geometries. This paper provides a critical review of the corrosion properties, passivity, and microstructural characteristics of conventionally produced and AMed NAB alloys, as well as the fundamental mechanisms governing their corrosion behavior under varying conditions. Additionally, it highlights the current research gap and unprecedented challenges associated with the corrosion behavior of traditional and AMed NABs.

MAD2-Dependent Insulin Receptor Endocytosis Regulates Metabolic Homeostasis.

Insulin activates insulin receptor (IR) signaling and subsequently triggers IR endocytosis to attenuate signaling. Cell division regulators MAD2, BUBR1, and p31comet promote IR endocytosis on insulin stimulation. Here, we show that genetic ablation of the IR-MAD2 interaction in mice delays IR endocytosis, increases IR levels, and prolongs insulin action at the cell surface. This in turn causes a defect in insulin clearance and increases circulating insulin levels, unexpectedly increasing glucagon levels, which alters glucose metabolism modestly. Disruption of the IR-MAD2 interaction increases serum fatty acid concentrations and hepatic fat accumulation in fasted male mice. Furthermore, disruption of the IR-MAD2 interaction distinctly changes metabolic and transcriptomic profiles in the liver and adipose tissues. Our findings establish the function of cell division regulators in insulin signaling and provide insights into the metabolic functions of IR endocytosis. ARTICLE HIGHLIGHTS: The physiological role of IR endocytosis in insulin sensitivity remains unclear. Disruption of the IR-MAD2 interaction delays IR endocytosis and prolongs insulin signaling. IR-MAD2 controls insulin clearance and glucose metabolism. IR-MAD2 maintains energy homeostasis.

The Prophylaxis Effect of Ephedrine on Hemodynamic Variation in Patients Undergoing Percutaneous Nephrolithotomy Surgery with Spinal Anesthesia.

Background: Performing spinal anesthesia with at least hemodynamic variation and complications is always challenging for anesthesiologists. In this study, we investigated the effect of ephedrine and placebo on hemodynamic changes in patients undergoing percutaneous nephrolithotomy with spinal anesthesia. Methods: This randomized, double-blind prospective clinical trial was conducted on 120 patients aged 20‒60 years with ASA (American Society of Anesthesiologists) classes I and II. Patients who were candidates for percutaneous nephrolithotomy with spinal anesthesia were divided into intervention (received 1 cc = 5 mg ephedrine) and control groups (received 1 cc normal saline). All vital parameters, including HR (heart rate) and NIBP (noninvasive blood pressure), were recorded perioperatively T0-T25) and finally at the end of surgery time (Tf). The results were analyzed by SPSS software version 23, and a P value ≤0.05 was considered significant. Results: The mean arterial pressure during surgery between T3 and T9 and the mean heart rate in times of T3-T8 in the intervention group were higher than in the control group, and this difference was statistically significant (P < 0.05). The incidence of hypotension, bradycardia, nausea, and vomiting and the amount of prescribed ephedrine, atropine, and ondansetron in the control group were higher than in the intervention group (P=0.001). Seven patients in the control group and four in the intervention group had shivering, but this difference was not statistically significant (P=0.43). Conclusion: This study showed the effectiveness of the prescription of 5 mg ephedrine two minutes before changing from the lithotomy position to the supine in maintaining hemodynamic stability, reducing hypotension, bradycardia, nausea, and vomiting, and the amount of prescribed ephedrine, atropine, and ondansetron. Trial Registrations. This trial is registered with IRCT20160430027677N22.

Inhibition of HSD17B13 protects against liver fibrosis by inhibition of pyrimidine catabolism in nonalcoholic steatohepatitis.

Nonalcoholic fatty liver disease (NAFLD) is the most common chronic liver disease, in which prognosis is determined by liver fibrosis. A common variant in hydroxysteroid 17-beta dehydrogenase 13 (HSD17B13, rs72613567-A) is associated with a reduced risk of fibrosis in NAFLD, but the underlying mechanism(s) remains unclear. We investigated the effects of this variant in the human liver and in Hsd17b13 knockdown in mice by using a state-of-the-art metabolomics approach. We demonstrate that protection against liver fibrosis conferred by the HSD17B13 rs72613567-A variant in humans and by the Hsd17b13 knockdown in mice is associated with decreased pyrimidine catabolism at the level of dihydropyrimidine dehydrogenase. Furthermore, we show that hepatic pyrimidines are depleted in two distinct mouse models of NAFLD and that inhibition of pyrimidine catabolism by gimeracil phenocopies the HSD17B13-induced protection against liver fibrosis. Our data suggest pyrimidine catabolism as a therapeutic target against the development of liver fibrosis in NAFLD.

