Minimizing Parental Health Risks What Parents Need to Know
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Laurette Janak Autism One May 2009 Part 1 of 2 Laurette.janak@verizon.net
Overview
Part One: the “What” What has been found? What it means? What to do about it?
Part Two: the “How” How the biochemical pathways for folate and glutathione function.
Metabolic Findings in Parents of Children with Autism
• 86 autism parents differ from 200 controls in the following:
– Higher homocysteine (Hcy) – Higher SAH (S-adenosylhomocysteine) – Lower GSH (glutathione) – Increased GSSG (oxidized glutathione)
J Autism Dev Disord. 2008 Nov;38(10):1966-75
Mothers of Autistic Children
J Autism Dev Disord. 2008 Nov;38(10):1966-75
46 Case Moms Mean Homocysteine Mean SAH Mean total GSH Mean GSSG 9.8 * 33 * 5.1 * .32 *
200 Control Moms 7.4 23 7.3 .24
* p < 0.001 Case Versus Control Mothers
Risk vs. Causation
Elevated Homocysteine
• What does elevated homocysteine mean? • It is a marker for a folic acid and/or vitamin B12 deficiency. (B-6) • What are the health implications of elevated homocysteine?
Homocysteine, folate, methylation and monoamine metabolism in depression
–J Neurol Neurosurg Pyschiatry 2000;69(2):228-32
52% of the depressed persons in this study had elevated homocysteine
Full text available at: http://www.pubmedcentral.nih.gov/picrender.fcgi?artid=1737050&blobtype=pdf
Depression
“Relatives of autistic individuals have high rates of major depression and social phobia that are not associated with the broad autism phenotype and cannot be explained by the increased stress associated with raising an autistic child.”
Am J Psychiatry 1999 Apr;156(4):557-63
Full text available at: http://ajp.psychiatryonline.org/cgi/reprint/156/4/557
Depression
• “Folic acid is a simple method of greatly improving the antidepressant action of fluoxetine…. Folic acid should be given in doses sufficient to decrease plasma homocysteine.”
– Enhancement of the antidepressant action of fluoxetine by folic acid: a randomized, placebo controlled trial
• J Affect Disord 2000 Nov;60(2):121-30
• What if the doctor does NOT screen you for homocysteine before putting you on a SSRI?
Depression Linked to Heart Disease
Depression has been shown to be an independent risk factor for coronary artery disease and vice versa.
–Arch Gen Psyschiatry 2004 Apr;61(4):369-76. –Eur Heart J. 2004 Jan;25(1):3-9. –Prostaglandins Leukot Essent Fatty Acids 2004 Apr;70(4):56,349
Cardiovascular Disease
• Elevated homocysteine is an independent risk factor for vascular disease
– Hyperhomocysteinemia: an independent risk factor for vascular disease
» N Engl J Med 1991 Apr 25;324(17):1149-55
– Plasma total homocysteine and cardiovascular and noncardiovascular mortality: the Hordaland Homocysteine Study.
» Am J Clin Nutr 2001 Jul;74(1):130-6
•
Folate and homocysteine metabolism: therapeutic targets in cardiovascular and neurodegenerative disorders.
» Curr Med Chem 2003 Oct;10(19):1923-9
Alzheimer’s Disease
• Elevated homocysteine is an independent risk factor for Alzheimer's disease.
– Neuroscience 2007 Mar 30;14593):942-54 – N Engl J Med. 2002 Feb 14;346(7):476-83. – Neuroreport. 2003 Jul 18;14(10):1391-4.
Parkinson’s Disease
• Disturbances in folate metabolism have been implicated as a risk factor for Parkinson’s.
– – – – – – – – Int J Mol Med. 2004 Mar;13(3):343-53 J Neurol Sci. 2003 Mar 15;207(1-2):19-23. Neurology. 2003 Feb 25;60(4):690-5. Trends Neurosci. 2003 Mar;26(3):137-46. Nutr Rev. 2002 Dec;60(12):410-3. J Neural Transm. 2002 Dec;109(12):1445-52. Int J Geriatr Psychiatry. 2002 Sep;17(9):859-64. J Neurochem. 2002 Jan;80(1):101-10.
Michael J. Fox
Schizophrenia and Homocysteine
• Elevated third trimester homocysteine levels have been associated with increased risk for schizophrenia in the offspring.
– Full text available at: http://archpsyc.ama-assn.org/cgi/reprint/64/1/31
• An increased occurrence of schizophrenia has been reported in parents of children with autism when compared to control parents.
– Pediatrics. 2008 May;121(5):e1357-62
• In a randomized, double-blind, placebo-controlled, crossover designed study, 42 schizophrenic patients with elevated homocysteine were treated with folic acid, B-12, and B-6
– The treatment resulted in significant clinical improvements
• Biol Psychiatry. 2006 Aug 1;60(3):265-9
Cancer
“Most families report cancer in the parents or grandparents, the most common being colon cancer.”
–Autism and vaccinations by Mary Megson, MD. •Wise Traditions Fall 2000
Increased mortality from cancer in individuals with autism.
–Causes of death in autism. •J Autism Dev Disord. 2001 Dec;31(6):569-76
Cancer
• Epidemiology studies have shown an association of reduced folate intake with an increased occurrence of certain cancers.
Cancer
Folic acid used as a chemopreventative agent has shown that results depend on the timing of the folic acid supplementation. For example:
Folic acid supplementation prior to any neoplastic lesions was shown to be protective against colon cancer. Folic acid supplementation once neoplastic lesions are present has been shown to fuel colon cancer growth.
Fibromyalgia Myth
• “Fibromyalgia is essentially a benign disease. It is not a degenerative or deforming condition, nor does it result in life-threatening complications.”
– (Patient information: Fibromyalgia from the Cleveland Clinic 6/22/06 Author Don L Goldenberg, MD published in www.uptodate.com)
FM patients have less total gray matter in their brains than controls. J Neurosci 2007 Apr 11;27(15):4004-7
–The longer they have had FM, the greater the gray matter loss
–Each year of FM = 9.5 times the gray matter loss as that seen in normal aging
Lower brain grey matter has been found to be associated with: Lower blood cell folate Higher homocysteine
Neurosci Lett. 2003 May 8;341(3):173-6.
Fibromyalgia Myth (It’s all in your head)
Fibromyalgia Myth
(It’s all in your head)
• “the results do confirm that there are real metabolic changes in painful muscles in PF patients.”
– (PF = primary fibromyalgia) – This study was done on muscle biopsies
• Arthritis and Rheumatism, Vol 29, No. 7 (July 1986)
•
Use of P-31 Magnetic Resonance Spectroscopy to Detect Metabolic Abnormalities in Muscles of Patients with Fibromyalgia.
– This study reiterates the findings of the 1986 study using MRS technology instead of muscle biopsy. – Findings: “impaired mitochondrial function”
•
Arthritis & Rheumatism vol 41, No3, March 1998 pp 406-413
Mitochondria produce energy (ATP)
• Muscle studies in FM have shown mitochondrial damage.
– – – – Joint Bone Spine. 2006 May;73(3):239-42. Ann Rheum Dis. 2004 Mar;63(3):245-51. Curr Rheumatol Rep. 2000 Apr;2(2):131-40. Z Rheumatol. 1998;57 Suppl 2:47-51.
Cristae - the site of the electron transport chain Matrix- the site of the citric acid cycle
Myth
(It’s all in your head)
Using either muscle biopsy or 31P magnetic resonance spectroscopy the following are found with respect to FM or CFS:
These values are increased:
Inorganic phosphate (Pi) Phosphodiesters (PDE) Pyruvate Lactate Glycolysis iNOS Increased sensitivity to NO induced suppression of oxidative phosphorylation. DNA fragmentation Lipid accumulation Lipofuscin Oxidative damage to lipids in muscle Activity of the antioxidant enzymes catalase, glutathione peroxidase and transferase
These values are decreased:
Phosphocreatine (PCr) ATP Phosphorylation potential (PP) Oxidative capacity (Vmax) Lactate dehydrogenase (LDH) Reduced nutritive blood flow during aerobic exercise Number of mitochondria. Altered morphology of mitochondria Intramuscular collagen Hydroxyproline (in muscle) Lower total concentration of the major amino acids of collagen than in controls (in muscle) Acetyl l-carnitine
Data I collected from multiple studies cited in Pubmed
Homocysteine and Mitochondria
• Correlation of plasma homocysteine and mitochondrial DNA content in peripheral blood in healthy women Atherosclerosis. 2001 Oct;158(2):399-405.
– A physiologic range of total plasma homocysteine (tHcy) was shown to have the following significant relationship:.
tHcy
mtDNA content
Chronic Fatigue Syndrome (CFS) and Homocysteine Is there a connection?
• Elevated homocysteine was found in cerebrospinal fluid of women who fulfilled the criteria for both FM and CFS.
– There was a significant correlation between the CSF-Hcy levels and fatigability. – Scand J Rheumatol 1997;26(4):301-7
•
Plasma homocysteine vs cerebrospinal fluid homocysteine
Parental Oxidative Stress
• Parents of children with autism were found to have:
– Significantly decreased levels of GSH (glutathione) – Significantly increased levels of GSSG (oxidized glutathione) – Significantly decreased ratio of GSH/GSSG
• J Autism Dev Disord. 2008 Nov;38(10):1966-75
• This means the parents are under oxidative stress.
