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March 2018; 5 (2) ArticleOpen Access

Defining standard enzymatic dissociation methods for individual brains and spinal cords in EAE

Rehana Z. Hussain, William A. Miller-Little, Richard Doelger, Gary R. Cutter, Nicolas Loof, Petra D. Cravens, Olaf Stüve
First published January 18, 2018, DOI: https://doi.org/10.1212/NXI.0000000000000437
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Article Information

vol. 5 no. 2 e437
DOI: 
https://doi.org/10.1212/NXI.0000000000000437
Published By: 
Wolters Kluwer Health, Inc. on behalf of the American Academy of Neurology
Online ISSN: 
2332-7812
History: 
  • Received August 13, 2017
  • Accepted in final form November 9, 2017
  • First Published January 18, 2018.
Copyright & Usage: 
Copyright © 2018 The Author(s). Published by Wolters Kluwer Health, Inc. on behalf of the American Academy of Neurology. This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND), which permits downloading and sharing the work provided it is properly cited. The work cannot be changed in any way or used commercially without permission from the journal.

Author Disclosures

    1. Rehana Z. Hussain, MSc*,
    2. William A. Miller-Little, BA*,
    3. Richard Doelger, MSc,
    4. Gary R. Cutter, PhD,
    5. Nicolas Loof, MSc,
    6. Petra D. Cravens, PhD and
    7. Olaf Stüve, MD, PhD
  1. Rehana Z. Hussain, MSc*,
    2. Scientific Advisory Boards:

    1. NONE

    Gifts:

    1. NONE

    Funding for Travel or Speaker Honoraria:

    1. NONE

    Editorial Boards:

    1. NONE

    Patents:

    1. NONE

    Publishing Royalties:

    1. NONE

    Employment, Commercial Entity:

    1. NONE

    Consultancies:

    1. NONE

    Speakers' Bureaus:

    1. NONE

    Other Activities:

    1. NONE

    Clinical Procedures or Imaging Studies:

    1. NONE

    Research Support, Commercial Entities:

    1. NONE

    Research Support, Government Entities:

    1. NONE

    Research Support, Academic Entities:

    1. NONE

    Research Support, Foundations and Societies:

    1. NONE

    Stock/Stock Options/Board of Directors Compensation:

    1. NONE

    License Fee Payments, Technology or Inventions:

    1. NONE

    Royalty Payments, Technology or Inventions:

    1. NONE

    Stock/Stock Options, Research Sponsor:

    1. NONE

    Stock/Stock Options, Medical Equipment & Materials:

    1. NONE

    Legal Proceedings:

    1. NONE

  2. William A. Miller-Little, BA*,
    3. Scientific Advisory Boards:

    1. NONE

    Gifts:

    1. NONE

    Funding for Travel or Speaker Honoraria:

    1. NONE

    Editorial Boards:

    1. NONE

    Patents:

    1. NONE

    Publishing Royalties:

    1. NONE

    Employment, Commercial Entity:

    1. NONE

    Consultancies:

    1. NONE

    Speakers' Bureaus:

    1. NONE

    Other Activities:

    1. NONE

    Clinical Procedures or Imaging Studies:

    1. NONE

    Research Support, Commercial Entities:

    1. NONE

    Research Support, Government Entities:

    1. NONE

    Research Support, Academic Entities:

    1. NONE

    Research Support, Foundations and Societies:

    1. NONE

    Stock/Stock Options/Board of Directors Compensation:

    1. NONE

    License Fee Payments, Technology or Inventions:

    1. NONE

    Royalty Payments, Technology or Inventions:

    1. NONE

    Stock/Stock Options, Research Sponsor:

    1. NONE

    Stock/Stock Options, Medical Equipment & Materials:

    1. NONE

    Legal Proceedings:

    1. NONE

  3. Richard Doelger, MSc,
    4. Scientific Advisory Boards:

    1. NONE

    Gifts:

    1. NONE

    Funding for Travel or Speaker Honoraria:

    1. NONE

    Editorial Boards:

    1. NONE

    Patents:

    1. NONE

    Publishing Royalties:

    1. NONE

    Employment, Commercial Entity:

    1. NONE

    Consultancies:

