Dr. Dicken Weatherby's "Foundations of Functional Diagnosis" Training INSIDER S GUIDE. Dicken Weatherby, N.D.

Transcription

1 Dr. Dicken Weatherby's "Foundations of Functional Diagnosis" Training INSIDER S GUIDE Advanced FDM Testing: The Detoxification Profile Dicken Weatherby, N.D. Limits of Liability & Disclaimer of Warranty I have designed this book to provide information in regard to the subject matter covered. It is made available with the understanding that the author is not liable for the misconception or misuse of information provided. The purpose of this book is to educate. It is not meant to be a comprehensive source for the topic covered, and is not intended as a substitute for medical diagnosis or treatment, or intended as a substitute for medical counseling. Information contained in this book should not be construed as a claim or representation that any treatment, process or interpretation mentioned constitutes a cure, palliative, or ameliorative. The information covered is intended to supplement the practitioner s knowledge of their patient. It should be considered as adjunctive support to other diagnostic medical procedures. This material contains elements protected under International and Federal Copyright laws and treaties. Any unauthorized reprint or use of this material is prohibited. Insider s Guide Detox Profile Testing Page 1 of 26

2 Contents Contents... 2 Introduction... 5 Background... 5 The Detoxification Capacity Profile Test... 5 Caffeine... 6 Acetaminophen... 6 Aspirin... 6 Directions... 7 Things to tell patient before they take this Test:... 7 Results... 7 Phase 1 Detoxification Results... 8 Low caffeine clearance (Phase I)... 8 Suspect... 8 Consider the following actions... 9 Phase 1 Inhibitors... 9 High caffeine clearance... 9 Phase 1 Inducers Consider the following actions Phase 1 Detoxification May Exhaust Critical Nutrients The key nutrients needed for phase 1 detoxification are Phase II Detoxification Results Glutathione Conjugation If low acetaminophen mercapturate (glutathione conjugation) Possible causes Consider the following actions Glycine Conjugation If low salicyluric acid (glycine conjugation) Possible causes Consider the following actions Sulfation Conjugation If low acetaminophen sulfate Suspect insufficient sulfation Possible causes Insider s Guide Detox Profile Testing Page 2 of 26

3 Consider the following actions Influences on sulfation Glucuronidation Conjugation If low acetaminophen glucuronide Possible causes Consider the following actions Cysteine and Sulphate If high or normal plasma cysteine with low plasma sulfate Possible causes Consider the following actions If high plasma cysteine with normal plasma sulfate Suspect cysteine accumulation Possible causes Consider the following actions Cysteine/Sulphate Ratio Normal Cysteine/Sulphate Ratio Elevated Cysteine/Sulphate Ratio Phase 2 Conjugation Requirements Phase 2 Nutrients Phase 2 Inhibitors Phase 2 Inducers Who does this test? The Comprehensive Detox Profile Includes an Oxidative Stress Profile Background Results Catechol and/or 2,3 DHB If high catechol and/or 2,3 DHB: Suspect increased hydroxyl radical activity Possible causes Consider the following actions If high lipid peroxides Suspect increased hydroxyl radical activity Possible causes Consider the following actions Reduced Glutathione, Glutathione Peroxide, and Superoxide Dismutase Insider s Guide Detox Profile Testing Page 3 of 26

4 If low reduced glutathione Suspect a depleted glutathione reserves Possible causes Consider the following actions If low superoxide dismutase (SOD) and/or glutathione peroxidase (GSH-Px) 26 Suspect that there s a depleted endogenous antioxidant reserves Possible causes Consider the following actions Resources Insider s Guide Detox Profile Testing Page 4 of 26

