Energy Homeostasis During Fasting and Fed States: Foxa2 Regulates Hepatic Amino Acid Uptake

In MODY3 patients, as well in the experimental mouse model, the Tcf1 -/- mouse, there is a defect in arginine-induced insulin secretion. In this thesis, we examined the role of arginine transporters in the insulin secreting cells in arginine-induced insulin secretion. We first characterized arginine uptake by MIN6 cells as having a Km of 102.6 μM and being partially Na-dependent and entirely Cl-dependent. We then examined Tcf1 -/- pancreatic islets, which are defective for arginine-induced insulin secretion. Using gene expression array analysis and semi-quantitative RT-PCR analysis on pancreatic islets from Tcf1 +/+ and Tcf1 -/- mice, we found eight arginine transporters expressed in the pancreatic islets with only two transporters, mNAT3 and CAT3, regulated by Tcf1 in the islets. Using siRNA-mediated knockdown of both transporters, we found that these transporters are not required for arginine-induced insulin secretion in MIN6 cells. We conclude that there is a high level of redundancy for arginine transport into insulin-secreting cells. We also examined the regulation of mNAT3 in the liver. The liver is the main site for gluconeogenesis during fasting, and has been shown to increase amino acid uptake during periods of low nutrient intake to increase its substrate iv pool for glucose production. We found that mNAT3 is upregulated during fasting, and that this response is abolished with insulin. Using mice injected with adenovirus expressing either Foxa2 or GFP as a control, we found that Foxa2 mediates this insulin-sensitive increase in hepatic mNAT3 expression. Through electrophoretic mobility shift assays and chromatin immunoprecipitation experiments, we found that Foxa2 binds to promoter elements of the mNAT3 gene, Slc38a4. We also showed these promoter elements to be important for Foxa2 transactivation using luciferase reporter gene assays. In addition, with liver perfusion experiments using mice infected with adenovirus, we found increased arginine uptake by livers overexpressing Foxa2. Thus, we have identified Foxa2 as a mediator for increased hepatic amino acid uptake during fasting

Cell Death and Neuronal Replacement during Formation of the Avian Ciliary Ganglion

AbstractProgrammed cell death is a prominent feature of embryonic development and is essential in matching the number of neurons to the target tissues that are innervated. Although a decrease in neuronal number which coincides with peripheral synaptogenesis has been well documented in the avian ciliary ganglion, it has not been clear whether cell death also occurs earlier. We observed TUNEL-positive neurons as early as stage 24, with a large peak at stage 29. This cell death at stage 29 was followed by a statistically significant (P < 0.0001) decrease in total neuron number at stage 31. The total number of neurons was recovered by stage 33/34. This suggested that dying neurons were replaced by new neurons. This replacement process did not involve proliferation because bromodeoxyuridine applied at stages 29 and 31 was unable to label neurons harvested at stage 33/34. The peak of cell death at stage 29 was increased 2.3-fold by removal of the optic vesicle and was reduced by 50% when chCNTF was overexpressed. Taken together, these results suggest that the regulation of neuron number in the ciliary ganglion is a dynamic process involving both cell death and neural replacement from postmitotic precursors prior to differentiation and innervation of target tissues

Fibulin-4 is essential for maintaining arterial wall integrity in conduit but not muscular arteries

Homozygous or compound heterozygous mutations in fibulin-4 (FBLN4) lead to autosomal recessive cutis laxa type 1B (ARCL1B), a multisystem disorder characterized by significant cardiovascular abnormalities, including abnormal elastin assembly, arterial tortuosity, and aortic aneurysms. We sought to determine the consequences of a human disease-causing mutation in FBLN4 (E57K) on the cardiovascular system and vascular elastic fibers in a mouse model of ARCL1B. Fbln4E57K/E57K mice were hypertensive and developed arterial elongation, tortuosity, and ascending aortic aneurysms. Smooth muscle cell organization within the arterial wall of large conducting vessels was abnormal, and elastic fibers were fragmented and had a moth-eaten appearance. In contrast, vessel wall structure and elastic fiber integrity were normal in resistance/muscular arteries (renal, mesenteric, and saphenous). Elastin cross-linking and total elastin content were unchanged in large or small arteries, whereas elastic fiber architecture was abnormal in large vessels. While the E57K mutation did not affect Fbln4 mRNA levels, FBLN4 protein was lower in the ascending aorta of mutant animals compared to wild-type arteries but equivalent in mesenteric arteries. We found a differential role of FBLN4 in elastic fiber assembly, where it functions mainly in large conduit arteries. These results suggest that elastin assembly has different requirements depending on vessel type. Normal levels of elastin cross-links in mutant tissue call into question FBLN4\u27s suggested role in mediating lysyl oxidase-elastin interactions. Future studies investigating tissuespecific elastic fiber assembly may lead to novel therapeutic interventions for ARCL1B and other disorders of elastic fiber assembly. 2017 © The Authors, some rights reserved