    1. NONE

    Speakers' Bureaus:

    1. NONE

    Other Activities:

    1. NONE

    Clinical Procedures or Imaging Studies:

    1. NONE

    Research Support, Commercial Entities:

    1. NONE

    Research Support, Government Entities:

    1. NONE

    Research Support, Academic Entities:

    1. NONE

    Research Support, Foundations and Societies:

    1. NONE

    Stock/Stock Options/Board of Directors Compensation:

    1. NONE

    License Fee Payments, Technology or Inventions:

    1. NONE

    Royalty Payments, Technology or Inventions:

    1. NONE

    Stock/Stock Options, Research Sponsor:

    1. NONE

    Stock/Stock Options, Medical Equipment & Materials:

    1. NONE

    Legal Proceedings:

    1. NONE

  4. Gary R. Cutter, PhD,
    5. Scientific Advisory Boards:

    1. Data and Safety Monitoring Boards: AMO Pharmaceuticals, Apotek, Horizon Pharmaceuticals, Modigenetech/Prolor, Merck, Merck/Pfizer, Opko Biologics, Neurim, Sanofi-Aventis, Reata Pharmaceuticals, Receptos/Celgene, Teva pharmaceuticals, NHLBI (Protocol Review Committee), NICHD (OPRU oversight committee).

    Gifts:

    1. NONE

    Funding for travel or speaker honoraria:

    1. Data and Safety Monitoring Boards: AMO Pharmaceuticals, Apotek, Horizon Pharmaceuticals, Modigenetech/Prolor, Merck, Merck/Pfizer, Opko Biologics, Neurim, Sanofi-Aventis, Reata Pharmaceuticals, Receptos/Celgene, Teva pharmaceuticals, NHLBI (Protocol Review Committee), NICHD (OPRU oversight committee). Consulting or Advisory Boards: Atara Biotherapeutics, Argenix, Bioeq GmBH, Consortium of MS Centers (grant), Genzyme, Genentech, Innate Therapeutics, Klein-Buendel Incorporated, Medimmune, Medday, Novartis, Opexa Therapeutics, Roche, Savara Inc., Somahlution, Teva pharmaceuticals, Transparency Life Sciences, TG Therapeutics

    Editorial Boards:

    1. Am journal of the Society of Nephrology. Statistical Reviewer

    Patents:

    1. NONE

    Publishing Royalties:

    1. NONE

    Employment, Commercial Entity:

    1. NONE

    Consultancies:

    1. Consulting or Advisory Boards: Atara Biotherapeutics, Argenix, Bioeq GmBH, Consortium of MS Centers (grant), Genzyme, Genentech, Innate Therapeutics, Klein-Buendel Incorporated, Medimmune, Medday, Novartis, Opexa Therapeutics, Roche, Savara Inc., Somahlution, Teva pharmaceuticals, Transparency Life Sciences, TG Therapeutics

    Speakers' Bureaus:

    1. NONE

    Other activities:

    1. President of Pythagoras Inc. a consulting company...gc

    Clinical Procedures or Imaging Studies:

    1. NONE

    Research Support, Commercial Entities:

    1. NONE

    Research Support, Government Entities:

    1. CUTTER, GARY R. ACTIVE No Number (Cutter) 01/01/2014 ? 12/31/2016 0.01 calendar Myasthenia Gravis Foundation of America, Inc. $NCE UC FFS - MGFA Survey and Website Development, Maintenance Support Related to the MG Patient Registry Role: Principal Investigator U19-AI113212 (Cutter) 06/15/2014 ? 05/31/2019 1.2 calendar NIH/NIAID $183,087 current year direct The UAB STI CRC ? DiSCIS Biostatistics & Bioinformatics Core This application to become a Sexually Transmitted Infections Cooperative Research Center from the University of Alabama at Birmingham is comprised of a group of innovative, multidisciplinary translational research projects with the goal of addressing the microbial origins and pathogenesis of the most prevalent genital discharge syndromes in men (non-gonococcal urethritis) and women (bacterial vaginosis), as well as the interaction of these syndromes with the most prevalent sexually transmitted bacteria, Chlamydia trachomatis. Our CRC research projects will evaluate patients from a unique cohort of sexual partners, and study findings should lead to improved or new strategies for the prevention, diagnosis, and/or management of sexually transmitted infections. Role: Project Director W81XWH-14-2-0167 (Taub) 09/30/2014 ? 09/29/2017 .30 calendar DOD-Army Medical Research Acquisition Activity $420,471 current year direct Harnessing Neuroplasticity to Promote Rehabilitation: CI Therapy for TBI The purpose of the proposed research is to determine the value of Constraint-Induced Movement therapy (CIMT) for improving motor ability in veterans/warfighters with traumatic brain injury (TBI) and test whether any therapeutic effects observed are correlated with neuroplastic white matter or grey matter changes. Role: Co-Investigator U01-HL 119242 (Cutter) 09/01/2014 ? 05/31/2020 .3 calendar NIH/NHLBI $426,203 current year direct Chronic Hypertension and Pregnancy (CHAP): Data Coordinating Center We propose a large pragmatic multi-center randomized trial of pregnant women with mild chronic hypertension to evaluate the benefits and harms of antihypertensive therapy to a goal <140/90 mmHg (as recommended for the general population in the US) compared with ACOG's current policy of expectant management of mild chronic hypertension in pregnancy. The trial will be conducted in 12 experienced research-oriented Ob/Gyn departments (including 25 clinical sites) in the United States. The monitoring plan will include a pre-specified option to increase the planned sample size of 4700 women after interim evaluation by an independent Data Safety and Monitoring Board. Role: Principal Investigator No number assigned (Cutter) 01/01/2004 ? 12/31/2016 0.01 calendar Consortium of MS Centers (CMSC) $ 600,000 North American Research Consortium on Multiple Sclerosis (NARCOMS) The goals of this project are to facilitate a confidential way for patients to supply valuable information to researchers about their course of disease that may lead to more effective treatments and care for people living with MS, while reducing the time and cost of conducting studies; provide a worldwide research resource for people living with multiple sclerosis so they can provide information about themselves and their course of disease; and develop new collaborations between researchers, patients, and healthcare providers. UAB provides programming and analytic support for the NARCOMS registry. Role: Principal Investigator W81XWH-12-1-0155 (Korf) 05/15/2012 ? 05/14/2017 1.1 calendar U.S. Department of Defense $1,621,899 current year direct NEUTOFIBOMATOSIS CLINICAL CONSORTIUM AWARD This is a cooperative study group that is focusing on multiple trials in NF. The role of the operations center is both as the data center and the overall coordinating of the study group. Role: Co-Investigator P30DK079337 (Agarwal) 09/20/13 ? 07/31/18 .30 calendar NIH/NIDDK $976,357 current year direct UAB/UCSD O?Brien Core Center for Acute Kidney Injury Research In summary, these cores and the outstanding cohort of investigators assembled for this center will provide unique expertise that is critical for innovative and productive research in AKI. With its Extended Research Base that includes both clinical and basic investigators, this O'Brien center will accelerate the translation of new investigative insights towards novel therapies for patients with AKI. Role: Co-Investigator 5U01NS082320 (Cutter) 09/01/13 ? 