5 Introduction The graphics in this section are taken from the Genova Diagnostic s report. Background This test looks at the key detoxification pathways in phase 1 and phase 2 detoxification. All exogenous and endogenous toxin and chemical passes through a 2-phase liver detoxification process, which we covered in depth in the physiology of detoxification section. Here s a brief summary: Phase I involves a series of enzymatic processes collectively called the cytochrome P450 enzyme complex. Here the toxin is biotransformed through oxidation, reduction or hydrolysis. Phase 1 takes a toxin and renders it more soluble and prepares it for the Phase 2 process. One of the problems with the phase 1 process is that it tends to produce high levels of reactive oxygen species (ROS), free radicals and oxidative stress. Phase 2 involves a series of enzymatic processes to make the toxin soluble so it can be excreted in the feces or the urine. The liver does this by adding a hydrophilic molecule to the metabolite or toxin. This is especially important for lipophillic toxins or metabolites. The phase 2 pathways include glutathione, sulfation/sulfoxidation, glycine, acetylation, methylation and glucuronidation. Individual toxins pass from the phase 1 pathway and typically enter one or more of the 6 phase 2 pathways. The Detoxification Capacity Profile Test A definitive assessment of the detoxification function can be made using the Comprehensive Detoxification Profile from Genova Diagnostics (other labs have similar tests). This test uses common substances to challenge the liver's detoxification ability using urine and saliva samples. Research into the detoxification pathways show that acetaminophen, caffeine, and aspirin are detoxified by specific pathways. These 3 substances are the challenge molecules that will reveal weaknesses and blockages in your patient s detoxification system. The following picture is taken from Genova Diagnostic and shows the various pathways the 3 test substances follow and the various metabolites that are measured in the urine and the saliva. Insider s Guide Detox Profile Testing Page 5 of 26

6 Used with permission from Genova Diagnostics. all rights reserved Genova Diagnostics Caffeine Caffeine allows us to test a number of P-450 systems simultaneously. We know how much is ingested and know how much is collected in the urine so we are actually measuring caffeine clearance. The caffeine clearance gives us a reading of the function of the P450 complex. Acetaminophen Acetaminophen also passes through the phase 1 pathway. Acetaminophen is converted in phase 1 into a measurable toxic metabolite intermediate called NADPQ. NADPQ is detoxified in phase 2 via the glutathione pathway. Acetaminophen also passes through the sulfation and the glucuronidation pathway and this test measures the metabolites from each of these pathways giving us a way to measure the activity of these pathways. Aspirin Aspirin breaks down into a molecule called salicylic acid (salicylate), which is detoxified primarily through the glycination pathway. A number of other environmental toxins and xenobiotics are detoxified by this pathway so the body may not have sufficient reserves of glycine to detoxify the salicylate properly. Salicylate is also detoxified via glucuronidation. The metabolism of salicylate also produces three oxidized derivatives, which give us an insight into the free radical status of the body. Insider s Guide Detox Profile Testing Page 6 of 26

7 Directions The following directions are for the Comprehensive Detoxification Profile from Genova Diagnostics In the Detoxification Profile, one caffeine caplet (200 mg) is taken in the morning and its clearance is assessed from two salivary specimens collected two and eight hours after ingestion. Aspirin and acetaminophen are ingested in the evening and the products of detoxifying reactions are assessed in a 10-hour overnight urine specimen. The challenge dose consists of two capsules of aspirin (650 mg total) and two capsules of acetaminophen (650 mg total). The only side effect is potential drowsiness. In the Comprehensive version of this profile, the urine specimen is analyzed for levels of lipid peroxides. In addition, glutathione, glutathione peroxidase, superoxide dismutase, cysteine, and sulfate are assessed from fasting blood specimens. Things to tell patient before they take this Test: Results Patient must discontinue aspirin and acetaminophen (for 24 hours) They must avoid alcohol, caffeine, fruits, nuts, spices, and seasonings They should fast 10 hours overnight (during urine collection and before blood draw) You should arrange a blood draw on the morning of the urine collection if you are doing the Comprehensive Detoxification profile, which includes the oxidative stress assessment The following illustration is taken from the Comprehensive Detoxification Profile report from Genova Diagnostics Insider s Guide Detox Profile Testing Page 7 of 26

8 Used with permission from Genova Diagnostics. all rights reserved Genova Diagnostics Phase 1 Detoxification Results Used with permission from Genova Diagnostics. all rights reserved Genova Diagnostics This test measures caffeine clearance, which gives us a way of measuring liver cytochrome P450 activity. Low caffeine clearance (Phase I) A low caffeine clearance indicates that the activity of the cytochrome p450 enzyme complex is slow, which makes metabolic detoxification difficult. There are certain medications that will inhibit p450 activity these include amphetamines, cimetidine, and oral contraceptives (see the complete list below). Suspect Insider s Guide Detox Profile Testing Page 8 of 26