Distinct subcellular localisation of intramyocellular lipids and reduced PKCε/PKCθ activity preserve muscle insulin sensitivity in exercise-trained mice.

AIMS/HYPOTHESIS: Athletes exhibit increased muscle insulin sensitivity, despite increased intramuscular triacylglycerol content. This phenomenon has been coined the 'athlete's paradox' and is poorly understood. Recent findings suggest that the subcellular distribution of sn-1,2-diacylglycerols (DAGs) in the plasma membrane leading to activation of novel protein kinase Cs (PKCs) is a crucial pathway to inducing insulin resistance. Here, we hypothesised that regular aerobic exercise would preserve muscle insulin sensitivity by preventing increases in plasma membrane sn-1,2-DAGs and activation of PKCε and PKCθ despite promoting increases in muscle triacylglycerol content. METHODS: C57BL/6J mice were allocated to three groups (regular chow feeding [RC]; high-fat diet feeding [HFD]; RC feeding and running wheel exercise [RC-EXE]). We used a novel LC-MS/MS/cellular fractionation method to assess DAG stereoisomers in five subcellular compartments (plasma membrane [PM], endoplasmic reticulum, mitochondria, lipid droplets and cytosol) in the skeletal muscle. RESULTS: We found that the HFD group had a greater content of sn-DAGs and ceramides in multiple subcellular compartments compared with the RC mice, which was associated with an increase in PKCε and PKCθ translocation. However, the RC-EXE mice showed, of particular note, a reduction in PM sn-1,2-DAG and ceramide content when compared with HFD mice. Consistent with the PM sn-1,2-DAG-novel PKC hypothesis, we observed an increase in phosphorylation of threonine1150 on the insulin receptor kinase (IRKT1150), and reductions in insulin-stimulated IRKY1162 phosphorylation and IRS-1-associated phosphoinositide 3-kinase activity in HFD compared with RC and RC-EXE mice, which are sites of PKCε and PKCθ action, respectively. CONCLUSIONS/INTERPRETATION: These results demonstrate that lower PKCθ/PKCε activity and sn-1,2-DAG content, especially in the PM compartment, can explain the preserved muscle insulin sensitivity in RC-EXE mice.

Sex- and strain-specific effects of mitochondrial uncoupling on age-related metabolic diseases in high-fat diet-fed mice.

Mild uncoupling of oxidative phosphorylation is an intrinsic property of all mitochondria and may have evolved to protect cells against the production of damaging reactive oxygen species. Therefore, compounds that enhance mitochondrial uncoupling are potentially attractive anti-aging therapies; however, chronic ingestion is associated with a number of unwanted side effects. We have previously developed a controlled-release mitochondrial protonophore (CRMP) that is functionally liver-directed and promotes oxidation of hepatic triglycerides by causing a subtle sustained increase in hepatic mitochondrial inefficiency. Here, we sought to leverage the higher therapeutic index of CRMP to test whether mild mitochondrial uncoupling in a liver-directed fashion could reduce oxidative damage and improve age-related metabolic disease and lifespan in diet-induced obese mice. Oral administration of CRMP (20 mg/[kg-day] × 4 weeks) reduced hepatic lipid content, protein kinase C epsilon activation, and hepatic insulin resistance in aged (74-week-old) high-fat diet (HFD)-fed C57BL/6J male mice, independently of changes in body weight, whole-body energy expenditure, food intake, or markers of hepatic mitochondrial biogenesis. CRMP treatment was also associated with a significant reduction in hepatic lipid peroxidation, protein carbonylation, and inflammation. Importantly, long-term (49 weeks) hepatic mitochondrial uncoupling initiated late in life (94-104 weeks), in conjugation with HFD feeding, protected mice against neoplastic disorders, including hepatocellular carcinoma (HCC), in a strain and sex-specific manner. Taken together, these studies illustrate the complex variation of aging and provide important proof-of-concept data to support further studies investigating the use of liver-directed mitochondrial uncouplers to promote healthy aging in humans.