Oxidative Stress and Psychiatric Disorders
• A meta-analysis indicates an association of oxidative stress in the majority of DSM-IV psychiatric disorders including: autism, Rett’s, ADHD, schizophrenia, anxiety, and mood disorders. Full text available at:
http://www.pubmedcentral.nih.gov/picrender.fcgi?artid=2330073&blobtype=pdf
• “…all these psychiatric disorders might benefit from a change to a whole-food plant-based diet.”
Oxidat ive Stress in Chr on ic F at igue Erythrocyte oxidative damage in chronic fatigue syndrome. Synd rome Arch Med Res. 2007 Jan;38(1):948.
Chronic fatigue syndrome is accompanied by an IgMrelated immune response directed against neopitopes formed by oxidative or nitrosative damage to lip ids and proteins. Neuro Endocrinol Lett. 2006 Oct;27(5):61521. Oxidative stress levels are raised in chronic fatigue syndrome and are associated with clin ical symptoms. Free Radic Biol Med. 2005 Sep 1;39(5):5849. Chronic fatigue syndrome: assessment of increased oxidative stress and altered muscle excitabil ity in response to incremental exercise. J Intern Med. 2005 Mar;257(3):299310. Elevated levels of protein carbonyls in sera of chronic fatigue syndrome patients. Mol Cell Biochem. 2003 Jun;248(12):935. Effect of natural and synthetic antioxidants in a mouse model of chronic fatigue syndrome. J Med Food. 2002 Winter;5(4):21120. Relationship between musculoskeletal symptoms and blood markers of oxidative stress in patients with chronic fatigue syndrome. Neurosci Lett. 2003 Jan 2;335(3):1514. Elevated nitric oxide/peroxynitrite mechanism for the common etiology of multiple chemical sensitivity, chronic fatigue syndrome, and posttraumatic stress disorder. Ann N Y Acad Sci. 2001 Mar;933:3239 Chronic fatigue syndrome: oxidative stress and dietary modifications. Altern Med Rev. 2001 Oct;6(5):4509 Specific oxidative alterations in vastus lateralis muscle of patients with the diagnosis of chronic fatigue syndrome. Free Radic Biol Med. 2000 Dec 15;29(12):12529. Blood parameters indicative of oxidative stress are associated with symptom expression in chronic fatigue syndrome. Redox Rep. 2000;5(1):3541.
Chronic Fatigue Syndrome
• “CFS patients have a lipid profile and oxidant biology that is consistent with cardiovascular risk….Supplementation with specific antioxidant medications might help to ameliorate symptoms and any potential cardiovascular complications of the illness.”
– Free Radical Biology & Medicine 39 (2005) 584-589
Oxidative Stress in Fibromyalgia
Total antioxidant capacity and the severity of the pain i n patients with fibromyalgia. Redox Rep. 2006;11(3):1315. Current concepts in the pathoph ysiology of fibromyalgia: the potential role of oxidative stress and nitric oxide. Rheumatol Int. 2006 May;26(7):58597. Antioxidant status, lipid peroxidation and n itric oxide in fibromyalgia: etiologic and therapeutic concerns. Rheumatol Int. 2006 May;26(7):598603. Free radicals and antioxidants in primary fibromyalgia: an oxidative stress disorder? Rheumatol Int. 2005 Apr;25(3):18890. Are advanced glycation endproductmodified proteins of pathogenetic importance in fibromyalgia? Rheumatology (Oxford). 2002 Oct;41(10):11637.
•
Oxidative stress is implicated in endometriosis.
– – – – J Soc Gynecol Investig. 2006 Sep;13(6):390-8 Reprod Biomed Online. 2006 Jul;13(1):126-34 Hum Reprod. 2005 Jul;20(7):2014-20. Fertil Steril. 2004 Oct;82 Suppl 3:1019-22.
•
High rates of autoimmune and endocrine disorders, fibromyalgia, chronic fatigue syndrome and atopic diseases among women with endometriosis: a survey analysis Hum Reprod. 2002 Oct;17(10):2715-24.
• “CONCLUSIONS: Hypothyroidism, fibromyalgia, chronic fatigue syndrome, autoimmune diseases, allergies and asthma are all significantly more common in women with endometriosis than in women in the general USA population.” – Full text online at: http://humrep.oxfordjournals.org/cgi/content/full/17/10/2715
• Mitochondrial dysfunction and molecular pathways of disease Exp Mol Pathol. 2007 Jan 17
– “A wide range of seemingly unrelated disorders, such as schizophrenia, bipolar disease, dementia, Alzheimer's disease, epilepsy, migraine headaches, strokes, neuropathic pain, Parkinson's disease, ataxia, transient ischemic attack, cardiomyopathy, coronary artery disease, chronic fatigue syndrome, fibromyalgia, retinitis pigmentosa, diabetes, hepatitis C, and primary biliary cirrhosis, have underlying pathophysiological mechanisms in common, namely reactive oxygen species (ROS) production, the accumulation of mitochondrial DNA (mtDNA) damage, resulting in mitochondrial dysfunction. Antioxidant therapies hold promise for improving mitochondrial performance.”
Homocysteine and Brain Development
• An animal model of maternal elevated homocysteine produced offspring with:
– Altered fetal brain development – Reduced learning abilities – Increased lipid peroxidation in the brain
• Indicates oxidative stress
– Increased brain DNA fragmentation – Abnormalities mitigated by using melatonin
• J Pineal Res. 2008 Mar;44(2):181-8. • J Pineal Res. 2007 Oct;43(3):225-31. • Am J Pathol. 2007 Feb;170(2):667-79.
– Mitochondria are especially sensitive
Valproic acid
• Is an anti-seizure medication • Patients on Valproic acid have been shown to have:
– Elevated homocysteine and decreased folate
• • • • • • • Prog Neuropsychopharmacol Biol Psychiatry. 2008 Apr 1;32(3):844-8 Epilepsy Res. 2006 Oct;71(2-3):229-32 Seizure. 2006 Mar;15(2):79-85 Eur J Paediatr Neurol. 2000;4(6):269-77 Epilepsy Res. 2000 Oct;41(3):253-7 Brain Dev. 2003 Mar;25(2):113-5. Acta Neurol. Scand. 69:226-231
Hcy
folate
– Alterations in the glutathione antioxidant enzymes
• J Basic Clin Physiol Pharmacol 2000;11(1):73-81 • Neuropediatrics 1998 Aug;29(4):195-20 • Br J Clin Pharmacol. 2004 Nov;58(5):542-7.
Valproic acid
• An animal study of valproic acid showed that within one hour of receiving a single injection of valproate:
– Significant reductions in hepatic SAM/SAH – Increase in hepatic GSSG (oxidized glutathione) – Decrease in plasma vitamin B-6
• Full text available at: http://jn.nutrition.org/cgi/reprint/132/9/2737
• A randomized, double-blind, placebo controlled trial of melatonin add-on therapy in epileptic children on valproate monotherapy: effect on glutathione peroxidase and glutathione reductase enzymes
– I recommend you read the full text avaialble at:
http://www.pubmedcentral.nih.gov/picrender.fcgi?artid=1884631&blobtype=pd
Valproic acid
• Gestational valproic acid exposure is used as an animal model for autism as well as being implicated in the development of autism in human fetuses exposed during critical time points during gestational development. – Proc Natl Acad Sci U S A. 2007 Aug 14;104(33):13501-6
– Full text available at:
http://www.pubmedcentral.nih.gov/picrender.fcgi?artid=1948920&blobtype=pdf
Big Question
• Can autism result from other environmental exposures or conditions which cause a similarity to the abnormal biochemistry induced by gestational valproic acid?
Abnormal transmethylation/transsulfuration metabolism and DNA hypomethylation among parents of children with autism.
Stratified group Control Mothers N (%) Case Mothers N (%) Odds ratio (95% CI)
SAH > 30 uMol/l
28 (14)
25 (54.3)
7.3 (3.6, 14.8)
SAM/SAH < 2.5
20 (10)
25 (54.3)
10.7 (5.1, 22.5)
tGSH/GSSG < 20 SAM/SAH < 2.5 and tGSH/GSSG < 20
22 (11)
30 (65.2)
15.2 (7.2, 32.1)
3 (1.5)
19 (41.3)
46.2 (12.8, 166.5)
Autism Dev Disord. 2008 Nov;38(10):1966-75
What are the repercussions of prenatal mercury exposure
Toxicol Appl Pharmacol. 2008 Feb 15;227(1):147-54
This study showed that prenatal mercury exposure altered the postnatal development of the brain glutathione (GSH) antioxidant system.
Key points from the study
• The alterations in the brain GSH system endured when cerebral mercury levels decreased nearly to basal levels. • Abnormal development of the GSH system occurred even when prenatal mercury exposure was so low that it resulted in brain mercury levels indistinguishable from those observed in controls.
Toxicol Appl Pharmacol. 2008 Feb 15;227(1):147-54
Can elevated gestational homocysteine work in a way similar to prenatal mercury exposure
Symptoms of magnesium deficiency
• • • • • • • • • • • • •
Poor sleep patterns, insomnia, restless sleep with frequent awakenings Increased sensitivity to noise Cognitive disorder, poor memory, confusion, disorientation Increase in migraines Increased muscular hyperexcitability, muscle twitching, spasms Tingling (may be described as zapping), numbness Heart arrthymias, palpitations, mitral valve prolapse, angina Anxiety, panic attacks Depression Irritability Asthma Difficulty learning PMS
Magnesium deficiency causes:
• • • • • • • Hypersensitivity of the NMDA receptor Increased histamine release (allergies) Decrease in cell mediated immunity Increased inflammatory cytokines
• TNF, IL-1, IL-6, substance P
Increased tendency for seizures Increased insulin resistance Increased oxidative stress
Magnesium, glutathione & oxidative stress
GSH/GSSG
Oxidative stress
Hypertension 1999;34:76-82
Parental Magnesium
• Magnesium Research 2006; 19(1): 53-62
– Lower RBC-Mg was found in both mothers and fathers of children with autism
• There was a statistically significant correlation of the mother’s Mg to that of her child with autism
– >
mothers
Consistent with altered GSH/GSSG in autistic children and their
• Behavioral improvement in the child with autism correlated with increases in RBC-Mg while on Mg supplements. Off supplement, behavior regressed as RBC-Mg fell to base-line.