9 Impaired Phase 1 detoxification activity Exposure to P450 inhibitors (see the complete list below). Sluggish hepatic function Underlying liver disease/hepatic damage from drugs or alcohol Insufficiencies of nutrient cofactors for P450 (thiamin, riboflavin, niacin, Mg, Fe, Mo) Diet rich in saturated and hydrogenated fats, deficient in essential fatty acids Hypothyroidism Lack of exposure to chemicals (rare) Consider the following actions Eliminate any inhibiting substances Consider nutritional and/or herbal hepatic support Lipotropic factors, phosphatidylcholine, taurine, acetyl-l-carnitine, silymarin, taraxacum, chelidonium, chionanthus, catechin, essential fatty acids, antioxidants, etc. Correct any nutrient deficiencies Treat underlying liver disease or hypothyroidism Provide nutrition for Phase 2 and antioxidants, to prepare for the possibility of an up-regulated phase 1 enzyme system. Phase 1 Inhibitors Many substances (phase 1 inhibitors) can cause the phase 1 process to slow down. 1. Medications: anti-ulcer drugs (cimetidine), benzodiazapines (valium), antihistamines, anti-fungal agents (ketoconazole and other -azoles), oral contraceptives, omeprazole, amphetamines, SSRI antidepressants, erythromycin, clarithromycin 2. Heavy metals: mercury, lead and cadmium 3. Diet: high levels of sugar, trans fatty acids (partially hydrogenated fats) 4. Foods and spices: Grapefruit (naringinin), turmeric (curcumin), cloves (eugenol), cayenne pepper (capsaicin) 5. Toxic compounds from the gut- the organic acids produced by pathogenic bacteria and the putrification of proteins and the fermentation of carbs 6. Others Quercitin, Iron deficiency High caffeine clearance High caffeine clearance reflects that the cytochrome p450 enzyme complex is being induced. This may be due to due to toxin exposure (see the list below for possible phase 1 inducers). A high caffeine clearance also implies an increase production of free radicals; also implies greater production of free radicals. Insider s Guide Detox Profile Testing Page 9 of 26

10 Suspect an induced cytochrome P450 enzyme system. Phase 1 Inducers Substances (phase 1 inducers) can cause phase 1 to work faster than phase 2. The following substances activate phase 1 detoxification 1. Drugs: alcohol, nicotine, Phenobarbital, steroids, sulfonamides, barbiturates 2. Foods: Brassica family i.e. cruciferous vegetables (high in indol), char-broiled meat, high protein diets, citrus fruits (not grapefruit) 3. Vitamins: B1, B3 and C 4. Environmental toxins (xenobiotics): carbon tetrachloride, exhaust fumes, paint fumes, dioxin, pesticides 5. Herbs: Caraway, dill seed 6. Others: Tobacco smoke, alcohol, gut derived toxins, intestinal hyperpermeability, charcoal-broiled foods Consider the following actions Purify home and work environment Consider the Intestinal Permeability test and a Digestive/Microbial Stool Test to rule out gut-derived toxins Increase intake of antioxidants to deal with the increase in free radicals and oxidative stress Nutritionally support Phase 2 pathways to prevent buildup of toxic intermediate metabolites Remember, stimulation of phase 1 with nutrients and non-toxic stimulants is a good idea. However, it becomes a problem when phase 2 cannot keep up. Phase 1 Detoxification May Exhaust Critical Nutrients High exposure to toxins in phase 1 may exhaust the supply of critical nutrients needed for phase 2. High levels of toxin exposure in phase 1 can deplete nutrients that are also used in phase 2. For instance, glutathione stores for the glutathione dependent processes in Phase 2 can be used up in phase 1, causing a build-up of dangerous toxins in the body. The key nutrients needed for phase 1 detoxification are B vitamins: B2, B3, B6, B12 and folic acid Glutathione BCAA- L-leucine, L-isoleucine and L-valine Flavanoids Minerals: Copper, Zinc, and magnesium Vitamin C Insider s Guide Detox Profile Testing Page 10 of 26

11 Phase II Detoxification Results The Phase II conjugation pathways tested with the Comprehensive Detox Profile include: Glutathione Conjugation Glycine Conjugation Glucuronide Conjugation Ratio of Cysteine to Sulfate Glutathione Conjugation The body uses glutathione to detoxify many fat-soluble toxins and carcinogens making them water-soluble. About 60% of the toxins excreted in the bile are detoxified via glutathione conjugation. Glutathione circulates in the blood acting as a free radical quencher. It can grab onto hundreds of different environmental chemicals, drag them out of the blood into the liver, then to the GB and into the gut where they will be excreted in the feces. It s essential to make sure that your patients are not constipated when doing Detox work with them. Used with permission from Genova Diagnostics. all rights reserved Genova Diagnostics Glutathione reserves can be run-down quickly, causing deficiency. For every molecule that s detoxified, your body uses use up, depletes, or loses forever a molecule of glutathione plus a molecule of ATP. This is one of the reasons we will be recommending that reduced glutathione be part of your detoxification protocols. This portion of the test measures the percentage recovery of Acetaminophen Mercapturate If low acetaminophen mercapturate (glutathione conjugation) Suspect insufficient glutathione conjugation, which makes it difficult for the removal of toxins from the body. Possible causes A depletion of reduced glutathione in the body Exposure to high levels of toxic exhaust Excessive exercise, which causes oxidative stress from increased free radical production Alcohol, which can block glutathione production Insider s Guide Detox Profile Testing Page 11 of 26