08/31/17 .3 calendar NIH/ Children?s Hospital (Boston) $60,314 current year direct Early Biomarkers of Autism Spectrum Disorders in Infants with Tuberous Sclerosis Studying infants with Tuberous Sclerosis Complex (TSC), a genetic disorder with a high prevalence of ASD that can be diagnosed before or at the time of birth, allows us to develop better tools for early detection of autism. We are investigating whether longitudinal assessment of brain connectivity using MRI and EEG in children with TSC can identify an early biomarker of subsequent ASD in genetic disease. This will result in better understanding of brain connectivity and its relationship to ASD in TSC and will pave the way for new interventions and this and related causes of autism. Role: Principal Investigator of Subcontract Site No Number (Standaert) 04/01/14 ? 09/30/17 0.6 calendar Alabama Department of Commerce $666,667 current year direct UAB Cannabidiol Program The goal of this project is to establish a program for cannabidiol treatment for seizures Role: Co-Investigator P2CHD086851 (Bamman) 09/17/2015 ? 06/30/2020 2.1 calendar NIH/NICHHD $778,111 current year direct National Resource Center for High-Impact Clinical Trials in Medical Rehabilitation There are major knowledge gaps in medical rehabilitation that demand clinical trials of the highest quality. With particular emphasis on chronic conditions such as stroke, brain or spinal cord injury, orthopedic and joint conditions, and developmental and degenerative disorders leading to disability, the overarching purpose of the proposed P2C Medical Rehabilitation Research Resource, based at The University of Alabama at Birmingham (UAB), is to catalyze high-impact clinical trials that will define optimal rehabilitation strategies. Role: Co-Investigator 1U01EY025858 (Visscher) 05/01/2016 ? 04/30/2020 .6 calendar NIH/National Eye Institute $287,298 current year direct Changes in Visual Cortical Connectivity Following Central Visual Field Loss When people lose central vision, they must use peripheral vision to do daily tasks like reading and recognizing faces. This proposal will examine how changes in the connectivity and structure of early visual cortex relate to this huge shift in experience. This is relevant to public health because it develops fundamental knowledge about how the visual system adapts after vision loss and this knowledge may be used to advance future therapies. Role: Co-Investigator 1U01NS092595 (Bebin) 08/04/2016 ? 08/03/2021 1.2 calendar NIH/NINDS $1,243,905 current year direct PREVENTING EPILEPSY USING VIGABATRIN IN INFANTS WITH TUBEROUS SCLEROSIS COMPLEX The primary objective of the study is the developmental impact of early versus delayed treatment with vigabatrin at 24 months of age based on the cognitive assessment score of the Bayley-III. The secondary study objectives will focus on the effectiveness of early versus delayed treatment with vigabatrin in clinical seizure prevention and the prevalence of drug resistant epilepsy by the age of 24 months. Also, the impact of early versus delayed vigabatrin treatment on receptive communication, expressive communication, fine and gross motor skills, and risk of autism spectrum disorders (ASD). Role: Co-Investigator No Number (Corboy) 05/16/2016 ? 11/17/2020 0.3 calendar Patient-Centered Outcomes Research Institute (PCORI) $171,744 current year direct Discontinuation of Disease Modifying Therapies (DMTs) in Multiple Sclerosis (MS) A significant information gap exists about a condition that is both significantly debilitating to patients, and costly to those afflicted with MS and to society. There may be both risks and benefits to the patient when continuing or discontinuing MS DMTs, and patients may view risks and benefits quite differently from their physician. It appears the MS population that has the lowest risk, and perhaps is most likely to benefit, from potential discontinuation of MS DMTs, is relatively older patients with no recent evidence of new inflammatory disease activity. We propose to study deliberate discontinuation in this subpopulation of MS patients, to determine if DMT discontinuation is safe and results in no worse physician-derived and patient-reported outcomes. Role: Co-Investigator UH3NS100553 (Walker) 09/30/16 ? 06/30/21 1.2 calendar NIH $980,858 Noninvasive Biomarkers to Advance Emerging DBS Electrode Technologies in Parkinson?s Disease The goal of this research is to use minimally invasive, patient-specific cortical physiology elicited by DBS to guide the use of emerging segmented (?directional?) DBS electrode technology in patients with Parkinson?s disease. Role: Co-Investigator