• No difference in serum Mg was found
Testing for Mg deficiency
• Serum Mg levels do NOT reflect tissue levels.
• This is very well documented in the medical literature, yet this is the method most doctors use to measure magnesium status. – Am Coll Nutr. 2004 Dec;23(6):732S-7S
– Circulation 1995;92:2190-2197 (full text at: http://www.exatest.com/Research.htm)
•
RBC-Mg (Red Blood Cell Magnesium)
– Normal RBC-Mg does not rule out tissue deficiency
• • Journal of Chronic Fatigue Syndrome, vol 4(2) 1998 Nutrition. 1997 Apr;13(4):376-7
Testing for Mg deficiency
• Percentage retention of a Mg load in 24 hr urine test
– This test assumes normal kidney function. – Increased catecholamines cause increased Mg excretion. – What can increase catecholamines?
• Stress • Lack of sleep • Both of these factors are present in many parents.
Testing for Mg deficiency
• Sublingual buccal cell Mg
– Accurately reflects ion levels in muscle, cardiac tissue and bone
•
J Am Coll Nutr. 2004 Dec;23(6):732S-7S
– More information can be found at: www.exatest.com/physicians.htm
Side Note
Sublingual buccal Mg
7 out of 8 children with autism were found to have low levels of sublingual buccal Mg
Data collected with Dr. Paul Cutler (DAN doctor)
glycogen
Anaerobic Glycolysis
glucose Glucose 6-phosphate Fructose 6-phosphate Fructose 1,6-bisphosphate
Glucose 1-phosphate
Glycolysis occurs in the cytosol of the cell.
Dihydroxyacetone phosphate
Glyceraldehyde 3-phosphate
1,3-Bisphosphoglycerate Places where magnesium is necessary Lactate Oxaloacetate Malate 3-phosphoglycerate 2-phosphoglycerate Phosphoenolpyruvate Pyruvate Acetyl-CoA Citrate Isocitrate
The citric acid cycle occurs in the matrix of the mitochondria.
Citric acid cycle
Fumarate Succinate Succinyl-CoA 2-Oxoglutarate
Journal of Nutritional Medicine (1992) 3, 49-59 available at:
Recommended Reading
I highly recommend you all read:
– Review and Hypothesis: Might Patients with the Chronic Fatigue Syndrome have Latent Tetany of Magnesium Deficiency
•Journal of Chronic Fatigue Syndrome, Vol 4(2) 1998 Mildred Seelig, MD, MPH •The full text can be found on line at: www.mgwater.com/clmd.shtml – You will see many commonalities between what you read in this CFS paper and what is known about autism.
I wanted a quick fix!
But I knew I didn’t want to take drugs. So I took supplements.
I was unable to replete my magnesium.
I tried various forms, dosages and routes:
Pills & powders Baths & topical IV magnesium
Magnesium…where are you?
Consuming delicious greens
www.rawfamily.com
Great tasting recipes!
My magnesium, HVA and HIAA all normalized with greens, but not with supplements!
This made me question the appropriateness of spending more money on more supplements to improve my health.
= EDNP
(Junk Food )
High Calorie
-------------Increasing consumption of EDNP foods
Am J Clin Nutr 2000;72:926-36
------Increasing homocysteine
Serum concentrations of vitamins A, E, C, and folate are inversely related to consumption of high calorie, nutrient poor foods.
Am J Clin Nutr 2000;72:926-36
While you cannot have a genetic epidemic, dietary deficiencies can unmask a genetic predisposition to injury from an environmental exposure.
It’s all in the packaging!
Magnesium Spinach
Magnesium packaged with: Protein, calcium, zinc, sodium,selenium,vit C, vit B-6, phosphorus,manganese, thiamin,riboflavin,vit K, niacin, pantothenic acid, folate,choline, betaine, vit A, luetin,fiber,alpha tocopherol,zeaxanthin, gamma tocopherol and many other phytochemicals
Magnesium citrate
Folate was first isolated in 1941 from spinach. It is named after the Latin word folium, meaning leaf.
Plasma vitamin C, cholesterol and homocysteine are associated with grey matter volume determined by MRI in non-demented old people. Neurosci Lett. 2003 May 8;341(3):173-6.
Lower brain grey matter was associated with: Lower blood cell folate Lower plasma vitamin C Higher homocysteine Higher cholesterol
EDNP food consumption
(Junk Food )
A word about phytonutrients
• Food and brain function
– Food for the aging mind (Please, please, please read this!)
• www.ars.usda.gov/is/AR/archive/aug07/aging0807.htm
• Reversing the deleterious effects of aging on neuronal communication and behavior: beneficial properties of fruit polyphenolic compounds.
• Am J Clin Nutr 2005 Jan;81(1 Suppl):313S-316S • full text available at http://www.ajcn.org/cgi/reprint/81/1/313S
• Molecular targets of dietary agents for prevention and therapy of cancer
• Biochemical Pharmacology 71(2006) 1397-1421
Confusion about what to eat?
Watch 90 min video of Colin Campbell at: http://www.youtube.com/watch?v=4XzDOGZKI68&feature=related
Nutrient Density
• A nutrient density standard for vegetables and fruits: nutrients per calorie and nutrients per unit cost.
J Am Diet Assoc. 2005 Dec;105(12):1881-7.
– “Energy density and nutrient density score were negatively correlated, confirming the widely accepted notion that energy-dense foods tend to be nutrient-poor.” – “… fruits and vegetables had the highest nutrient density score because they were nutrient-rich in relation to their low energy content.”
FIGURE 4. Relation between energy density and the naturally nutrient rich score for vegetables and fruit
Drewnowski, A. Am J Clin Nutr 2005;82:721-732
Copyright ©2005 The American Society for Nutrition
Used with permission
Health Equation
• Eat foods high in nutrients and low in calories. Health = Nutrient/Calories
– – Dr. Joel Furhman Video: The Greatest Diet on Earth
• www.DrFuhrman.com
•
•
Junk Food defined
– low nutrients with high calories
Gr eens
• Greens have the highest nutrient per calorie ratio of all fruits and vegetables. – www.peertrainer.com/diet/nutrient_density.html Greens are a good source of magnesium, folic acid and vitamin K. “Vitamin K intake may be a marker of a healthy diet because it is found mainly in green, leafy vegetables”. Only 27% of people in the United States consume the average daily goal for vit K.
– Food consumption survey analysis from the Human Nutrition Research Center, part of the Agricultural Research Service. Agricultural Research/August 2007
• •
• http://www.ars.usda.gov/is/AR/archive/aug07/vitamin0807.htm
ORAC Values
• ORAC: Oxygen Radical Absorbance Capacity • Method for measuring antioxidant capacity of foods • ORAC database for 277 food items at:
– http://www.ars.usda.gov/Services/docs.htm?docid=15866
Food description
Apple juice,unsweetened without added ascorbic acid Apple, Red Delicious,raw, without skin Apple, Red Delicious,raw with skin Applesauce,unsweetened, without added ascorbic acid Artichoke (globe or French) raw Artichoke (Ocean Mist) boiled Blueberries, raw
Total-ORAC
408 2936
Calories
47 Kcal 48 Kcal
4275
52 Kcal
1965
43 Kcal
6552 9416
47 Kcal
53 Kcal
6552
57 Kcal
Data derived from: http://www.ars.usda.gov/Services/docs.htm?docid=15866
As we age, our bodies produce and accumulate a family of molecules called Advanced Glycation End-products.
Increasi ng AG Es
Gl ucos Protei n e Advanced Glycation End-product (AGE) Gl ucos e Li pi d s
Advanced Lipoxidation End-product (ALE)
Glycotoxins
Facts about AGEs
• AGEs cause proteins or lipids to be structurally altered in such a way as to make the protein or lipid dysfunctional.
– Hand analogy with cross-linking
• AGEs are proinflammatory and promote oxidative stress. • AGEs are associated with pathological conditions.
• Diabetes, renal failure, artherosclerosis, Alzheimer’s, rheumatoid arthritis, cancer,cataracts, allergic and autoimmune diseases
Food derived AGEs
• AGEs derived from food are absorbed and contribute to serum AGE levels in both animal and human studies. Ann N Y Acad Sci. 2005 Jun;1043:452-60
Increased AGEs in serum
QuickTimeª and a TIFF (Uncompressed) decompressor are needed to see this picture.
Food derived AGEs
• Once thought to only be problematic in persons with diabetes or renal failure, dietary AGEs are now being shown to cause inflammation and contribute to inflammatory and degenerative processes even in healthy individuals.
– – Diet-derived advanced glycation end products are major contributors to the body's AGE pool and induce inflammation in healthy subjects. Ann N Y Acad Sci. 2005 Jun;1043:461-6. Circulating glycotoxins and dietary advanced glycation endproducts: two links to inflammatory response, oxidative stress, and aging. J Gerontol A Biol Sci Med Sci. 2007 Apr;62(4):427-33.