12 Impairment in the other Phase 2 pathways, resulting in the compensation by the glutathione conjugation system Excess exposure to xenobiotics and/or free radical production Inadequate nutrient precursors (cysteine, glutamic acid and glycine) Depletion of reduced glutathione Insufficient nutrient cofactors Induced P450 activity (more compounds to detoxify) Genetic uniqueness Enhanced bile production (mercapturate elimination via the bile) Consider the following actions Identify and reduce sources of xenobiotic exposure and free radical generation Increase intake of Glutathione and its precursors Reduced glutathione (one of the best sources is Recancostat from Integrative Therapeutics), N-acetylcysteine, glycine, L-methionine, L- glutamine Address impairments in other Phase 2 pathways Address induced P450 system, to reduce burden on GSH conjugation Increase intake of nutrient cofactors: Zn, Cu, riboflavin, niacin, selenium, Mg, vitamin B6, B12, folic acid Diet: Increase intake of cruciferous vegetables (induces conjugating enzyme) Glutathione rich foods include asparagus, avocado, walnuts, cabbage, broccoli, Brussels sprouts. Dietary glutathione appears to be well absorbed. The same cannot be said of supplemental glutathione unless in the reduced form. Insider s Guide Detox Profile Testing Page 12 of 26

13 Glycine Conjugation Glycine conjugation is part of the amino acid conjugation system. The body uses five amino acids for detoxification: 1. Glycine, 2. Taurine, 3. Glutamine, 4. Arginine, 5. Ornithine. All 5 amino acids can be synthesized by the body. Glycine is the most important amino acid used for toxin biotransformation. Used with permission from Genova Diagnostics. all rights reserved Genova Diagnostics This portion of the test measures the percentage recovery of salicyluric acid If low salicyluric acid (glycine conjugation) Suspect insufficient glycine conjugation which makes it difficult for the removal of toxins from the body. Possible causes Insufficient glycine available Insufficient nutrient cofactors Underlying hepatic disease Genetic uniqueness Consider the following actions Increase intake of glycine Increase intake of nutrient cofactors Cysteine, pantothenic acid, Mg (coenzyme A required for activation of the metabolite to be conjugated) Increase alkaline ash foods to enhance glycination Insider s Guide Detox Profile Testing Page 13 of 26

14 Sulfation Conjugation Sulfation uses sulfur containing compounds to detoxify toxins and make them watersoluble. It is one of the weakest phase 2 pathways due to inadequate supply of sulfur from diet. Sulfation is responsible for the transformation of the following: Hormones (steroid and thyroid) Drugs Industrial chemicals Phenol containing compounds (plastic, disinfectants) Toxins from the intestines Neurotransmitters Used with permission from Genova Diagnostics. all rights reserved Genova Diagnostics Acetaminophen (also known as paracetamol) is an example of a molecule detoxified primarily through the sulfation pathway. It is the test molecule for the sulfation pathway in this test. Many people have poor sulfation and will have a decreased acetaminophen sulfate conjugation. These people are most susceptible to environmental illness and problems of the nervous system, including Parkinson s disease and motor neuron disease. Weak sulfation has also been associated with rheumatoid arthritis, biliary cirrhosis, food and multiple chemical sensitivities. This portion of the test measures the percentage recovery of acetaminophen sulfate If low acetaminophen sulfate Suspect insufficient sulfation Possible causes Depletion of sulfate Excess exposure to xenobiotics or free radical production Inadequate dietary sulfate available Insufficient nutrient cofactors Induced P450 activity Impaired sulfoxidation ability (especially if high cysteine/sulfate ratio, with low plasma sulfate) Molybdenum insufficiency, especially if low plasma sulfate Molybdenum or vitamin B6 excess (can inhibit sulfation) Insider s Guide Detox Profile Testing Page 14 of 26

