News Article | April 26, 2017
A new type of microscope, capable of illuminating living cell structures in clear detail, could provide insight into autoimmune diseases and lead to new treatment options. Researchers from the University of Colorado Anschutz Medical Campus believe this new tool— a custom Stimulated Emission Depletion (STED) microscope—could set the stage for future treatment discoveries and visualize antibodies that cause the rare autoimmune disorder neuromyelitis optica, which can result in paralysis and blindness. The researchers used the microscope to actually see the clusters of antibodies atop astrocytes—the brain cell target of the autoimmune response in the disease. “By applying this novel approach we can see firsthand how these antibodies work,” the study's lead author, John Soltys, a current student in the Medical Scientist Training Program at CU Anschutz, said in a statement. “We are looking at the initiation of autoimmune injury in this disease.” Dr. Jeffrey Bennett, Ph.D., the senior author of the study and the associate director of Translational Research at the Center for NeuroScience at CU Anschutz, explained that the microscope allows researchers to view the early stages of various diseases as they form. “We discovered that we could see the natural clustering of antibodies on the surface of target cells,” Bennett said in a statement. “This could potentially correspond with their ability to damage the cells. “We know that once antibody binds to the surface of the astrocyte, we are witnessing the first steps in the disease process,” he added. According to Bennett, the ability to see the antibodies on the brain cells offers researchers an opportunity to develop targeted therapies that do not suppress the body’s immune system like some current treatments for the disease do. The STED microscope—which was built by CU Anschutz physicist Stephanie Meyer, Ph.D.—uses lasers to achieve a higher level of precision and clarity. Lower resolution microscopes are blurry because of the diffraction of light. However, the lasers illuminate a smaller area to acquire a higher resolution image than traditional microscopes. The STED microscope can also highlight entire living cells at extremely high resolution, unlike electron microscopes. “This would have been impossible to see with any kind of normal microscope,” professor Diego Restrepo, Ph.D., director of the Center for NeuroScience and study co-author, said in a statement. “We are inviting other scientists with research projects on campus to use the STED microscope.” The study was published in Biophysical Journal.
News Article | April 26, 2017
AURORA, Colo. (April 26, 2017) - Using a unique microscope capable of illuminating living cell structures in great detail, researchers at the University of Colorado Anschutz Medical Campus have found clues into how a destructive autoimmune disease works, setting the stage for more discoveries in the future. The scientists were trying to visualize antibodies that cause neuromyelitis optica (NMO), a rare autoimmune disorder that results in paralysis and blindness. Using a custom STED (Stimulated Emission Depletion) microscope built at CU Anschutz, they were able to actually see clusters of antibodies atop astrocytes, the brain cell target of the autoimmune response in NMO. "We discovered that we could see the natural clustering of antibodies on the surface of target cells. This could potentially correspond with their ability to damage the cells," said Professor Jeffrey Bennett, MD, PhD, senior author of the study and associate director of Translational Research at the Center for NeuroScience at CU Anschutz. "We know that once antibody binds to the surface of the astrocyte, we are witnessing the first steps in the disease process." When that domino effect begins, it's hard to stop. But Bennett said the ability to see the antibodies on the brain cells offers a chance to develop targeted therapies that do not suppress the body's immune system like current treatments for the disease do. "By applying this novel approach we can see firsthand how these antibodies work," said the study's lead author, John Soltys, a current student in the Medical Scientist Training Program at CU Anschutz. "We are looking at the initiation of autoimmune injury in this disease." The breakthrough was made possible with the STED microscope, a complex instrument that uses lasers to achieve extreme precision and clarity. It was built by physicist Stephanie Meyer, PhD, at CU Anschutz. This is the first time it has been used in a research project here. "This would have been impossible to see with any kind of normal microscope," said study co-author Professor Diego Restrepo, PhD, director of the Center for NeuroScience. "We are inviting other scientists with research projects on campus to use the STED microscope." According to Meyer, lower resolution microscopes are blurrier than the STED due to diffraction of light. But the STED's lasers illuminate a smaller area to acquire a higher resolution image. Unlike electron microscopes, STED users can see entire living cells at extremely high resolution, as they did in this study. Restrepo said there are only a handful of STEDs in the nation and just one in Colorado. The researchers said the discovery is the result of a unique partnership between clinical neurology, immunology and neuroscience coming together to solve a fundamental question of how antibodies can initiate targeted injury in an autoimmune disease. "These are the building blocks that we can use to carry our research to the next level," Bennett said. The study was published this week in Biophysical Journal.
