BIO 1000 Assessment Neuromuscular Lab

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BIO 1000 Assessment Neuromuscular Lab

BIO 1000 Assessment Neuromuscular Lab

For this assessment, complete the Neuromuscular Lab linked
in the Resources, under the Capella Multimedia heading. The Neuromuscular Lab
focuses on three patients who are getting their yearly physical, including an
eye exam and reflex test. It will have you go through the process of conducting
the eye exams from an optometrist’s point of view.

At the end of the lab is an assessment. To complete the
assessment, fill out the items requested within the lab, and then download your
results to your computer. Then submit this lab results file as your assessment.
Since all of your work for the assessment is completed within the lab, you are
advised to examine the assessment scoring guide prior to completing the lab and
downloading your lab results.

NEUROMUSCULAR CLINICAL LABORATORY

Contact Information
Address
MSC 8111-0009-04
660 South Euclid Avenue
St. Louis, MO 63110
Phone: 314-362-2406
Fax: 314-362-3413
e-mailnmlab@wustl.edu


Instructions for sending
Antibody testing
, or
Biopsies: Muscle or Nerve
Phone: 314-362-2406


Laboratory certifications
CAP Lab # 19233-16
Medicare Provider # 26-8235
CLIA Certificate # 26D0652044
New York State 3499
California COS 00800679
Florida 800028650
Pennsylvania 37448

Testing at Washington University
Laboratory: Specific tests
Requisition forms & Information
Serum (Antibody & Antigen)
Sending samples: Instructions
Biopsies: Muscle, Nerve & Skin
Muscle instructions
FreezingOther
Nerve instructions
GeneralSurgery;
FreezingOther technical
Skin instructions

Antibodies
General principles
Classification
Binding to oligosaccharides
Glycoproteins & Glycolipids
Specific types
Testing methods
Survey


Charcot's clinic
Charcot’s clinic


 


Testing at Washington University Neuromuscular Laboratory


Antibodies


Muscle biopsies

  • Muscle biopsies can be sent fresh or frozenInstructions
  • Tests performed
    • Histochemical stains include:
      H&E; GT; NADH; ATPase, pH9.4, 4.6 & 4.3; Congo red; VvG;
      Alkaline phosphatase; Acid phosphatase; Esterase; Cytochrome oxidase; SDH
      AMPDA; Phosphorylase; Phosphofructokinase; Sudan black; PAS
    • Dystrophy-related immunostains (Immunohistochemistry and/or Western blot) include:
      Dystrophin (Dys 1, 2, 3); Sarcoglycans (α, β, γ, δ); Dysferlin; α-Dystroglycan; Lamin A/C
      Merosin; Caveolin-3; Calpain-3; Desmin; Emerin; α-actinin; LAMP-2; MYH2; Actin (Phalloidin)
      Laminin-α2; laminin-B; Collagen VI & IV; Phosphorylated epitopes (Neurofilament & other; SMI-31 antibody)
    • Immune myopathy panel
      MHC Class I; Capillary morphology (Ulex); C5b-9 (MAC);
      CD4; CD20; CD8; HAM56; CD163; Decorin; SMI-31; LC3; CA9; Alcian blue
    • Aggregate stains: Desmin; VCP; Phalloidin; Caveolin-3; Ubiquitinated proteins (FK2); SMI-31; TDP-43
    • Mitochondria: Oxidative-phosphorylation enzyme complex activities + Citrate Synthase + Coenzyme Q10 levels
    • Glycogen pathway: Glycogen; Acid Maltase; Glycogen degradation (PFK, Phosphorylase, PGK, PGM1, PGAM1, LDH, PBK); PFK stain
    • Plastic sections (Toluidine blue stained): When necessary
      • Ultrastructure: When necessary
  • Requisition form: Printable PDF
  • Information about muscle biopsies
  • Certification: Alan Pestronk MD

Nerve biopsies

  • Nerve biopsy specimens: Preparation
    • Size
      • Full thickness
      • Length: > 3 cm
    • Fresh: Transported in moist gauze on wood stick
    • Processed: 3 pieces of equal size
      • Frozen
      • Formalin fixed: 10% Buffered Formalin
      • Glutaraldehyde fixed
        • Modified Karnovsky’s Fixative: (3% Glutaraldehyde, 1% Paraformaldehyde in 0.1M sodium cacodylate buffer)
  • Nerve biopsies: Evaluation
    • Frozen sections
      • Stains: H&E; GT; Alkaline phosphatase; Acid phosphatase; Congo red; VvG; Neurofilament (Axons); NCAM (Schwann cells)
      • Immune features
    • Plastic sections: Toluidine blue stained
      • Ultrastructure: When necessary
    • Paraffin sections: H&E
  • Requisition form: Printable PDF
  • Nerve biopsies: Information
  • Certification: Alan Pestronk MD