15 Underlying hepatic disease Genetic uniqueness Sulfation may be low if a shared pathway (e.g. glucuronidation) is taking on more compound than usual Consider the following actions Rule out excess xenobiotic exposure, especially if elevated caffeine clearance Consider foods or supplements containing sulfate precursors, unless high cysteine/sulfate ratio with low plasma sulfate L-methionine, L-cysteine, N-acetylcysteine, Reduced glutathione Increase intake of nutrient cofactors Zn, Cu, riboflavin, selenium, Mg, vitamin B6, B12, folic acid Consider inorganic sulfate (especially if high cysteine/sulfate ratio with low plasma sulfate) Increase intake of molybdenum, if signs of insufficiency such as impaired sulfoxidation and/or sulfite toxicity Avoid vitamin B6 or Mo supplementation, if suspect excess Influences on sulfation A diet low in sulfur has been shown to reduce sulfation Sulfation can be induced via the following: 1. Supplemental sulfur (taurine, cysteine and methionine) 2. Sulfur-containing foods (garlic, egg yolks, onions, broccoli and red peppers) Sulfation may also be reduced by excessive molybdenum or vitamin B6 (over 100mg/day) Insider s Guide Detox Profile Testing Page 15 of 26

16 Glucuronidation Conjugation Glucuronidation is catalyzed by glucuronysyltransferase enzymes in the liver. This enzyme complex is susceptible to genetic polymorphisms: People with problems with this enzyme are more susceptible to Gilbert s syndrome and reduced glucuronidation making it difficult for them to excrete various hormonal endo and exo toxins. Glucuronidation detoxifies the following: Bilirubin Steroid and thyroid hormones Retinoids Bile acids Lipophilic xenobiotics Used with permission from Genova Diagnostics. all rights reserved Genova Diagnostics People suffering from oxidative stress may have diminished capacity for Phase II glucuronide conjugation. This portion of the test measures the percentage recovery of acetaminophen glucuronide If low acetaminophen glucuronide Suspect impaired glucuronidation detoxification Possible causes Insufficient carbohydrate reserves (e.g. fasting or insulin insensitivity) Possible free radical damage to mitochondria (ATP necessary for glucuronidation) Iron deficiency Excess vitamin K Insufficient nutrient cofactors for glucuronidation (calcium d-glucarate) Hypothyroidism (delays maturation of conjugating enzyme) Antibiotics chloramphenicol and novobiocin can interfere with the conjugating enzyme Genetic uniqueness (e.g. Gilbert's disease) Glucuronidation may be low if a shared pathway (e.g. sulfation) is taking on more compound than usual Consider the following actions Improve glucose utilization or insulin resistance, if relevant Insider s Guide Detox Profile Testing Page 16 of 26

17 Supplement antioxidants if evidence of free radical damage (you will see the oxidative stress on this test as well) Correct nutrient deficiencies Increase intake of nutrient cofactors - Diindolylmethane (DIM) (Allergy Research, Integrative Therapeutics or other companies) mg twice/day and Calcium d-glucarate (Integrative therapeutics 360 mg bid, Biotics Research) L-glutamine, aspartic acid, niacin, vitamin B6, Fe, Mg Rule out hypothyroidism Support other Phase II pathways, especially sulfation and glycination, to reduce burden on glucuronidation Increase intake of cruciferous vegetables (broccoli, brussels sprouts induce conjugating enzyme) Probiotics (prevent the increase of beta glucuronidase enzyme in the gut from dysbiotic bacteria which strips the glucuronide molecule from the toxin it is escorting out of the body) Cysteine and Sulphate Used with permission from Genova Diagnostics. all rights reserved Genova Diagnostics If high or normal plasma cysteine with low plasma sulfate Insider s Guide Detox Profile Testing Page 17 of 26

18 Suspect impaired sulfoxidation ability (cysteine -> sulfite -> sulfate) Possible causes Genetic enzyme defect Molybdenum insufficiency (Mo needed for the conversion of sulfite to sulfate) Consider the following actions Supplement with inorganic sulfate to bypass sulfoxidation Na sulfate, Mg sulfate, etc. (Note: supplementation with N-acetylcysteine, methionine or glutathione may exacerbate symptoms.) Supplement with molybdenum if suspect insufficiency (e.g. sensitivity to sulfites) If high plasma cysteine with normal plasma sulfate Suspect cysteine accumulation Possible causes o Excess dietary protein or supplementation o Excess catabolism (e.g. muscle wasting), o A block in the conversion of cysteine to glutathione, which is magnesium dependent o A block in conversion of cysteine to Taurine, which is B6 dependent Consider the following actions o Address any cause of muscle wasting o Supplement with Mg and/or vitamin B6 o Supplement with Reduced glutathione (Recancostat) and/or taurine to bypass possible blocks Cysteine/Sulphate Ratio Used with permission from Genova Diagnostics. all rights reserved Genova Diagnostics Normal Cysteine/Sulphate Ratio Insider s Guide Detox Profile Testing Page 18 of 26