Effects of Secondary-Phase Formation on the Electrochemical Performance of a Wire Arc Additive Manufactured 420 Martensitic Stainless Steel under Different Heat Treatment Conditions.

This study aims to investigate the effects of annealing, quenching, and tempering (Q&T) heat treatments on the microstructure, crystallographic orientation, and electrochemical performance of a wall shaped 420 martensitic stainless steel part fabricated by wire arc additive manufacturing technology. The formation of a martensitic matrix with delta ferrite in the as-printed sample, islands of spherical chromium carbides embedded in a ferritic matrix in annealed sample, and intergranular chromium-rich carbides along the primary austenite grain boundaries in addition to intra-lath Fe-rich carbides in the quenching and tempering heat treated sample were detected. To characterize the corrosion performance of the fabricated samples, open circuit potential, potentiodynamic polarization, and electrochemical impedance spectroscopy tests were performed on all samples in aerated 3.5 wt.% NaCl electrolyte at room temperature. The corrosion morphology of the as-printed sample was characterized by localized corrosion attacks adjacent to the delta ferrite phase, while severe pitting occurred in the annealed sample due to the high susceptibility of ferritic matrix-carbide interface to pitting. In contrast to the as-printed and annealed sample, the electrochemical performance of the quenched and subsequently tempered samples was found to be significantly improved, ascribed to elimination of the chromium depleted regions adjacent to the delta ferrite phase, and enhanced protectiveness of the passive film on the alloy's surface.

Status, role, and performance of emergency medicine specialists in Iran: A qualitative study.

INTRODUCTION: The performance of the emergency department (ED) as one of the main parts of hospitals, have a great impact on the performance of the whole-hospital. In Iran, the official education program of this discipline was started in 2001 and has expanded in most medical universities. Given the unprecedentedness of emergency medicine (EM), there are limited studies about this specialty. Thus, this study aims to explore the status, role, and performance of Iranian EM specialists. MATERIALS AND METHODS: This qualitative study was conducted using content analysis of 19 semi-structured interviews with EM specialists and key informant. Purposive sampling was conducted, and some teaching and nonstate hospitals in different geographic regions of Tehran city were selected. Conducting interviews continued until reaching the data saturation. Thematic analysis was employed. Extracted themes were reviewed and confirmed by some of the participants. RESULTS: The study results were categorized within five main themes; included the role of ED from EM specialists' viewpoint, EM specialists' viewpoint on their discipline, performance of EM specialists (including medical, managerial, and economic performance), and role of EM specialists in patient satisfaction; and opportunities and challenges of EM specialists. CONCLUSION: Overall, the study findings highlighted the effectiveness and positive medical, managerial and economic impacts of EM in Iran, inside and beyond hospitals. However, the study addressed significant opportunities some solvable challenges in educational, professional and economic domains, and interdisciplinary relationships. Further studies are recommended for comprehensive exploring viewpoint of other disciplines and stakeholders.

Analysis of the Man-Made Causes of Shiraz Flash Flood: Iran, 2019.

Flood is the most common natural hazard in Iran, which annually affects the environment and human lives. On March 25, 2019 in Shiraz-Iran, following a heavy rainfall, the occurrence of a flash flood caused an extensive number of deaths, injuries, and vehicle demolitions in a short time. Evidence suggests that man-made causes of the incident, including unsustainable urban development and lack of early warning services, have played a more influential role compared with its natural causes. This study has attempted to substantiate that understanding disaster risks, as the first priority of Sendai Framework for Disaster Risk Reduction (SFDRR) 2015-2030, directly impacts the decisions and actions of policymakers, local authorities, and the public. To provide more safety, mitigation, and disaster risk reduction, attention should primarily be paid on making a cultural paradigm shift through providing sufficient training in developing appropriate disaster risk perception in the community at large.

On the Post-Printing Heat Treatment of a Wire Arc Additively Manufactured ER70S Part.