Troutman T.D.,University of Texas Southwestern Medical Center |
Hu W.,University of Texas Southwestern Medical Center |
Fulenchek S.,University of Texas Southwestern Medical Center |
Yamazaki T.,Tokushima University |
And 4 more authors.
Proceedings of the National Academy of Sciences of the United States of America | Year: 2012
Toll like receptors (TLRs) use Toll-IL-1 receptor (TIR) domain-containing adapters, such as myeloid differentiation primary response gene 88 (MyD88) and TIR domain-containing adapter inducing IFN-β (TRIF), to induce activation of transcription factors, including NF-κB, MAP kinases, and IFN regulatory factors. TLR signaling also leads to activation of PI3K, but the molecular mechanism is not understood. Here we have discovered a unique role for B-cell adapter for PI3K (BCAP) in the TLR-signaling pathway. We find that BCAP has a functional N-terminal TIR homology domain and links TLR signaling to activation of PI3K. In addition, BCAP negatively regulates proinflammatory cytokine secretion upon TLR stimulation. In vivo, the absence of BCAP leads to exaggerated recruitment of inflammatory myeloid cells following infections and enhanced susceptibility to dextran sulfate sodium-induced colitis. Our results demonstrate that BCAP is a unique TIR domain-containing TLR signaling adapter crucial for linking TLRs to PI3K activation and regulating the inflammatory response.
Marc D.T.,NeuroScience Inc. |
Ailts J.W.,NeuroScience Inc. |
Campeau D.C.A.,NeuroScience Inc. |
Bull M.J.,NeuroScience Inc. |
Olson K.L.,NeuroScience Inc.
Neuroscience and Biobehavioral Reviews | Year: 2011
Strategies for managing the nervous system are numerous while methods of evaluating the nervous system are limited. Given the physiological importance of neurotransmitters as signaling molecules in the nervous system, the measurement of neurotransmitters has significant potential as a clinical tool. Of all the biological fluids that can be utilized, urinary neurotransmitter testing, due to its stability, sensitivity, and non-invasiveness, is the desired method to analyze nervous system function. Increasing use of this technology in a clinical setting demands a review of its feasibility, utility, and clinical value. We review the current body of literature pertaining to the mechanism of neurotransmitter transport across the blood-brain barrier as well as neurotransmitter filtration and excretion by the kidneys. In addition, this review summarizes the historical use of urinary neurotransmitter assessment to diagnose pheochromocytoma. Early research also correlated urinary assessment of neurotransmitters to various clinical symptoms and treatments of which we present research only for depression, ADHD, and inflammation because of the abundant amount of research in these areas. Finally, we review the limitations and challenges of urinary neurotransmitter testing. Taken together, evidence suggests that neurotransmitters excreted in the urine may have a place in clinical practice as a biomarker of nervous system function to effectively assess disturbances and monitor treatment efficacy. © 2010 Elsevier Ltd.
Ma X.,Genentech |
Lin W.Y.,Genentech |
Chen Y.,Genentech |
Stawicki S.,Genentech |
And 6 more authors.