19 If the plasma cysteine, plasma sulfate and the cysteine/sulfate ratios are all within the reference range the body is adequately generating inorganic sulfate from cysteine and the sulfoxidation pathway is operating properly. Elevated Cysteine/Sulphate Ratio It appears that patients with neurological problems such as Alzheimer s disease, Parkinson s disease, and motor neuron diseases are deficient in the enzyme that catalyzes the conversion of cysteine to sulfate via sulfoxidation. This causes an elevated plasma cysteine/sulfate ratio. Patients who are deficient in sulfate and have a high xenobiotic load will benefit from cysteine and glutathione supplementation. However those with an elevated plasma cysteine/sulfate ratio have a problem with the sulfoxidation step and may worsen with cysteine supplementation because the body is unable to convert the cysteine to sulfate. These patients may require inorganic sulfate as a supplement, in order to bypass this sulfoxidation step. They may also require molybdenum supplementation because molybdenum is a cofactor for sulfite oxidase, one of the enzymes involved in sulfoxidation. Molybdenum status is important in the assessment of poor sulfation capability. Look at uric acid levels in the blood as a gateway test for assessing molybdenum status. The enzyme xanthine oxidase, which is responsible for the formation of uric acid in the body, is molybdenum dependent. Low uric acid levels may be caused by low molybdenum levels (other causes of a decreased uric acid include B12 and folate deficiency, copper deficiency, prolonged use of aspirin, corticosteroids, heavy metals toxicity). Phase 2 Conjugation Requirements Phase 2 Nutrients Phase 2 detoxification requires key nutrients for the activation of the specific detoxification enzyme complexes. It also requires energy to function and synthesize conjugating molecules. Without nutrients and energy, phase 2 detoxification can slow down, leading to increased toxin build-up NUTRIENTS Glutathione Amino acids Glutathione, B6, C, precursors (Cysteine, Glycine, Glutamic Acid, and co-factors), EFAs (GLA, Flax Seed Oil, EPA) Glycine Methylation S-adenosyline-methionine (SAM-e), Mg, Folic Acid, B-12, Methyl Donors Sulfation/ Sulfoxidation Cysteine, methionine, molybdenum, MSM, Cofactors (B-12, Folic Acid, Methyl Donors, Magnesium, B-6/P-5-P) Insider s Guide Detox Profile Testing Page 19 of 26

20 Acetylation Acetyl- CoA, Molybdenum, Iron, Niacinamide, B2 Glucoronidation Calcium d-glucarate, glucaric acid, Mg There are also substances that inhibit phase 2 (phase 2 inhibitors), and substances that induce phase 2. Phase 2 Inhibitors Phase 2 system Glutathione Inhibitors Deficiencies of: Selenium, B2, zinc, glutathione Amino acids Low protein diet Methylation Deficiencies: folic acid and vitamin B12 Sulfation/ Sulfoxidation Acetylation NSAIDS (aspirin), tartrazine (yellow food coloring), molybdenum deficiency Deficiencies of: vitamin B2, B5 or C Glucoronidation Aspirin, probenecid Phase 2 Inducers Phase 2 system Glutathione Amino acids Inducers Brassica-family foods (cabbage, broccoli, Brussels sprouts), limonene (citrus peel, dill and caraway) Glycine Methylation Sulfation Lipotropic nutrients (choline, betaine, methionine, folic acid, vitamin B12) Cysteine, taurine and methionine Insider s Guide Detox Profile Testing Page 20 of 26