Loss of Sphingosine Kinase 1/S1P Signaling Impairs Cell Growth and Survival of Neurons and Progenitor Cells in the Developing Sensory Ganglia

Background: Lysophospholipids such as lysophosphatidic acid (LPA) and sphingosine-1-phosphate (S1P) are important signaling molecules that can regulate a wide range of cellular responses. We discovered that Sphingosine kinase 1 (Sphk1), a key enzyme that converts sphingosine to S1P, is expressed in neurons and progenitor cells in nascent trigeminal and dorsal root ganglia during mouse embryogenesis. Methods and Findings: Sphk1 null mouse embryos do not display overt deficits owing to compensation by Sphk2. Thus, we analyzed embryos that are deficient in both Sphk1 and Sphk2 (which essentially eliminates S1P function) in order to investigate the role(s) of Sphk1 during sensory ganglia formation. While animals lacking 1–3 alleles of Sphk1 and Sphk2 had no obvious phenotype, embryos without both genes displayed clear developmental defects. The complete absence of Sphk1 and Sphk2 resulted in trigeminal and dorsal root ganglia with fewer neurons and progenitor cells. The profound loss in cell number could be attributed to a decrease in cell proliferation as well as an increase in apoptosis. Furthermore, Sphk1/ 2 double mutants displayed an overall reduction in other sphingolipids as well as an imbalance of S1P/sphingosine and S1P/ ceramide ratio, thereby favoring cell death and reducing cell growth. Conclusions: Together, these results provide strong in vivo evidence that sphingosine kinase/S1P signaling plays a

Stellar and Gaseous Nuclear Disks Observed in Nearby (U)LIRGs

We present near-infrared integral field spectroscopy of the central kiloparsec of 17 nearby luminous and ultra-luminous infrared galaxies undergoing major mergers. These observations were taken with OSIRIS assisted by the Keck I and II Adaptive Optics systems, providing spatial resolutions of a few tens of parsecs. The resulting kinematic maps reveal gas disks in at least 16 out of 19 nuclei and stellar disks in 11 out of 11 nuclei observed in these galaxy merger systems. In our late-stages mergers, these disks are young (stellar ages $<30$ Myr) and likely formed as gas disks which became unstable to star formation during the merger. On average, these disks have effective radii of a few hundred parsecs, masses between $10^{8}$ and $10^{10} M_{Sun}$, and $v/\sigma$ between 1 and 5. These disks are similar to those created in high-resolution hydrodynamical simulations of gas-rich galaxy mergers, and favor short coalescence times for binary black holes. The few galaxies in our sample in earlier stages of mergers have disks which are larger ($r_{eff}\sim200-1800$ pc) and likely are remnants of the galactic disks that have not yet been completely disrupted by the merger.Comment: accepted for publication in Ap

Simplifying intensity-modulated radiotherapy plans with fewer beam angles for the treatment of oropharyngeal carcinoma.

The first aim of the present study was to investigate the feasibility of using fewer beam angles to improve delivery efficiency for the treatment of oropharyngeal cancer (OPC) with inverse-planned intensity-modulated radiation therapy (IP-IMRT). A secondary aim was to evaluate whether the simplified IP-IMRT plans could reduce the indirect radiation dose. The treatment plans for 5 consecutive OPC patients previously treated with a forward-planned IMRT (FP-IMRT) technique were selected as benchmarks for this study. The initial treatment goal for these patients was to deliver 70 Gy to &gt; or = 95% of the planning gross tumor volume (PTV-70) and 59.4 Gy to &gt; or = 95% of the planning clinical tumor volume (PTV-59.4) simultaneously. Each case was re-planned using IP-IMRT with multiple beam-angle arrangements, including four complex IP-IMRT plans using 7 or more beam angles, and one simple IMRT plan using 5 beam angles. The complex IP-IMRT plans and simple IP-IMRT plans were compared to each other and to the FPIMRT plans by analyzing the dose coverage of the target volumes, the plan homogeneity, the dose-volume histograms of critical structures, and the treatment delivery parameters including delivery time and the total number of monitor units (MUs). When comparing the plans, we found no significant difference between the complex IP-IMRT, simple IP-IMRT, and FP-IMRT plans for tumor target coverage (PTV-70: p = 0.56; PTV-59.4: p = 0.20). The plan homogeneity, measured by the mean percentage isodose, did not significantly differ between the IP-IMRT and FP-IMRT plans (p = 0.08), although we observed a trend toward greater inhomogeneity of dose in the simple IP-IMRT plans. All IP-IMRT plans either met or exceeded the quality of the FP-IMRT plans in terms of dose to adjacent critical structures, including the parotids, spinal cord, and brainstem. As compared with the complex IP-IMRT plans, the simple IP-IMRT plans significantly reduced the mean treatment time (maximum probability for four pairwise comparisons: p = 0.0003). In conclusion, our study demonstrates that, as compared with complex IP-IMRT, simple IP-IMRT can significantly improve treatment delivery efficiency while maintaining similar target coverage and sparing of critical structures. However, the improved efficiency does not significantly reduce the total number of MUs nor the indirect radiation dose