Structure | Year: 2012
Lacking any discernible sequence similarity, interleukin-34 (IL-34) and colony stimulating factor 1 (CSF-1) signal through a common receptor CSF-1R on cells of mononuclear phagocyte lineage. Here, the crystal structure of dimeric IL-34 reveals a helical cytokine fold homologous to CSF-1, and we further show that the complex architecture of IL-34 bound to the N-terminal immunoglobulin domains of CSF-1R is similar to the CSF-1/CSF-1R assembly. However, unique conformational adaptations in the receptor domain geometry and intermolecular interface explain the cross-reactivity of CSF-1R for two such distantly related ligands. The docking adaptations of the IL-34 and CSF-1 quaternary complexes, when compared to the stem cell factor assembly, draw a common evolutionary theme for transmembrane signaling. In addition, the structure of IL-34 engaged by a Fab fragment reveals the mechanism of a neutralizing antibody that can help deconvolute IL-34 from CSF-1 biology, with implications for therapeutic intervention in diseases with myeloid pathogenic mechanisms. © 2012 Elsevier Ltd.
Asherson P.,King's College London |
Bushe C.,Eli Lilly and Company |
Saylor K.,Neuroscience Inc. |
Tanaka Y.,Eli Lilly and Company |
And 2 more authors.
Journal of Psychopharmacology | Year: 2014
Persistence of attention deficit hyperactivity disorder (ADHD) into adulthood can be disabling or lead to substantial impairment. Several clinical trials of atomoxetine (ATX) in adults with ADHD have been reported following the National Institute for Health and Clinical Excellence (NICE) guidelines issued in 2008. We performed an integrated analysis of all Eli Lilly-sponsored, randomized, double-blind, placebo-controlled studies of ATX in adults with ADHD completed as of May 2012. Individual patient data were pooled from six short-term (10-16 week) studies (1961 patients) and three longer-term (six-month) studies (1413 patients). In the short-term analysis, ATX patients achieved a significantly greater mean reduction in ADHD symptoms than placebo patients (-12.2 vs -8.1; Conners' Adult ADHD Rating Scale-Investigator-Rated: Screening Version (CAARS-Inv: SV); p<0.001). In the longer-term analysis, respective improvements after six months were -13.2 vs -9.7 (p<0.001). Response rates at study endpoints for ATX vs placebo, based on CAARS-Inv: SV improvement ≥30% and Clinical Global Impressions of ADHD-Severity (CGI-ADHD-S) ≥3 were 34.8% vs 22.3% in the short-term and 43.4% vs 28.0% after six months, and CAARS-Inv: SV improvements ≥40% were 41.3% vs 25.3% in the short-term and 44.0% vs 31.4% after six months (all p<0.001). Overall, ATX had a clinically significant effect in adults with ADHD, with reductions in core symptoms and clinically meaningful responder rates. © The Author(s) 2014.
Nichkova M.,Pharmasan Labs Inc. |
Wynveen P.M.,Pharmasan Labs Inc. |
Marc D.T.,NeuroScience Inc. |
Huisman H.,Pharmasan Labs Inc. |
Kellermann G.H.,NeuroScience Inc.
Journal of Neurochemistry | Year: 2013
Dopamine is a catecholamine that serves as a neurotransmitter in the central and peripheral nervous system. Non-invasive, reliable, and high-throughput techniques for its quantification are needed to assess dysfunctions of the dopaminergic system and monitor therapies. We developed and validated a competitive ELISA for direct determination of dopamine in urine samples. The method provides high specificity, good accuracy, and precision (average inter-assay variation < 12%). The analysis is not affected by general urinary components and structurally related drugs and metabolites. The correlation between ELISA and LC-MS/MS analyses was very good (r = 0.986, n = 28). The reference range was 64-261 μg/g Cr (n = 64). Week-to-week biological variations of second morning urinary dopamine under free-living conditions were 23.9% for within- and 35.5% for between-subject variation (n = 10). The assay is applied in monitoring Parkinson's disease patients under different treatments. Urinary dopamine levels significantly increase in a dose-dependent manner for Parkinson's disease patients under l-DOPA treatment. The present ELISA provides a cost-effective alternative to chromatographic methods to monitor patients receiving dopamine restoring treatment to ensure appropriate dosing and clinical efficacy. The method can be used in pathological research for the assessment of possible peripheral biological markers for disorders related to the dopaminergic system. Our competitive ELISA for direct dopamine quantification in urine samples is a viable cost-effective alternative to chromatographic analysis. The method is robust, sensitive, and very specific and it does not require sample pre-treatment. It can be used in monitoring dopamine-modulating therapies. Urinary dopamine levels significantly increase in a dose-dependent manner for Parkinson's disease patients under l-DOPA treatment. © 2013 International Society for Neurochemistry.