21 Acetylation None known Glucoronidation Fish oils, cigarettes, Phenobarbital, limonene (citrus peel, dill and caraway), oral contraceptives Who does this test? This is a short list of some of the well known labs who do this test. Genova Diagnostics Comprehensive Detoxification Profile Metametrix - Detoxification Capacity Doctor s Data - Hepatic Detox Profile The Comprehensive Detox Profile Includes an Oxidative Stress Profile I know we have already gone over this test but we feel a review is warranted because the oxidative stress panel is part of the comprehensive Detox profile. The graphics in this section are taken from the Genova Diagnostic s report. Background The oxidative stress profile is part of the Comprehensive Detoxification Profile. We ll go through the test from Genova Diagnostics here. It can be ordered as a stand alone test and a number of other companies run Oxidative Stress Panels too. The Oxidative Stress Profile includes the measurement of the following: 1. Two derivatives of salicylate: catechol and 2, 3-dihydroxybenzoate (2,3-DHB) 2. Urine lipid peroxides 3. Blood glutathione 4. Glutathione peroxidase (GSH-Px) 5. Superoxide dismutase (SOD). Insider s Guide Detox Profile Testing Page 21 of 26

22 Used with permission from Genova Diagnostics. all rights reserved Genova Diagnostics Catechol and 2,3-DHB These are direct products of hydroxyl radical attack upon salicylic acid. The amount of 2,3-DHB appearing in the urine after an aspirin challenge appears to be a direct reflection of hydroxyl radical concentrations. Catechol correlates with 2,3-DHB as an indicator of oxidative stress. Glutathione- This is a measure of whole blood glutathione and is a reflection of the body s reserve. Glutathione acts as an antioxidant and as a critical nutrient in the phase 2 detoxification pathway. It is also a minor donor of sulfate used in detoxification reactions. This test measures the glutathione in a fasting blood sample and it s a direct measure of the functional reserves after an acetaminophen challenge. It assesses the functional reserves after the detoxification capacity has been tested. Glutathione Peroxidase (GSH-Px) - Glutathione peroxidase is a selenium-dependant enzyme found primarily in the cytoplasm and to some extent the mitochondria. Each molecule of GSH-Px has 4 atoms of selenium. It acts as a lipid peroxide scavenger in the cytoplasm and the mitochondria. Optimal levels of this enzyme prevent the build up of lipid peroxides in this tissue. Superoxide Dismutase (SOD) SOD is found in a number of different tissues including the cytosol and the mitochondria. SOD is dependent on various minerals depending on the tissue it is found in. In the cytosol SOD is dependent on zinc and copper. In the mitochondria it is dependent on manganese. This enzyme catalyzes the reaction that converts the superoxide radical in to the less reactive H 2 O 2 form. The mitochondria generate sufficient amounts of superoxide radical that adequate levels of SOD and GSH-Px are needed to protect this organelle from oxidative damage. This is especially important during times of high energy need, which would include excessive muscular activity (sports, running, body building etc.). SOD levels are also Insider s Guide Detox Profile Testing Page 22 of 26

23 high in dysfunction and diseases of the muscle, sclerosis, myositis and malignant melanoma. Increased levels of SOD may also be an indicator of pesticide exposure because SOD is a sensitive marker for exposure to pesticides. SOD are often decreased in people with early onset of diabetes and glucose intolerance, inflammatory conditions, and in zinc deficiency. Urine Lipid Peroxides and Hydroxyl Radical Markers - Polyunsaturated fatty acids (PUFAs) are susceptible to hydroxyl radical attacks that produce lipid peroxides. Lipid peroxides can be measured in the urine and elevated levels are suggestive of hydroxyl radical activity and reflect oxidative damage occurring in the body. Essential Fatty Acids are typically bound into every cell membrane and are especially vulnerable to attacks from hydroxyl radical activity and oxidative stress. This can cause a deficiency in essential fatty acids, especially those that are highly unsaturated (EPA and DHA, which have 5 and 6 points of unsaturation respectively). Results Catechol and/or 2,3 DHB Used with permission from Genova Diagnostics. all rights reserved Genova Diagnostics If high catechol and/or 2,3 DHB: Suspect increased hydroxyl radical activity Possible causes Excess exposure to xenobiotics or gut-derived toxins/upregulated cytochrome P450 activity Other sources of free radicals: Eg. Inflammation, infection, intestinal dysbiosis, trauma, radiation, ischemia Inadequate nutritional antioxidant reserves Iron overload (Can induce hydroxyl radical production) Consider the following actions Identify and reduce exposure to toxic substances and other sources of free radicals Insider s Guide Detox Profile Testing Page 23 of 26