Advanced glycoxidation end products in commonly consumed foods. J Am Diet Assoc. 2004
Aug;104(8):1287-91
AGE production in foods is temperature and moisture dependent.
•AGEs in a 90 gram serving of chicken breast
–9,000 kU oven frying –6,700 kU deep frying –5,250 kU broiling –4,300 kU roasting –1,000 kU boiling
Boiling, poaching, stewing vs. dry heat
So up …. .a be tter ch oic e!
Foods highest in fat contain the most AGEs, followed by meats, processed carbohydrates and then lowest in AGE content are vegetables and fruits. 1 lb = 454 grams But ter 265 kU /g Chi cken breast broi led 58 kU/ g Kel logs Rice Kri spi es 18kU /g Green beans 0. 18 kU /g Banana 0. 01 (not to scale) /g kU
J Am Diet Assoc. 2004 Aug;104(8):1287-91
Advanced glycoxidation end products in commonly consumed foods. J Am Diet Assoc. 2004
Aug;104(8):1287-91
AGEs in whole human milk 0.05 kU/ml
AGEs in Enfamil Infant Formula 4.86 kU/ml
“…since AGEs are known immune cell modulators, the introduction of infant diets, rich in AGE antigens, may account for the rise in childhood autoimmune diseases such as Type 1 Diabetes.”
Side Note
Breast-feeding reduces risk of:
Acute otitis media Non-specific gastroenteritis Lower respiratory tract infections Atopic dermatitis Asthma (young children) Obesity Type 1 and 2 diabetes Childhood leukemia Sudden infant death syndrome Necrotizing enterocolitis
Evid Rep Technol Assess . 2006 Apr;(134):1-161.
Hiding AGEs
• Unexpected places for finding food AGEs
– Soy sauce 8,700 AGE, units /15 ml
• (3 teaspoons)
– Classic Coca-Cola 8,500 AGE, units/cup – Diet Coke (soda) 9,500 AGE, units/cup
Proc Natl Acad Sci USA 1997 June 10; 94(12): 6474-6479
Regular AGE Diet
Same foods different temperatures used during preparation.
Low AGE Diet
25% decrease in GSH/GSSG
Basel ine compared to 24 months 40% increase in GSH/GSSG
45% decrease in 8-isoprostanes Increased AGER1 expression Unaltered RAGE expression Extended lifespan
(equal to mice on 40% calorie restriction)
20% increase in 8-isoprostanes Decreased AGER1 expression Increased RAGE expression Regular lifespan
Am J Pathol. 2007 Jun;170(6):1893-902. Full text available at: www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=17525257
Side note: AGEs in autism
• Proteomic studies identified a single nucleotide polymorphism in glyoxalase I as autism susceptibility factor. Am J Med Genet A. 2004 Nov
15;131(1):11-7.
– Analyses of autopsied autism brains show AGE accumulation.
• These were not brains from aged persons.
– 38% decrease in Glo1 enzyme activity in autism brains versus controls
• Results in accumulation of brain AGEs
•
Alterations of circulating endogenous secretory RAGE and S100A9 levels indicating dysfunction of the AGE-RAGE axis in autism. Neurosci
Lett. 2006 Dec 27;410(3):169-73.
– If an AGE binds to a RAGE receptor it initiates oxidative stress and inflammation. – esRAGE receptors can act as decoys to prevent AGEs from binding to the RAGE receptors. – Peripheral esRAGE levels were significantly reduced in autism.
Dietary AGEs
Self-perpetuating cycle of chronic inflammation
Adapted from a slide by Dr. Jill James (used with permission)
Heavy Metals Genetics Viral infection Inflammation GSH/GSSG
Acute Inflammation (Resolution)
PPAR-RXR dimer (downregulates transcription) TNFα IL-6 IL-1 PGJ(2) GSH/GSSG OFF-SWITCH iNOS
low
ROS
Free Radicals Transcription Cytokines factors
NFkB AP-1
NO
Nitric oxide
high
GSH/GSSG
Th1 Th2
Chronic Inflammation
Di etary AGEs Environmental insults Infection Genetic polymorphisms Dietary deficiencies
Oxi dati v e stres s
Ant ioxidan ts
Advanced glycation end products and RAGE: a common thread in aging, diabetes, neurodegeneration, and inflammation.
Glycobiology. 2005 Jul;15(7):16R-28R.
Full text available at:
http://glycob.oxfordjournals.org/cgi/content/full/15/7/16R
Break Time (5 Min)
When we return we will be discussing the “how” mechanisms of the biochemical pathways involving folate and glutathione.
The Folate Cycle
Pyrimidine synthesis Uracil Thymine
Folate sufficiency
Purine synthesis Adenine & Guanine
Folate deficiency
Methionine 10-formyl-THF 5,10-methylene THF MTHFR DHF DHFR THF B12
Methyl THF
MS
homocysteine
Double Strand Breaks
• DNA deletions • DNA translocations
The Folate Cycle
Purine synthesis Adenine & Guanine
Folate deficiency
Pyrimidine synthesis Uracil Thymine
Folate sufficiency
Methionine 10-formyl-THF DHF DHFR 5,10-methylene THF MTHFR THF B12
Methyl THF
homocysteine
MS
Methionine synthase
5,10-methylene THF MTHFR
Methyl THF
CH3THF
Methionine THF B12 MS homocysteine
Step 1:
CH3 THF
CH3 Cobalamin (B12) (Methylcobalamin)
Step 2:
CH3 Cobalamin (B12)
CH3 homocysteine (Methionine)
methionine Folate cycle SAM THF
B12
Methylation Cycle
MS
SAH homocysteine
(Methyl THF)
CH3-THF
MS : methionine synthase SAM : Sadenosylmethionine SAH : Sadenosylhomocysteine
Notes on methylation
CH3 is called a methyl group SAM is called a universal methyl donor A family of enzymes called methyl transferases serve as the working force to transfer methyl groups from SAM onto other molecules
Methyl transferase taxi analogy
Used in myelin basic protein Convert serotonin to melatonin Convert norepinephrine to epinephrine
Catecholamine
detoxification via COMT
Methylation in the liver is a detoxification pathway for many chemicals.
Gene expression
Implications for cancer
Viral silencing
Decreasing SAM/SAH ratio
Inhibits methyl transferase activity
methionine Folate cycle SAM THF
B12
Methylation Cycle
MS
SAH homocysteine
B6
CH3-THF
(Methyl THF)
cystathionine
B6
cysteine glutathione
Transsulfuration pathway
H 2O H 2O 2
Hydrogen Peroxide
Hydrogen Peroxide
+
Glutathione
Glutathione Utilization
cysteine
SOD GST
Detoxification of drugs and chemicals
H2O2
GPx
GSH
GR
H2O
GSSG
GSH: glutathione GSSG: oxidized glutathione GR: glutathione reductase GPx: glutathione peroxidase
GST: glutathione transferase SOD: superoxide dismutase H2O2: hydrogen peroxide
Notes on Glutathione
Is an antioxidant made in your body Converts hydrogen peroxide to harmless water Environmental chemicals, heavy metals, dietary deficiencies and medical drugs can deplete glutathione Glutathione has antiviral properties Lower levels found in children with autism compared to control children Protects DNA and mitochondria
What happens if hydrogen peroxide production exceeds glutathione production?
Hydrogen Peroxide glutathione Hydrogen Peroxide Hydrogen Peroxide Hydrogen Peroxide glutathione Hydrogen Peroxide
glutathione
Hydrogen Peroxide
Art by: Christy Domino
Metals Pesticides Chemicals Infections Poor diet
O2- (super oxide) OH (hydroxyl) NO (Nitric Oxide) OONO- (peroxynitrite) H2O2 (hydrogen peroxide) Damage to: DNA, proteins, lipids
oxidants
antioxidants
No oxidative stress
Free radical production in balance with antioxidants
Environmental insults Infection Genetic polymorphisms Poor diet
Oxi da tive str ess
Ant ioxidants
Environmental insults can severely tip the scale in those with increased vulnerability
• Folic acid impacts cancer risk in the following ways:
Cancer
– DNA methylation (gene expression) – DNA synthesis
• Uracil and DNA repair
– Detoxification of chemicals and control of oxidative stress
• Methylation of chemicals • Glutathione (GSH)
– GST helps inactivate chemical carcinogens into less toxic or inactive metabolites
methionine THF
B12 MS
SAM SAH H homocysteine H cystathionine cysteine L GSH
GPx GST
CH3-THF
(Methyl THF)
High and low values of ASD parents compared to controls.
SOD
H2O2
H2O
H GSSG
GR
thimerosal
Does dose make the poison
GSH
GSH
GSH
GSH
GS H
thimerosal
GS
H
GSH
GSH
Glutathione (GSH) protects against thimerosal induced apoptosis (cell death)
Genes Immun 2002 Aug;3(5):270-8
GSH
GSH
Cell Death
thimerosal
GSH
SAME DOSE of thimerosal as in previous slide!
A person with lower levels of GSH
GSH
QuickTimeª and a decompressor are needed to see this picture.
Low glutathione levels can make people more sensitive to DNA damage from a variety of mutagenic environmental exposures.
A s pecia l Th ank Yo u to Dr. J ill James Dr. P aul Cutle r My fr iend Pa ula Teri A rr anga Ray J anak And to yo u…t he pare nts !