Kheirandish-Gozal L.,University of Chicago |
McManus C.J.T.,NeuroScience Inc. |
Kellermann G.H.,NeuroScience Inc. |
Samiei A.,University of Chicago |
Gozal D.,University of Chicago
Chest | Year: 2013
Background: Pediatric obstructive sleep apnea (OSA) is associated with cognitive dysfunction, suggesting altered neurotransmitter function. We explored overnight changes in neurotransmitters in the urine of children with and without OSA. Methods: Urine samples were collected from children with OSA and from control subjects before and after sleep studies. A neurocognitive battery assessing general cognitive ability (GCA) was administered to a subset of children with OSA. Samples were subjected to multiple enzyme-linked immunosorbent assays for 12 neurotransmitters, and adjusted for creatinine concentrations. Results: The study comprised 50 children with OSA and 20 control subjects. Of the children with OSA, 20 had normal GCA score (mean ± SD) (101.2 ± 14.5) and 16 had a reduced GCA score (87.3 ± 13.9; P<.001). Overnight increases in epinephrine, norepinephrine, and γ-aminobutyric acid (GABA) levels emerged in children with OSA; taurine levels decreased. Using combinatorial approaches and cutoff values for overnight changes of these four neurotransmitters enabled prediction of OSA (area under the curve [AUC]: 0.923; P<.0001). Furthermore, GABA and taurine alterations, as well as overnight reductions in phenylethylamine, were more prominent in children with OSA and low GCA than in children with OSA and normal GCA (P<.001), and they reliably discriminated GCA status (AUC: 0.977; P<.0001). Conclusions: Pediatric OSA is associated with overnight increases in urinary concentrations of catecholamines indicative of heightened sympathetic outflow. Increases in GABA levels and decreases in taurine levels could underlie mechanisms of neuronal excitotoxicity and dysfunction. Combinatorial approaches using defined cutoffs in overnight changes in concentrations of selected neurotransmitters in urine may not only predict OSA but also the presence of cognitive deficits. Larger cohort studies appear warranted to confirm these findings. © 2013 American College of Chest Physicians.
Troutman T.D.,University of Texas Southwestern Medical Center |
Bazan J.F.,NeuroScience Inc. |
Pasare C.,University of Texas Southwestern Medical Center
Cell Cycle | Year: 2012
TLRs are a family of pattern recognition receptors that recognize conserved molecular structures/products from a wide variety of microbes.1,2Following recognition of ligands, TLRs recruit signaling adapters to initiate a pro-inflammatory signaling cascade culminating in the activation of several transcription factor families. Additionally, TLR signals lead to activation of PI3K, affecting many aspects of the cellular response, including cell survival, proliferation and regulation of the pro-inflammatory response.3-10 The recent discovery of BCAP as a TLR signaling adaptor, crucial for linking TLRs to PI3K activation, allows new questions of the importance of PI3K activation downstream of TLRs. Here, we summarize the current understanding of signaling pathways activated by TLRs and provide our perspective on TLR mediated activation of PI3K and its impact on regulating cellular processes. © 2012 Landes Bioscience.
Neuroscience Inc. | Date: 2010-10-28
This document provides methods and materials related to managing weight, supporting appetite control, and controlling cravings associated with smoking reduction or cessation regimens and/or nicotine reduction or cessation regimens. For example, compositions comprising an agent to support acetylcholine and an agent to support one or more biogenic amines, and methods for using such compositions for craving and appetite control are provided. Methods and materials to reduce cravings associated with the reduction or cessation of the use of chemical substances (e.g., drugs of abuse, including opioids, cocaine, methamphetamine, cannabis, alcohol), and to reduce cravings associated with addictive and/or compulsive behaviors (e.g., gambling, sex, and repetitive behaviors) are also provided.