Laboratory Markers of Toxins and Detoxification

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Laboratory Markers of Toxins and Detoxification

• Dr. Eve Bralley and
• Dr. Elizabeth Redmond

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Introduction

• Humans are constantly exposed to exogenous and
  endogenous toxins.
• Detoxification is the process of transforming and
  removing xenobiotics.
• Biotransformation occurs in steps I and II
• Detoxification and biotransformation have their
  own energy, nutrient and regulatory requirements

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Classes of Toxins and Methods of Detoxification
Figure 1
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http//www.alternative-therapies.com/at/web_pdfs/i
fm_proceedings_low.pdf
The Proceedings From the 13th International
Symposium of The Institute for Functional
Medicine Managing Biotransformation The
Metabolic, Genomic, and Detoxification Balance
Points

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Detection of Toxic Burden

• Ammonia burden
• Citrate, isocitrate, cis-aconitate
• Orotate
• 8-OH-dG
• p-hydroxyphenyllactate
• 2-Methylhippurate
• Dysbiosis markers

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Additional Markers of Toxicant Exposure

• Urinary glucarate
• Phase I and II upregulation
• herbicide, fungicide, petrochemical, alcohol and
  drug exposure
• Urinary benzoate
• Glycine conjugation

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Detection of Toxic Burden

• Porphyrins
• Heavy Metals
• Whole blood, RBC, serum
• Urine
• Hair

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Phases of Detoxification

• Phase I
• primarily cytochromes
• addition of functional groups
• Phase II
• conjugation reactions
• attaches a water-soluble moiety
• Phase III
• antiporter system

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Antioxidants Phytonutrients Vitamin C, E Minerals
Phase II Nutrients Glycine NAC B vitamins
Phase I Nutrients B vitamins Glutathione
Flavanoids
Phase II (Conjugation Pathways) Sulfation Amino
Acid conjugation Glucronidation Glutathione
Metnhylation
Phase I (Cytochrome P450) Reactions Oxidation
Excretion Blood Urine Feces
Toxins
Intermediate Metabolite
ROS
Free Radicals
Cell damage
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Phase I

• Operates via Cytochrome P450 enzymes (Cyp)
• Bioactivation adds an attachment site to the
  toxin - Requires NADH (from niacin)
• Can result in a more toxic toxin
• Generates reactive oxygen species thus ensuring
  antioxidant status is important

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Phase I - Cyp Liver Enzymes
P450 Enzyme Percent of Total Substrates
Cyp3A4,5 29 Testosterone, cyclosporine
Cyp2C8,9,18 18 Warfarine, R-mephenytoin
Cyp1A2 13 Cafeine, aflatoxin B1
Cyp2E1 6 Ethanol, Carbon tetrachloride
Cyp2A6 4 Coumarin
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Phase II

• Products of phase I detoxification are further
  metabolized
• Water soluble molecules are added creating
  inactive products called conjugates that are
  excreted in bile and urine
• Conjugases
• Sulfation
• Glucuronidation
• Amino acid conjugation
• glycination
• Glutathione conjugation
• Methylation
• Acetylation

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Glutathione and Sulfate Conjugate Formation
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Phase I and II Enzyme Induction

• Mono-functional inducers
• These elevate Phase II enzymes (such as
  glutathione S-transferases, NAD(P)Hquinone
  reductase, UDP-glucuronosyl-transferases) in
  various tissues without significantly raising the
  Phase I enzyme
• Examples cigarettes, charred meats,
  glucocorticoids,
• Bi-functional inducers
• These induce both Phase I and Phase II enzymes
  
• of xenobiotic metabolism.
• Examples flavonoids, garlic, rosemary, soy,
  cabbage, polycyclic aromatic hydrocarbons

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Use of Challenge Compounds to Evaluate Phases of
Detoxification
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Caffeine Clearance

• 200 mg dose
• 2 and 8 hour saliva collection
• Phase I microsomal P450 mixed function oxidase
  activity

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Caffeine Clearance

• If High
• Liver is actively removing caffeine
• Enzyme induction
• Minimize exposure to environmental toxins
• If Low
• Genetic polymorphisms or loss of liver function
• Processing and degradation of foreign compounds
  is slower than normal
• Xenobiotics could accumulate in adipose tissue
• Drugs cimetidine, amphetamines, grapefruit
  (narengenin)
• Low exposure to toxins

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Acetominophen Biotransformation

• 650 mg dose
• Urinary acetaminophen sulfate and acetaminophen
  glucuronide
• Sulfate and B-vitamin status

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Aspirin Biotransformation

• 650 mg dose
• O-hydroxyhippurate
• Glycine, pantothenic acid status

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Conjugation Pathways Used for Specific Compounds
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Phase I/II Ratios

• Bioactivation
• Impairment in phase II conjugation could lead to
  accumulation of toxic intermediates
• Supplement with antioxidant nutrients
• Free radicals
• Cell injury
• DNA modification
• Haptens

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Factors that influence detoxification

• Age
• Gender
• Diet and Lifestyle
• Environment
• Disease State
• Genetic polymorphisms
• Supplement use
• Intestinal Health

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Supporting Detoxification

• Co-factors and Nutrients
• Nicain, B6, glycine, NAC
• Antioxidants/protective nutrients
• vitamin C and E, zinc, lipoic acid
• Phytonutrients can trigger the activity of gene
  response elements
• Sulforaphane from broccolli

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Polymorphysims

• There are many known polymorphysims that can
  affect detoxification.
• Example of effects
• Felton, J.S. What do diet-induced changes in
  phase I and phase II enzymes tell us about
  prevetnion from exposure to heterocyclic amines?
  Journal of Nutrition, Oct 2006. 136(10) 2683-4S
• http//jn.nutrition.org/cgi/content/full/136/10
  /2683S

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Case 1

• Case 1
• 52 year old man
• Suffers from occasional abdominal cramps,
  fatigue, and malaise.

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Case 1
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Case 2

• 50 year old female
• Fatigue, depression, joint pain, IBS symptoms

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Detoxification Capacity Profile
Case 2
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Amino Acid Profile 20 - Plasma
Case 2
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OrganixSM Profile
GIfx
Case 2
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Case 3

• 49 year old male
• Depression, fatigue, headaches, overweight
• Moderate alcohol intake

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Detoxification Capacity Profile
Case 3
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Amino Acid Profile 20 - Plasma
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Elements Erythrocytes/Whole Blood
OrganixSM Profile
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Case 4

• 41 year old, painter
• Environmentally sensitive

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Detoxification Capacity Profile
Case 4
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OrganixSM Profile
GIfx
Case 4
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Thank You

• For questions on these or any testing please go
  to www.metametrix.com or call at 800-221-4640