    Research Support, Academic Entities:

    1. NONE

    Research Support, Foundations and Societies:

    1. NONE

    Stock/Stock Options/Board of Directors Compensation:

    1. Consortium of MS Centers - President - only expenses covered

    License Fee Payments, Technology or Inventions:

    1. NONE

    Royalty Payments, Technology or Inventions:

    1. NONE

    Stock/Stock Options, Research Sponsor:

    1. NONE

    Stock/Stock Options, Medical Equipment & Materials:

    1. NONE

    Legal Proceedings:

    1. NONE

  5. Nicolas Loof, MSc,
    6. Scientific Advisory Boards:

    1. NONE

    Gifts:

    1. NONE

    Funding for Travel or Speaker Honoraria:

    1. NONE

    Editorial Boards:

    1. NONE

    Patents:

    1. NONE

    Publishing Royalties:

    1. NONE

    Employment, Commercial Entity:

    1. NONE

    Consultancies:

    1. NONE

    Speakers' Bureaus:

    1. NONE

    Other Activities:

    1. NONE

    Clinical Procedures or Imaging Studies:

    1. NONE

    Research Support, Commercial Entities:

    1. NONE

    Research Support, Government Entities:

    1. NONE

    Research Support, Academic Entities:

    1. NONE

    Research Support, Foundations and Societies:

    1. NONE

    Stock/Stock Options/Board of Directors Compensation:

    1. NONE

    License Fee Payments, Technology or Inventions:

    1. NONE

    Royalty Payments, Technology or Inventions:

    1. NONE

    Stock/Stock Options, Research Sponsor:

    1. NONE

    Stock/Stock Options, Medical Equipment & Materials:

    1. NONE

    Legal Proceedings:

    1. NONE

  6. Petra D. Cravens, PhD and
    7. Scientific Advisory Boards:

    1. NONE

    Gifts:

    1. NONE

    Funding for Travel or Speaker Honoraria:

    1. NONE

    Editorial Boards:

    1. NONE

    Patents:

    1. NONE

    Publishing Royalties:

    1. NONE

    Employment, Commercial Entity:

    1. NONE

    Consultancies:

    1. NONE

    Speakers' Bureaus:

    1. NONE

    Other Activities:

    1. NONE

    Clinical Procedures or Imaging Studies:

    1. NONE

    Research Support, Commercial Entities:

    1. NONE

    Research Support, Government Entities:

    1. NONE

    Research Support, Academic Entities:

    1. NONE

    Research Support, Foundations and Societies:

    1. NONE

    Stock/Stock Options/Board of Directors Compensation:

    1. NONE

    License Fee Payments, Technology or Inventions:

    1. NONE

    Royalty Payments, Technology or Inventions:

    1. NONE

    Stock/Stock Options, Research Sponsor:

    1. NONE

    Stock/Stock Options, Medical Equipment & Materials:

    1. NONE

    Legal Proceedings:

    1. NONE

  7. Olaf Stüve, MD, PhD
    8. Scientific Advisory Boards:

    1. Pfizer, TG Therapeutics

    Gifts:

    1. NONE

    Funding for Travel or Speaker Honoraria:

    1. NONE

    Editorial Boards:

    1. Therapeutic Advances in Neurological Disorders Multiple Sclerosis Journal

    Patents:

    1. NONE

    Publishing Royalties:

    1. NONE

    Employment, Commercial Entity:

    1. NONE

    Consultancies:

    1. NONE

    Speakers' Bureaus:

    1. NONE

    Other Activities:

    1. NONE

    Clinical Procedures or Imaging Studies:

    1. NONE

    Research Support, Commercial Entities:

    1. Teva Pharmaceuticals Opexa Therapeutics

    Research Support, Government Entities:

    1. Merit Review grant (federal award document number (FAIN) I01BX001674) from the United States (U.S.) Department of Veterans Affairs, Biomedical Laboratory Research and Development

    Research Support, Academic Entities:

    1. NONE

    Research Support, Foundations and Societies:

    1. NONE

    Stock/Stock Options/Board of Directors Compensation:

    1. NONE

    License Fee Payments, Technology or Inventions:

    1. NONE

    Royalty Payments, Technology or Inventions:

    1. NONE

    Stock/Stock Options, Research Sponsor:

    1. NONE

    Stock/Stock Options, Medical Equipment & Materials:

    1. NONE

    Legal Proceedings:

    1. NONE

  1. From the Department of Neurology and Neurotherapeutics (R.Z.H., W.A.M.-L., R.D., P.D.C., O.S.), University of Texas Southwestern Medical Center, Dallas; Department of Biostatistics (G.C.), University of Alabama at Birmingham; The Moody Foundation Flow Cytometry Facility (N.L.), Children's Research Institute, University of Texas Southwestern Medical Center, Dallas; Neurology Section (O.S.), VA North Texas Health Care System, Medical Service, Dallas, TX; and Department of Neurology (O.S.), Klinikum rechts der Isar, Technische Universität München, Germany.
  1. Correspondence
    Dr. Stüve olaf.stuve{at}utsouthwestern.edu
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