Shocked Gas in IRAS F17207-0014: ISM Collisions and Outflows

We combine optical and near-infrared AO-assisted integral field observations of the merging ULIRG IRAS F17207-0014 from the Wide-Field Spectrograph (WiFeS) and Keck/OSIRIS. The optical emission line ratios [N II]/H$\alpha$, [S II]/H$\alpha$, and [O I]/H$\alpha$ reveal a mixing sequence of shocks present throughout the galaxy, with the strongest contributions coming from large radii (up to 100% at $\sim$5 kpc in some directions), suggesting galactic-scale winds. The near-infrared observations, which have approximately 30 times higher spatial resolution, show that two sorts of shocks are present in the vicinity of the merging nuclei: low-level shocks distributed throughout our field-of-view evidenced by an H$_{2}$/Br$\gamma$ line ratio of $\sim$0.6-4, and strong collimated shocks with a high H$_{2}$/Br$\gamma$ line ratio of $\sim$4-8, extending south from the two nuclear disks approximately 400 pc ($\sim$0.5 arcsec). Our data suggest that the diffuse shocks are caused by the collision of the interstellar media associated with the two progenitor galaxies and the strong shocks trace the base of a collimated outflow coming from the nucleus of one of the two disks.Comment: accepted to MNRA

Following Black Hole Scaling Relations Through Gas-Rich Mergers

We present black hole mass measurements from kinematic modeling of high-spatial resolution integral field spectroscopy of the inner regions of 9 nearby (ultra-)luminous infrared galaxies in a variety of merger stages. These observations were taken with OSIRIS and laser guide star adaptive optics on the Keck I and Keck II telescopes, and reveal gas and stellar kinematics inside the spheres of influence of these supermassive black holes. We find that this sample of black holes are overmassive ($\sim10^{7-9}$ M$_{Sun}$) compared to the expected values based on black hole scaling relations, and suggest that the major epoch of black hole growth occurs in early stages of a merger, as opposed to during a final episode of quasar-mode feedback. The black hole masses presented are the dynamical masses enclosed in $\sim$25pc, and could include gas which is gravitationally bound to the black hole but has not yet lost sufficient angular momentum to be accreted. If present, this gas could in principle eventually fuel AGN feedback or be itself blown out from the system.Comment: accepted to Ap

Multiplex giant magnetoresistive biosensor microarrays identify interferon-associated autoantibodies in systemic lupus erythematosus.

High titer, class-switched autoantibodies are a hallmark of systemic lupus erythematosus (SLE). Dysregulation of the interferon (IFN) pathway is observed in individuals with active SLE, although the association of specific autoantibodies with chemokine score, a combined measurement of three IFN-regulated chemokines, is not known. To identify autoantibodies associated with chemokine score, we developed giant magnetoresistive (GMR) biosensor microarrays, which allow the parallel measurement of multiple serum antibodies to autoantigens and peptides. We used the microarrays to analyze serum samples from SLE patients and found individuals with high chemokine scores had significantly greater reactivity to 13 autoantigens than individuals with low chemokine scores. Our findings demonstrate that multiple autoantibodies, including antibodies to U1-70K and modified histone H2B tails, are associated with IFN dysregulation in SLE. Further, they show the microarrays are capable of identifying autoantibodies associated with relevant clinical manifestations of SLE, with potential for use as biomarkers in clinical practice

Diurnal Variations in Vascular Endothelial Vasodilation Are Influenced by Chronotype in Healthy Humans

Introduction: The time of day when cardiovascular events are most likely to occur is thought to be aligned with the circadian rhythm of physiological variables. Chronotype has been shown to influence the time of day when cardiovascular events happen, with early chronotypes reported to be more susceptible in the morning and late chronotypes in the evening. However, no studies have investigated the influence of chronotype on physiological variables responsible for cardiovascular regulation in healthy individuals. Methods: 312 individuals completed the Munich ChronoType Questionnaire to assess chronotype. Twenty participants were randomly selected to continue into the main study. In a repeated-measures experiment, participants were tested between 08:00 and 10:00 h and again between 18:00 and 20:00 h. Measurements of mean arterial pressure, heart rate and vascular endothelial vasodilation via flow-mediated dilatation (FMD) were obtained at each session. Results: Individual diurnal differences in mean arterial pressure and heart rate show no significant relationship with chronotype. Diurnal differences in FMD showed a significant correlation (p = 0.010), driven by a clear significant relationship in the evening and not the morning (p &lt; 0.001). Conclusion: These preliminary data indicate that chronotype influences the diurnal variation of endothelial vasodilation measured using flow-mediated dilatation. Furthermore, we show that the influence of chronotype is much stronger in the evening, highlighting an increased susceptibility for later types. These findings are consistent with the diurnal rhythm in cardiovascular events and uncover potential mechanisms of local mediators that may underpin the influence of chronotype in the onset of these events