24 Consider running a Detoxification Profile (this may have already been done) Look into intestinal hyperpermeability, bowel toxemia, and dysbiosis. Consider running the Digestive and Microbe Stool Test Check into nutrient deficiencies and heavy metals by running the RBC Erythrocyte test and/or the Heavy Metal provocation test Consider increasing intake of antioxidants: Ascorbic acid, bioflavonoids, carotenoids, tocopherols, coenzyme Q10, melatonin, lipoic acid, N- acetylcysteine Consider herbal antioxidants: Milk thistle, catechin, ginkgo biloba, licorice, hawthorne, reishi, anthocyanidins, curcumin Consider serum ferritin test to rule out excess iron, which can be a big cause of oxidative stress in the body If high lipid peroxides Suspect increased hydroxyl radical activity Possible causes Excess exposure to xenobiotics or gut-derived toxins/upregulated cytochrome P450 activity Other sources of free radicals: Eg. Inflammation, infection, intestinal dysbiosis, trauma, radiation, ischemia Inadequate nutritional antioxidant reserves Consider the following actions Identify and reduce exposure to toxic substances or other sources of free radicals Consider running a Detoxification Profile (this may have already been done) Look into intestinal hyperpermeability, bowel toxemia, and dysbiosis. Consider running the Digestive and Microbe Stool Test Check into nutrient deficiencies and heavy metals by running the RBC Erythrocyte test and/or the Heavy Metal provocation test Consider increasing intake of antioxidants, especially fat-soluble nutrients: Tocopherols, ascorbic acid, carotenoids, coenzyme Q10, melatonin, lipoic acid, glutathione, N-acetylcysteine Consider herbal antioxidants: Milk thistle, catechin, ginkgo biloba, licorice, hawthorne, reishi, anthocyanidins, curcumin Consider nutritional cell membrane support phosphatidyl choline (PhosChol from Nutrasal), taurine, essential fatty acids, esp. omega 3s, reduce partially hydrogenated fats Insider s Guide Detox Profile Testing Page 24 of 26

25 Consider running an essential fatty acid test Reduced Glutathione, Glutathione Peroxide, and Superoxide Dismutase Used with permission from Genova Diagnostics. all rights reserved Genova Diagnostics If low reduced glutathione Suspect a depleted glutathione reserves Possible causes Excess exposure to xenobiotics or gut-derived toxins Excess production of free radicals e.g. upregulated cytochrome P450 activity, inflammation, infection, intestinal dysbiosis, trauma, radiation, ischemia Inadequate GSH precursors/impaired methionine metabolism Insufficient nutrient cofactors for GSH production or metabolism Consider the following actions Identify and reduce exposure to toxic substances or other sources of free radicals Consider running a Detoxification Profile (this may have already been done) Look into intestinal hyperpermeability, bowel toxemia, and dysbiosis. Consider running the Digestive and Microbe Stool Test Consider ruling out amino acid deficiencies and defects in methionine metabolism by running an Amino Acid Profile Consider supplementary glutathione and glutathione precursors: Reduced glutathione (Recancostat), N-acetylcysteine, L-methionine, glycine, L- glutamine Consider nutrient cofactors for Glutathione and methionine metabolism: Vitamin B6, B12, riboflavin, niacin, vitamin C, folic acid, serine, Mg Insider s Guide Detox Profile Testing Page 25 of 26

26 If low superoxide dismutase (SOD) and/or glutathione peroxidase (GSH-Px) Suspect that there s a depleted endogenous antioxidant reserves Possible causes Excess exposure to xenobiotics or gut-derived toxins Excess free radical activity in the body Insufficient reduced glutathione. This is especially true with a low Glutathione peroxidase level Insufficient nutrient cofactors for enzyme activity Consider the following actions Consider options above for low reduced glutathione levels. This is especially true with a low Glutathione peroxidase level Consider increasing intake of nutrient cofactors: Manganese (to help with the mitochondrial SOD), copper and zinc (to help with the cytosolic SOD), selenium (remember GSH-Px has 4 selenium atoms in the molecule and is selenium dependent) Consider general antioxidant support: Ascorbic acid, bioflavonoids, carotenoids, tocopherols (specifically increases GSH-Px), coenzyme Q10, vitamin D3 (specifically increases SOD), melatonin, lipoic acid Consider herbal antioxidant support: Milk thistle, catechin, ginkgo biloba, licorice, hawthorne, reishi, anthocyanidins, curcumin Resources This is a short list of some of the well known labs who do this test. Genova Diagnostics Comprehensive Detoxification Profile and the Oxidative Stress Profile Genova Diagnostics 63 Zillicoa Street Asheville, NC Telephone: Local Telephone: Insider s Guide Detox Profile Testing Page 26 of 26