Wire arc additive manufacturing (WAAM) is known to induce a considerable microstructural inhomogeneity and anisotropy in mechanical properties, which can potentially be minimized by adopting appropriate post-printing heat treatment. In this paper, the effects of two heat treatment cycles, including hardening and normalizing on the microstructure and mechanical properties of a WAAM-fabricated low-carbon low-alloy steel (ER70S-6) are studied. The microstructure in the melt pools of the as-printed sample was found to contain a low volume fraction of lamellar pearlite formed along the grain boundaries of polygonal ferrite as the predominant micro-constituents. The grain coarsening in the heat affected zone (HAZ) was also detected at the periphery of each melt pool boundary, leading to a noticeable microstructural inhomogeneity in the as-fabricated sample. In order to modify the nonuniformity of the microstructure, a normalizing treatment was employed to promote a homogenous microstructure with uniform grain size throughout the melt pools and HAZs. Differently, the hardening treatment contributed to the formation of two non-equilibrium micro-constituents, i.e., acicular ferrite and bainite, primarily adjacent to the lamellar pearlite phase. The results of microhardness testing revealed that the normalizing treatment slightly decreases the microhardness of the sample; however, the formation of non-equilibrium phases during hardening process significantly increased the microhardness of the component. Tensile testing of the as-printed part in the building and deposition directions revealed an anisotropic ductility. Although normalizing treatment did not contribute to the tensile strength improvement of the component, it suppressed the observed anisotropy in ductility. On the contrary, the hardening treatment raised the tensile strength, but further intensified the anisotropic behavior of the component.

Glucagon stimulates gluconeogenesis by INSP3R1-mediated hepatic lipolysis.

Although it is well-established that reductions in the ratio of insulin to glucagon in the portal vein have a major role in the dysregulation of hepatic glucose metabolism in type-2 diabetes1-3, the mechanisms by which glucagon affects hepatic glucose production and mitochondrial oxidation are poorly understood. Here we show that glucagon stimulates hepatic gluconeogenesis by increasing the activity of hepatic adipose triglyceride lipase, intrahepatic lipolysis, hepatic acetyl-CoA content and pyruvate carboxylase flux, while also increasing mitochondrial fat oxidation-all of which are mediated by stimulation of the inositol triphosphate receptor 1 (INSP3R1). In rats and mice, chronic physiological increases in plasma glucagon concentrations increased mitochondrial oxidation of fat in the liver and reversed diet-induced hepatic steatosis and insulin resistance. However, these effects of chronic glucagon treatment-reversing hepatic steatosis and glucose intolerance-were abrogated in Insp3r1 (also known as Itpr1)-knockout mice. These results provide insights into glucagon biology and suggest that INSP3R1 may represent a target for therapies that aim to reverse nonalcoholic fatty liver disease and type-2 diabetes.

Slc20a1/Pit1 and Slc20a2/Pit2 are essential for normal skeletal myofiber function and survival.

Low blood phosphate (Pi) reduces muscle function in hypophosphatemic disorders. Which Pi transporters are required and whether hormonal changes due to hypophosphatemia contribute to muscle function is unknown. To address these questions we generated a series of conditional knockout mice lacking one or both house-keeping Pi transporters Pit1 and Pit2 in skeletal muscle (sm), using the postnatally expressed human skeletal actin-cre. Simultaneous conditional deletion of both transporters caused skeletal muscle atrophy, resulting in death by postnatal day P13. smPit1-/-, smPit2-/- and three allele mutants are fertile and have normal body weights, suggesting a high degree of redundance for the two transporters in skeletal muscle. However, these mice show a gene-dose dependent reduction in running activity also seen in another hypophosphatemic model (Hyp mice). In contrast to Hyp mice, grip strength is preserved. Further evaluation of the mechanism shows reduced ERK1/2 activation and stimulation of AMP kinase in skeletal muscle from smPit1-/-; smPit2-/- mice consistent with energy-stress. Similarly, C2C12 myoblasts show a reduced oxygen consumption rate mediated by Pi transport-dependent and ERK1/2-dependent metabolic Pi sensing pathways. In conclusion, we here show that Pit1 and Pit2 are essential for normal myofiber function and survival, insights which may improve management of hypophosphatemic myopathy.