Laurette Janak Autism One May 2009 Part 1 of 2 Laurette.janak@verizon.net
Overview
Part One: the “What” What has been found? What it means? What to do about it?
Part Two: the “How” How the biochemical pathways for folate and glutathione function.
Metabolic Findings in Parents of Children with Autism
• 86 autism parents differ from 200 controls in the following:
– Higher homocysteine (Hcy) – Higher SAH (S-adenosylhomocysteine) – Lower GSH (glutathione) – Increased GSSG (oxidized glutathione)
J Autism Dev Disord. 2008 Nov;38(10):1966-75
Mothers of Autistic Children
J Autism Dev Disord. 2008 Nov;38(10):1966-75
46 Case Moms Mean Homocysteine Mean SAH Mean total GSH Mean GSSG 9.8 * 33 * 5.1 * .32 *
200 Control Moms 7.4 23 7.3 .24
* p < 0.001 Case Versus Control Mothers
Risk vs. Causation
Elevated Homocysteine
• What does elevated homocysteine mean? • It is a marker for a folic acid and/or vitamin B12 deficiency. (B-6) • What are the health implications of elevated homocysteine?
Homocysteine, folate, methylation and monoamine metabolism in depression
–J Neurol Neurosurg Pyschiatry 2000;69(2):228-32
52% of the depressed persons in this study had elevated homocysteine
Full text available at: http://www.pubmedcentral.nih.gov/picrender.fcgi?artid=1737050&blobtype=pdf
Depression
“Relatives of autistic individuals have high rates of major depression and social phobia that are not associated with the broad autism phenotype and cannot be explained by the increased stress associated with raising an autistic child.”
Am J Psychiatry 1999 Apr;156(4):557-63
Full text available at: http://ajp.psychiatryonline.org/cgi/reprint/156/4/557
Depression
• “Folic acid is a simple method of greatly improving the antidepressant action of fluoxetine…. Folic acid should be given in doses sufficient to decrease plasma homocysteine.”
– Enhancement of the antidepressant action of fluoxetine by folic acid: a randomized, placebo controlled trial
• J Affect Disord 2000 Nov;60(2):121-30
• What if the doctor does NOT screen you for homocysteine before putting you on a SSRI?
Depression Linked to Heart Disease
Depression has been shown to be an independent risk factor for coronary artery disease and vice versa.
–Arch Gen Psyschiatry 2004 Apr;61(4):369-76. –Eur Heart J. 2004 Jan;25(1):3-9. –Prostaglandins Leukot Essent Fatty Acids 2004 Apr;70(4):56,349
Cardiovascular Disease
• Elevated homocysteine is an independent risk factor for vascular disease
– Hyperhomocysteinemia: an independent risk factor for vascular disease
» N Engl J Med 1991 Apr 25;324(17):1149-55
– Plasma total homocysteine and cardiovascular and noncardiovascular mortality: the Hordaland Homocysteine Study.
» Am J Clin Nutr 2001 Jul;74(1):130-6
•
Folate and homocysteine metabolism: therapeutic targets in cardiovascular and neurodegenerative disorders.
» Curr Med Chem 2003 Oct;10(19):1923-9
Alzheimer’s Disease
• Elevated homocysteine is an independent risk factor for Alzheimer's disease.
– Neuroscience 2007 Mar 30;14593):942-54 – N Engl J Med. 2002 Feb 14;346(7):476-83. – Neuroreport. 2003 Jul 18;14(10):1391-4.
Parkinson’s Disease
• Disturbances in folate metabolism have been implicated as a risk factor for Parkinson’s.
– – – – – – – – Int J Mol Med. 2004 Mar;13(3):343-53 J Neurol Sci. 2003 Mar 15;207(1-2):19-23. Neurology. 2003 Feb 25;60(4):690-5. Trends Neurosci. 2003 Mar;26(3):137-46. Nutr Rev. 2002 Dec;60(12):410-3. J Neural Transm. 2002 Dec;109(12):1445-52. Int J Geriatr Psychiatry. 2002 Sep;17(9):859-64. J Neurochem. 2002 Jan;80(1):101-10.
Michael J. Fox
Schizophrenia and Homocysteine
• Elevated third trimester homocysteine levels have been associated with increased risk for schizophrenia in the offspring.
– Full text available at: http://archpsyc.ama-assn.org/cgi/reprint/64/1/31
• An increased occurrence of schizophrenia has been reported in parents of children with autism when compared to control parents.
– Pediatrics. 2008 May;121(5):e1357-62
• In a randomized, double-blind, placebo-controlled, crossover designed study, 42 schizophrenic patients with elevated homocysteine were treated with folic acid, B-12, and B-6
– The treatment resulted in significant clinical improvements
• Biol Psychiatry. 2006 Aug 1;60(3):265-9
Cancer
“Most families report cancer in the parents or grandparents, the most common being colon cancer.”
–Autism and vaccinations by Mary Megson, MD. •Wise Traditions Fall 2000
Increased mortality from cancer in individuals with autism.
–Causes of death in autism. •J Autism Dev Disord. 2001 Dec;31(6):569-76
Cancer
• Epidemiology studies have shown an association of reduced folate intake with an increased occurrence of certain cancers.
Cancer
Folic acid used as a chemopreventative agent has shown that results depend on the timing of the folic acid supplementation. For example:
Folic acid supplementation prior to any neoplastic lesions was shown to be protective against colon cancer. Folic acid supplementation once neoplastic lesions are present has been shown to fuel colon cancer growth.
Fibromyalgia Myth
• “Fibromyalgia is essentially a benign disease. It is not a degenerative or deforming condition, nor does it result in life-threatening complications.”
– (Patient information: Fibromyalgia from the Cleveland Clinic 6/22/06 Author Don L Goldenberg, MD published in www.uptodate.com)
FM patients have less total gray matter in their brains than controls. J Neurosci 2007 Apr 11;27(15):4004-7
–The longer they have had FM, the greater the gray matter loss
–Each year of FM = 9.5 times the gray matter loss as that seen in normal aging
Lower brain grey matter has been found to be associated with: Lower blood cell folate Higher homocysteine
Neurosci Lett. 2003 May 8;341(3):173-6.
Fibromyalgia Myth (It’s all in your head)
Fibromyalgia Myth
(It’s all in your head)
• “the results do confirm that there are real metabolic changes in painful muscles in PF patients.”
– (PF = primary fibromyalgia) – This study was done on muscle biopsies
• Arthritis and Rheumatism, Vol 29, No. 7 (July 1986)
•
Use of P-31 Magnetic Resonance Spectroscopy to Detect Metabolic Abnormalities in Muscles of Patients with Fibromyalgia.
– This study reiterates the findings of the 1986 study using MRS technology instead of muscle biopsy. – Findings: “impaired mitochondrial function”
•
Arthritis & Rheumatism vol 41, No3, March 1998 pp 406-413
Mitochondria produce energy (ATP)
• Muscle studies in FM have shown mitochondrial damage.
– – – – Joint Bone Spine. 2006 May;73(3):239-42. Ann Rheum Dis. 2004 Mar;63(3):245-51. Curr Rheumatol Rep. 2000 Apr;2(2):131-40. Z Rheumatol. 1998;57 Suppl 2:47-51.
Cristae - the site of the electron transport chain Matrix- the site of the citric acid cycle
Myth
(It’s all in your head)
Using either muscle biopsy or 31P magnetic resonance spectroscopy the following are found with respect to FM or CFS:
These values are increased:
Inorganic phosphate (Pi) Phosphodiesters (PDE) Pyruvate Lactate Glycolysis iNOS Increased sensitivity to NO induced suppression of oxidative phosphorylation. DNA fragmentation Lipid accumulation Lipofuscin Oxidative damage to lipids in muscle Activity of the antioxidant enzymes catalase, glutathione peroxidase and transferase
These values are decreased:
Phosphocreatine (PCr) ATP Phosphorylation potential (PP) Oxidative capacity (Vmax) Lactate dehydrogenase (LDH) Reduced nutritive blood flow during aerobic exercise Number of mitochondria. Altered morphology of mitochondria Intramuscular collagen Hydroxyproline (in muscle) Lower total concentration of the major amino acids of collagen than in controls (in muscle) Acetyl l-carnitine
Data I collected from multiple studies cited in Pubmed
Homocysteine and Mitochondria
• Correlation of plasma homocysteine and mitochondrial DNA content in peripheral blood in healthy women Atherosclerosis. 2001 Oct;158(2):399-405.
– A physiologic range of total plasma homocysteine (tHcy) was shown to have the following significant relationship:.
tHcy
mtDNA content
Chronic Fatigue Syndrome (CFS) and Homocysteine Is there a connection?
• Elevated homocysteine was found in cerebrospinal fluid of women who fulfilled the criteria for both FM and CFS.
– There was a significant correlation between the CSF-Hcy levels and fatigability. – Scand J Rheumatol 1997;26(4):301-7
•
Plasma homocysteine vs cerebrospinal fluid homocysteine
Parental Oxidative Stress
• Parents of children with autism were found to have:
– Significantly decreased levels of GSH (glutathione) – Significantly increased levels of GSSG (oxidized glutathione) – Significantly decreased ratio of GSH/GSSG
• J Autism Dev Disord. 2008 Nov;38(10):1966-75
• This means the parents are under oxidative stress.