Neuromuscular Pathology Training Program

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LAB TESTING IN NEUROMUSCULAR DISORDERS
Antibodies Biopsies WU Testing
IgM
Decorin (BJ)
Disaccharides
NS6S
TS-HDS
Glycolipids
GalC
GalNAc-GD1a
GALOP
GD1a
GD1b
GM1
GM1b
GM2
NP9
SGPG
Sulfatide
Heparan sulfate
MAG
Neurofascin-155
Neurofilaments
Tubulin
IgG
Acetylcholine receptors
Muscle
Neuronal (α3)
Other
NMJ
LRP4
MuSK
Channels
Calcium
Potassium complex
Lgi1
Caspr2
Paranodal
Caspr
Contactin-1
Neurofascins
140155
Cortactin
CRMP-5
DPPX
FGFR-3
FHL1
GAD
Glutamate, Glycine &
GABA Receptor

Glycolipids
GalC
GM1
GD1a
GalNAc-GD1a
Disialosyl
GD1b
GQ1b
GT1a
Lysoganglioside-GM1
Sulfatide
Mitochondria
Myopathy
HMGCR
Jo-1
MDA5
Muscle striations
NT5C1A
Ro
Myositis
MPP1
Neuronal glia
SOX1
Neuronal nuclei
Hu
Ri
Plexin D1
Purkinje cells
PCA-2
Tr
Yo
Antibodies: General
Testing methods
CNS-related
Connective tissue disorders
Myositis-associated
Myositis-overlap
Other antibodies
Vasculitis
Diabetes
M-proteins
Myopathies
Myositis
Neuropathies
NMJ disorders
Myasthenia Gravis
LEMS
Oligosaccharide antigens
Paraneoplastic
Stiffman
Transglutaminase
Types
Polyspecific IgM & IgG
IgG4


Other Tests
Neurofilament Heavy Chain (CSF)

Muscle

Nerve

Skin

Antibodies
Requisition
Instructions

Biopsies
Requisition

Laboratory testing at Washington University, St. Louis

 


Neuromuscular Autoantibodies: Classification & Features

Antibody Antigen Disease
Class Location Type Pathogenic
properties
Features Antigen
Examples
Neuromuscular
Disease Examples
Associations,
Systemic
Common
treatments
IgG Serum Polyclonal Often Protein
Cell surface
membrane
Receptors: AChRs
Muscle; Neuron
Ca++ ion channels
K+ channel complex
Myasthenia gravis
Autonomic PN
LEMS
Isaac’s
Neoplasms
Thymoma
Adeno Ca
Small cell lung
Corticosteroids
T-cell suppressants
IVIg
Plasma Exchange
IgG Serum Polyclonal Possible Carbohydrate
Glycolipid
Gangliosides
GM1
GQ1b
GD1b
AMAN
Miller-Fisher
Sensory, Acute
Infection,
Prodromal
Disease often
monophasic
IgG Serum Polyclonal Few Protein
Intracellular
tRNA synthetases
SRP
ANCA
FGFR3
IMPP
Polymyopathy, immune
Vasculitis
Neuropathy, Sensory
Organ system
disease, or
Neoplasm, or
None
Corticosteroids
T-cell suppressants
Rituximab
IgG Serum +
CSF
Polyclonal No Protein
Intracellular
Hu (Neuron nuclei)

CRMP5

Neuronopathy
Sensory; Autonomic
Polyneuropathy
Neoplasms
Small cell lung
Gynecologic
Screen for & Treat
associated Neoplasm
IgG4 Serum Polyclonal Possible Protein
Cell surface, or
Extracellular
MuSK
Neurofascins
140
155
Contactin-1
Lgi1
Caspr2
MG-MuSK
PN
Sensory ataxia, subacute
Demyel +: TremorCNS
Demyel PN: IVIg resistant
Encephalopathy ± PN
NeuromyotoniaAutonomic
None Rituximab (q 1-6 mo)
Corticosteroids
IgM Serum Monoclonal
or
Polyclonal
Possible Carbohydrate
Glycolipid or
Glycoproteins


Protein

GM1
MAG
TS-HDS
GD1b


Neurofascin-155

Motor PN
Demyelinating PN
Sensory PN
Sensory PN, Ataxic


SM-PN, Demyel + Tremor

Neoplasms
M-protein


No

Rituximab (IgM ↓)
Intravenous Ig
(Conduction block)
Cyclophosphamide


Intravenous Ig

IgM, IgG,
or IgA
Serum Monoclonal Some Not identified Polyneuropathy POEMS
Cryoglobulins
Waldenström’s
Radiate; Chemo; Stem cell

Rituximab; Chemo

Print


AUTOANTIBODIES AND IMMUNE POLYNEUROPATHIES

Introduction

  • Serum factors associated with immune-mediated polyneuropathies
    • Monoclonal antibodies (M-proteins) without identifiable antigenic targets
    • Cryoglobulins
    • Monoclonal and polyclonal autoantibodies that bind to specific neural components
  • Methodology: Optimal techniques increase the sensitivity & specificity of antibody detection.
  • Clinical correlations
    • Serum autoantibodies
      • Are markers for immune-mediated, treatable disorders
        • Provide diagnostic information
        • Suggest adequacy of treatment: When titers reduced more than 60%
      • Suggest specific avenues of therapy
    • Many IgM autoantibody-associated neuropathies
      • Are difficult to treat with the more commonly used T-cell related immunosuppressants, including
      • May improve after administration of IVIg or cytotoxic agents, such as

 

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