Oxidative Stress and Psychiatric Disorders
• A meta-analysis indicates an association of oxidative stress in the majority of DSM-IV psychiatric disorders including: autism, Rett’s, ADHD, schizophrenia, anxiety, and mood disorders. Full text available at:
http://www.pubmedcentral.nih.gov/picrender.fcgi?artid=2330073&blobtype=pdf
• “…all these psychiatric disorders might benefit from a change to a whole-food plant-based diet.”
Oxidat ive Stress in Chr on ic F at igue Erythrocyte oxidative damage in chronic fatigue syndrome. Synd rome Arch Med Res. 2007 Jan;38(1):948.
Chronic fatigue syndrome is accompanied by an IgMrelated immune response directed against neopitopes formed by oxidative or nitrosative damage to lip ids and proteins. Neuro Endocrinol Lett. 2006 Oct;27(5):61521. Oxidative stress levels are raised in chronic fatigue syndrome and are associated with clin ical symptoms. Free Radic Biol Med. 2005 Sep 1;39(5):5849. Chronic fatigue syndrome: assessment of increased oxidative stress and altered muscle excitabil ity in response to incremental exercise. J Intern Med. 2005 Mar;257(3):299310. Elevated levels of protein carbonyls in sera of chronic fatigue syndrome patients. Mol Cell Biochem. 2003 Jun;248(12):935. Effect of natural and synthetic antioxidants in a mouse model of chronic fatigue syndrome. J Med Food. 2002 Winter;5(4):21120. Relationship between musculoskeletal symptoms and blood markers of oxidative stress in patients with chronic fatigue syndrome. Neurosci Lett. 2003 Jan 2;335(3):1514. Elevated nitric oxide/peroxynitrite mechanism for the common etiology of multiple chemical sensitivity, chronic fatigue syndrome, and posttraumatic stress disorder. Ann N Y Acad Sci. 2001 Mar;933:3239 Chronic fatigue syndrome: oxidative stress and dietary modifications. Altern Med Rev. 2001 Oct;6(5):4509 Specific oxidative alterations in vastus lateralis muscle of patients with the diagnosis of chronic fatigue syndrome. Free Radic Biol Med. 2000 Dec 15;29(12):12529. Blood parameters indicative of oxidative stress are associated with symptom expression in chronic fatigue syndrome. Redox Rep. 2000;5(1):3541.
Chronic Fatigue Syndrome
• “CFS patients have a lipid profile and oxidant biology that is consistent with cardiovascular risk….Supplementation with specific antioxidant medications might help to ameliorate symptoms and any potential cardiovascular complications of the illness.”
– Free Radical Biology & Medicine 39 (2005) 584-589
Oxidative Stress in Fibromyalgia
Total antioxidant capacity and the severity of the pain i n patients with fibromyalgia. Redox Rep. 2006;11(3):1315. Current concepts in the pathoph ysiology of fibromyalgia: the potential role of oxidative stress and nitric oxide. Rheumatol Int. 2006 May;26(7):58597. Antioxidant status, lipid peroxidation and n itric oxide in fibromyalgia: etiologic and therapeutic concerns. Rheumatol Int. 2006 May;26(7):598603. Free radicals and antioxidants in primary fibromyalgia: an oxidative stress disorder? Rheumatol Int. 2005 Apr;25(3):18890. Are advanced glycation endproductmodified proteins of pathogenetic importance in fibromyalgia? Rheumatology (Oxford). 2002 Oct;41(10):11637.
•
Oxidative stress is implicated in endometriosis.
– – – – J Soc Gynecol Investig. 2006 Sep;13(6):390-8 Reprod Biomed Online. 2006 Jul;13(1):126-34 Hum Reprod. 2005 Jul;20(7):2014-20. Fertil Steril. 2004 Oct;82 Suppl 3:1019-22.
•
High rates of autoimmune and endocrine disorders, fibromyalgia, chronic fatigue syndrome and atopic diseases among women with endometriosis: a survey analysis Hum Reprod. 2002 Oct;17(10):2715-24.
• “CONCLUSIONS: Hypothyroidism, fibromyalgia, chronic fatigue syndrome, autoimmune diseases, allergies and asthma are all significantly more common in women with endometriosis than in women in the general USA population.” – Full text online at: http://humrep.oxfordjournals.org/cgi/content/full/17/10/2715
• Mitochondrial dysfunction and molecular pathways of disease Exp Mol Pathol. 2007 Jan 17
– “A wide range of seemingly unrelated disorders, such as schizophrenia, bipolar disease, dementia, Alzheimer's disease, epilepsy, migraine headaches, strokes, neuropathic pain, Parkinson's disease, ataxia, transient ischemic attack, cardiomyopathy, coronary artery disease, chronic fatigue syndrome, fibromyalgia, retinitis pigmentosa, diabetes, hepatitis C, and primary biliary cirrhosis, have underlying pathophysiological mechanisms in common, namely reactive oxygen species (ROS) production, the accumulation of mitochondrial DNA (mtDNA) damage, resulting in mitochondrial dysfunction. Antioxidant therapies hold promise for improving mitochondrial performance.”
Homocysteine and Brain Development
• An animal model of maternal elevated homocysteine produced offspring with:
– Altered fetal brain development – Reduced learning abilities – Increased lipid peroxidation in the brain
• Indicates oxidative stress
– Increased brain DNA fragmentation – Abnormalities mitigated by using melatonin
• J Pineal Res. 2008 Mar;44(2):181-8. • J Pineal Res. 2007 Oct;43(3):225-31. • Am J Pathol. 2007 Feb;170(2):667-79.
– Mitochondria are especially sensitive
Valproic acid
• Is an anti-seizure medication • Patients on Valproic acid have been shown to have:
– Elevated homocysteine and decreased folate
• • • • • • • Prog Neuropsychopharmacol Biol Psychiatry. 2008 Apr 1;32(3):844-8 Epilepsy Res. 2006 Oct;71(2-3):229-32 Seizure. 2006 Mar;15(2):79-85 Eur J Paediatr Neurol. 2000;4(6):269-77 Epilepsy Res. 2000 Oct;41(3):253-7 Brain Dev. 2003 Mar;25(2):113-5. Acta Neurol. Scand. 69:226-231
Hcy
folate
– Alterations in the glutathione antioxidant enzymes
• J Basic Clin Physiol Pharmacol 2000;11(1):73-81 • Neuropediatrics 1998 Aug;29(4):195-20 • Br J Clin Pharmacol. 2004 Nov;58(5):542-7.
Valproic acid
• An animal study of valproic acid showed that within one hour of receiving a single injection of valproate:
– Significant reductions in hepatic SAM/SAH – Increase in hepatic GSSG (oxidized glutathione) – Decrease in plasma vitamin B-6
• Full text available at: http://jn.nutrition.org/cgi/reprint/132/9/2737
• A randomized, double-blind, placebo controlled trial of melatonin add-on therapy in epileptic children on valproate monotherapy: effect on glutathione peroxidase and glutathione reductase enzymes
– I recommend you read the full text avaialble at:
http://www.pubmedcentral.nih.gov/picrender.fcgi?artid=1884631&blobtype=pd
Valproic acid
• Gestational valproic acid exposure is used as an animal model for autism as well as being implicated in the development of autism in human fetuses exposed during critical time points during gestational development. – Proc Natl Acad Sci U S A. 2007 Aug 14;104(33):13501-6
– Full text available at:
http://www.pubmedcentral.nih.gov/picrender.fcgi?artid=1948920&blobtype=pdf
Big Question
• Can autism result from other environmental exposures or conditions which cause a similarity to the abnormal biochemistry induced by gestational valproic acid?
Abnormal transmethylation/transsulfuration metabolism and DNA hypomethylation among parents of children with autism.
Stratified group Control Mothers N (%) Case Mothers N (%) Odds ratio (95% CI)
SAH > 30 uMol/l
28 (14)
25 (54.3)
7.3 (3.6, 14.8)
SAM/SAH < 2.5
20 (10)
25 (54.3)
10.7 (5.1, 22.5)
tGSH/GSSG < 20 SAM/SAH < 2.5 and tGSH/GSSG < 20
22 (11)
30 (65.2)
15.2 (7.2, 32.1)
3 (1.5)
19 (41.3)
46.2 (12.8, 166.5)
Autism Dev Disord. 2008 Nov;38(10):1966-75
What are the repercussions of prenatal mercury exposure
Toxicol Appl Pharmacol. 2008 Feb 15;227(1):147-54
This study showed that prenatal mercury exposure altered the postnatal development of the brain glutathione (GSH) antioxidant system.
Key points from the study
• The alterations in the brain GSH system endured when cerebral mercury levels decreased nearly to basal levels. • Abnormal development of the GSH system occurred even when prenatal mercury exposure was so low that it resulted in brain mercury levels indistinguishable from those observed in controls.
Toxicol Appl Pharmacol. 2008 Feb 15;227(1):147-54
Can elevated gestational homocysteine work in a way similar to prenatal mercury exposure
Symptoms of magnesium deficiency
• • • • • • • • • • • • •
Poor sleep patterns, insomnia, restless sleep with frequent awakenings Increased sensitivity to noise Cognitive disorder, poor memory, confusion, disorientation Increase in migraines Increased muscular hyperexcitability, muscle twitching, spasms Tingling (may be described as zapping), numbness Heart arrthymias, palpitations, mitral valve prolapse, angina Anxiety, panic attacks Depression Irritability Asthma Difficulty learning PMS
Magnesium deficiency causes:
• • • • • • • Hypersensitivity of the NMDA receptor Increased histamine release (allergies) Decrease in cell mediated immunity Increased inflammatory cytokines
• TNF, IL-1, IL-6, substance P
Increased tendency for seizures Increased insulin resistance Increased oxidative stress
Magnesium, glutathione & oxidative stress
GSH/GSSG
Oxidative stress
Hypertension 1999;34:76-82
Parental Magnesium
• Magnesium Research 2006; 19(1): 53-62
– Lower RBC-Mg was found in both mothers and fathers of children with autism
• There was a statistically significant correlation of the mother’s Mg to that of her child with autism
– >
mothers
Consistent with altered GSH/GSSG in autistic children and their
• Behavioral improvement in the child with autism correlated with increases in RBC-Mg while on Mg supplements. Off supplement, behavior regressed as RBC-Mg fell to base-line.
• No difference in serum Mg was found
Testing for Mg deficiency
• Serum Mg levels do NOT reflect tissue levels.
• This is very well documented in the medical literature, yet this is the method most doctors use to measure magnesium status. – Am Coll Nutr. 2004 Dec;23(6):732S-7S
– Circulation 1995;92:2190-2197 (full text at: http://www.exatest.com/Research.htm)
•
RBC-Mg (Red Blood Cell Magnesium)
– Normal RBC-Mg does not rule out tissue deficiency
• • Journal of Chronic Fatigue Syndrome, vol 4(2) 1998 Nutrition. 1997 Apr;13(4):376-7
Testing for Mg deficiency
• Percentage retention of a Mg load in 24 hr urine test
– This test assumes normal kidney function. – Increased catecholamines cause increased Mg excretion. – What can increase catecholamines?
• Stress • Lack of sleep • Both of these factors are present in many parents.
Testing for Mg deficiency
• Sublingual buccal cell Mg
– Accurately reflects ion levels in muscle, cardiac tissue and bone
•
J Am Coll Nutr. 2004 Dec;23(6):732S-7S
– More information can be found at: www.exatest.com/physicians.htm
Side Note
Sublingual buccal Mg
7 out of 8 children with autism were found to have low levels of sublingual buccal Mg
Data collected with Dr. Paul Cutler (DAN doctor)
glycogen
Anaerobic Glycolysis
glucose Glucose 6-phosphate Fructose 6-phosphate Fructose 1,6-bisphosphate
Glucose 1-phosphate
Glycolysis occurs in the cytosol of the cell.
Dihydroxyacetone phosphate
Glyceraldehyde 3-phosphate
1,3-Bisphosphoglycerate Places where magnesium is necessary Lactate Oxaloacetate Malate 3-phosphoglycerate 2-phosphoglycerate Phosphoenolpyruvate Pyruvate Acetyl-CoA Citrate Isocitrate
The citric acid cycle occurs in the matrix of the mitochondria.
Citric acid cycle
Fumarate Succinate Succinyl-CoA 2-Oxoglutarate
Journal of Nutritional Medicine (1992) 3, 49-59 available at:
Recommended Reading
I highly recommend you all read:
– Review and Hypothesis: Might Patients with the Chronic Fatigue Syndrome have Latent Tetany of Magnesium Deficiency
•Journal of Chronic Fatigue Syndrome, Vol 4(2) 1998 Mildred Seelig, MD, MPH •The full text can be found on line at: www.mgwater.com/clmd.shtml – You will see many commonalities between what you read in this CFS paper and what is known about autism.
I wanted a quick fix!
But I knew I didn’t want to take drugs. So I took supplements.
I was unable to replete my magnesium.
I tried various forms, dosages and routes:
Pills & powders Baths & topical IV magnesium
Magnesium…where are you?
Consuming delicious greens
www.rawfamily.com
Great tasting recipes!
My magnesium, HVA and HIAA all normalized with greens, but not with supplements!
This made me question the appropriateness of spending more money on more supplements to improve my health.
= EDNP
(Junk Food )
High Calorie
-------------Increasing consumption of EDNP foods
Am J Clin Nutr 2000;72:926-36
------Increasing homocysteine
Serum concentrations of vitamins A, E, C, and folate are inversely related to consumption of high calorie, nutrient poor foods.
Am J Clin Nutr 2000;72:926-36
While you cannot have a genetic epidemic, dietary deficiencies can unmask a genetic predisposition to injury from an environmental exposure.
It’s all in the packaging!
Magnesium Spinach
Magnesium packaged with: Protein, calcium, zinc, sodium,selenium,vit C, vit B-6, phosphorus,manganese, thiamin,riboflavin,vit K, niacin, pantothenic acid, folate,choline, betaine, vit A, luetin,fiber,alpha tocopherol,zeaxanthin, gamma tocopherol and many other phytochemicals
Magnesium citrate
Folate was first isolated in 1941 from spinach. It is named after the Latin word folium, meaning leaf.
Plasma vitamin C, cholesterol and homocysteine are associated with grey matter volume determined by MRI in non-demented old people. Neurosci Lett. 2003 May 8;341(3):173-6.
Lower brain grey matter was associated with: Lower blood cell folate Lower plasma vitamin C Higher homocysteine Higher cholesterol
EDNP food consumption
(Junk Food )
A word about phytonutrients
• Food and brain function
– Food for the aging mind (Please, please, please read this!)
• www.ars.usda.gov/is/AR/archive/aug07/aging0807.htm
• Reversing the deleterious effects of aging on neuronal communication and behavior: beneficial properties of fruit polyphenolic compounds.
• Am J Clin Nutr 2005 Jan;81(1 Suppl):313S-316S • full text available at http://www.ajcn.org/cgi/reprint/81/1/313S
• Molecular targets of dietary agents for prevention and therapy of cancer
• Biochemical Pharmacology 71(2006) 1397-1421
Confusion about what to eat?
Watch 90 min video of Colin Campbell at: http://www.youtube.com/watch?v=4XzDOGZKI68&feature=related
Nutrient Density
• A nutrient density standard for vegetables and fruits: nutrients per calorie and nutrients per unit cost.
J Am Diet Assoc. 2005 Dec;105(12):1881-7.
– “Energy density and nutrient density score were negatively correlated, confirming the widely accepted notion that energy-dense foods tend to be nutrient-poor.” – “… fruits and vegetables had the highest nutrient density score because they were nutrient-rich in relation to their low energy content.”
FIGURE 4. Relation between energy density and the naturally nutrient rich score for vegetables and fruit
Drewnowski, A. Am J Clin Nutr 2005;82:721-732
Copyright ©2005 The American Society for Nutrition
Used with permission
Health Equation
• Eat foods high in nutrients and low in calories. Health = Nutrient/Calories
– – Dr. Joel Furhman Video: The Greatest Diet on Earth
• www.DrFuhrman.com
•
•
Junk Food defined
– low nutrients with high calories
Gr eens
• Greens have the highest nutrient per calorie ratio of all fruits and vegetables. – www.peertrainer.com/diet/nutrient_density.html Greens are a good source of magnesium, folic acid and vitamin K. “Vitamin K intake may be a marker of a healthy diet because it is found mainly in green, leafy vegetables”. Only 27% of people in the United States consume the average daily goal for vit K.
– Food consumption survey analysis from the Human Nutrition Research Center, part of the Agricultural Research Service. Agricultural Research/August 2007
• •
• http://www.ars.usda.gov/is/AR/archive/aug07/vitamin0807.htm
ORAC Values
• ORAC: Oxygen Radical Absorbance Capacity • Method for measuring antioxidant capacity of foods • ORAC database for 277 food items at:
– http://www.ars.usda.gov/Services/docs.htm?docid=15866
Food description
Apple juice,unsweetened without added ascorbic acid Apple, Red Delicious,raw, without skin Apple, Red Delicious,raw with skin Applesauce,unsweetened, without added ascorbic acid Artichoke (globe or French) raw Artichoke (Ocean Mist) boiled Blueberries, raw
Total-ORAC
408 2936
Calories
47 Kcal 48 Kcal
4275
52 Kcal
1965
43 Kcal
6552 9416
47 Kcal
53 Kcal
6552
57 Kcal
Data derived from: http://www.ars.usda.gov/Services/docs.htm?docid=15866
As we age, our bodies produce and accumulate a family of molecules called Advanced Glycation End-products.
Increasi ng AG Es
Gl ucos Protei n e Advanced Glycation End-product (AGE) Gl ucos e Li pi d s
Advanced Lipoxidation End-product (ALE)
Glycotoxins
Facts about AGEs
• AGEs cause proteins or lipids to be structurally altered in such a way as to make the protein or lipid dysfunctional.
– Hand analogy with cross-linking
• AGEs are proinflammatory and promote oxidative stress. • AGEs are associated with pathological conditions.
• Diabetes, renal failure, artherosclerosis, Alzheimer’s, rheumatoid arthritis, cancer,cataracts, allergic and autoimmune diseases
Food derived AGEs
• AGEs derived from food are absorbed and contribute to serum AGE levels in both animal and human studies. Ann N Y Acad Sci. 2005 Jun;1043:452-60
Increased AGEs in serum
QuickTimeª and a TIFF (Uncompressed) decompressor are needed to see this picture.
Food derived AGEs
• Once thought to only be problematic in persons with diabetes or renal failure, dietary AGEs are now being shown to cause inflammation and contribute to inflammatory and degenerative processes even in healthy individuals.
– – Diet-derived advanced glycation end products are major contributors to the body's AGE pool and induce inflammation in healthy subjects. Ann N Y Acad Sci. 2005 Jun;1043:461-6. Circulating glycotoxins and dietary advanced glycation endproducts: two links to inflammatory response, oxidative stress, and aging. J Gerontol A Biol Sci Med Sci. 2007 Apr;62(4):427-33.
Advanced glycoxidation end products in commonly consumed foods. J Am Diet Assoc. 2004
Aug;104(8):1287-91
AGE production in foods is temperature and moisture dependent.
•AGEs in a 90 gram serving of chicken breast
–9,000 kU oven frying –6,700 kU deep frying –5,250 kU broiling –4,300 kU roasting –1,000 kU boiling
Boiling, poaching, stewing vs. dry heat
So up …. .a be tter ch oic e!
Foods highest in fat contain the most AGEs, followed by meats, processed carbohydrates and then lowest in AGE content are vegetables and fruits. 1 lb = 454 grams But ter 265 kU /g Chi cken breast broi led 58 kU/ g Kel logs Rice Kri spi es 18kU /g Green beans 0. 18 kU /g Banana 0. 01 (not to scale) /g kU
J Am Diet Assoc. 2004 Aug;104(8):1287-91
Advanced glycoxidation end products in commonly consumed foods. J Am Diet Assoc. 2004
Aug;104(8):1287-91
AGEs in whole human milk 0.05 kU/ml
AGEs in Enfamil Infant Formula 4.86 kU/ml
“…since AGEs are known immune cell modulators, the introduction of infant diets, rich in AGE antigens, may account for the rise in childhood autoimmune diseases such as Type 1 Diabetes.”
Side Note
Breast-feeding reduces risk of:
Acute otitis media Non-specific gastroenteritis Lower respiratory tract infections Atopic dermatitis Asthma (young children) Obesity Type 1 and 2 diabetes Childhood leukemia Sudden infant death syndrome Necrotizing enterocolitis
Evid Rep Technol Assess . 2006 Apr;(134):1-161.
Hiding AGEs
• Unexpected places for finding food AGEs
– Soy sauce 8,700 AGE, units /15 ml
• (3 teaspoons)
– Classic Coca-Cola 8,500 AGE, units/cup – Diet Coke (soda) 9,500 AGE, units/cup
Proc Natl Acad Sci USA 1997 June 10; 94(12): 6474-6479
Regular AGE Diet
Same foods different temperatures used during preparation.
Low AGE Diet
25% decrease in GSH/GSSG
Basel ine compared to 24 months 40% increase in GSH/GSSG
45% decrease in 8-isoprostanes Increased AGER1 expression Unaltered RAGE expression Extended lifespan
(equal to mice on 40% calorie restriction)
20% increase in 8-isoprostanes Decreased AGER1 expression Increased RAGE expression Regular lifespan
Am J Pathol. 2007 Jun;170(6):1893-902. Full text available at: www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=17525257
Side note: AGEs in autism
• Proteomic studies identified a single nucleotide polymorphism in glyoxalase I as autism susceptibility factor. Am J Med Genet A. 2004 Nov
15;131(1):11-7.
– Analyses of autopsied autism brains show AGE accumulation.
• These were not brains from aged persons.
– 38% decrease in Glo1 enzyme activity in autism brains versus controls
• Results in accumulation of brain AGEs
•
Alterations of circulating endogenous secretory RAGE and S100A9 levels indicating dysfunction of the AGE-RAGE axis in autism. Neurosci
Lett. 2006 Dec 27;410(3):169-73.
– If an AGE binds to a RAGE receptor it initiates oxidative stress and inflammation. – esRAGE receptors can act as decoys to prevent AGEs from binding to the RAGE receptors. – Peripheral esRAGE levels were significantly reduced in autism.
Dietary AGEs
Self-perpetuating cycle of chronic inflammation
Adapted from a slide by Dr. Jill James (used with permission)
Heavy Metals Genetics Viral infection Inflammation GSH/GSSG
Acute Inflammation (Resolution)
PPAR-RXR dimer (downregulates transcription) TNFα IL-6 IL-1 PGJ(2) GSH/GSSG OFF-SWITCH iNOS
low
ROS
Free Radicals Transcription Cytokines factors
NFkB AP-1
NO
Nitric oxide
high
GSH/GSSG
Th1 Th2
Chronic Inflammation
Di etary AGEs Environmental insults Infection Genetic polymorphisms Dietary deficiencies
Oxi dati v e stres s
Ant ioxidan ts
Advanced glycation end products and RAGE: a common thread in aging, diabetes, neurodegeneration, and inflammation.
Glycobiology. 2005 Jul;15(7):16R-28R.
Full text available at:
http://glycob.oxfordjournals.org/cgi/content/full/15/7/16R
Break Time (5 Min)
When we return we will be discussing the “how” mechanisms of the biochemical pathways involving folate and glutathione.
The Folate Cycle
Pyrimidine synthesis Uracil Thymine
Folate sufficiency
Purine synthesis Adenine & Guanine
Folate deficiency
Methionine 10-formyl-THF 5,10-methylene THF MTHFR DHF DHFR THF B12
Methyl THF
MS
homocysteine
Double Strand Breaks
• DNA deletions • DNA translocations
The Folate Cycle
Purine synthesis Adenine & Guanine
Folate deficiency
Pyrimidine synthesis Uracil Thymine
Folate sufficiency
Methionine 10-formyl-THF DHF DHFR 5,10-methylene THF MTHFR THF B12
Methyl THF
homocysteine
MS
Methionine synthase
5,10-methylene THF MTHFR
Methyl THF
CH3THF
Methionine THF B12 MS homocysteine
Step 1:
CH3 THF
CH3 Cobalamin (B12) (Methylcobalamin)
Step 2:
CH3 Cobalamin (B12)
CH3 homocysteine (Methionine)
methionine Folate cycle SAM THF
B12
Methylation Cycle
MS
SAH homocysteine
(Methyl THF)
CH3-THF
MS : methionine synthase SAM : Sadenosylmethionine SAH : Sadenosylhomocysteine
Notes on methylation
CH3 is called a methyl group SAM is called a universal methyl donor A family of enzymes called methyl transferases serve as the working force to transfer methyl groups from SAM onto other molecules
Methyl transferase taxi analogy
Used in myelin basic protein Convert serotonin to melatonin Convert norepinephrine to epinephrine
Catecholamine
detoxification via COMT
Methylation in the liver is a detoxification pathway for many chemicals.
Gene expression
Implications for cancer
Viral silencing
Decreasing SAM/SAH ratio
Inhibits methyl transferase activity
methionine Folate cycle SAM THF
B12
Methylation Cycle
MS
SAH homocysteine
B6
CH3-THF
(Methyl THF)
cystathionine
B6
cysteine glutathione
Transsulfuration pathway
H 2O H 2O 2
Hydrogen Peroxide
Hydrogen Peroxide
+
Glutathione
Glutathione Utilization
cysteine
SOD GST
Detoxification of drugs and chemicals
H2O2
GPx
GSH
GR
H2O
GSSG
GSH: glutathione GSSG: oxidized glutathione GR: glutathione reductase GPx: glutathione peroxidase
GST: glutathione transferase SOD: superoxide dismutase H2O2: hydrogen peroxide
Notes on Glutathione
Is an antioxidant made in your body Converts hydrogen peroxide to harmless water Environmental chemicals, heavy metals, dietary deficiencies and medical drugs can deplete glutathione Glutathione has antiviral properties Lower levels found in children with autism compared to control children Protects DNA and mitochondria
What happens if hydrogen peroxide production exceeds glutathione production?
Hydrogen Peroxide glutathione Hydrogen Peroxide Hydrogen Peroxide Hydrogen Peroxide glutathione Hydrogen Peroxide
glutathione
Hydrogen Peroxide
Art by: Christy Domino
Metals Pesticides Chemicals Infections Poor diet
O2- (super oxide) OH (hydroxyl) NO (Nitric Oxide) OONO- (peroxynitrite) H2O2 (hydrogen peroxide) Damage to: DNA, proteins, lipids
oxidants
antioxidants
No oxidative stress
Free radical production in balance with antioxidants
Environmental insults Infection Genetic polymorphisms Poor diet
Oxi da tive str ess
Ant ioxidants
Environmental insults can severely tip the scale in those with increased vulnerability
• Folic acid impacts cancer risk in the following ways:
Cancer
– DNA methylation (gene expression) – DNA synthesis
• Uracil and DNA repair
– Detoxification of chemicals and control of oxidative stress
• Methylation of chemicals • Glutathione (GSH)
– GST helps inactivate chemical carcinogens into less toxic or inactive metabolites
methionine THF
B12 MS
SAM SAH H homocysteine H cystathionine cysteine L GSH
GPx GST
CH3-THF
(Methyl THF)
High and low values of ASD parents compared to controls.
SOD
H2O2
H2O
H GSSG
GR
thimerosal
Does dose make the poison
GSH
GSH
GSH
GSH
GS H
thimerosal
GS
H
GSH
GSH
Glutathione (GSH) protects against thimerosal induced apoptosis (cell death)
Genes Immun 2002 Aug;3(5):270-8
GSH
GSH
Cell Death
thimerosal
GSH
SAME DOSE of thimerosal as in previous slide!
A person with lower levels of GSH
GSH
QuickTimeª and a decompressor are needed to see this picture.
Low glutathione levels can make people more sensitive to DNA damage from a variety of mutagenic environmental exposures.
A s pecia l Th ank Yo u to Dr. J ill James Dr. P aul Cutle r My fr iend Pa ula Teri A rr anga Ray J anak And to yo u…t he pare nts !
