1. Evaluation of Nutrient Levels in Children with ASD vs. Controls – Preliminary Results James B. Adams Arizona State University www.eas.asu.edu/~autism

2. Collaborators Tapan Audhya Stephen Coburn Liz Geis Julie Ingram Sanford Newmark Dena Goldberg Warren Tripp Marie Adams Funded by Autism Research Institute, Greater Phoenix Chapter of the ASA, BHARE, Arizona State University Special thanks to Vitamin Diagnostics, Doctor’s Data, and Great Plains Labs for discounted/free testing

3. Outline Vitamins and Minerals Amino Acids Essential Fatty Acids

4. Basics of Nutrition The essential major components of human nutrition are: water carbohydrates (for fuel) vitamins and minerals amino acids (from protein) essential fatty acids A deficiency of any of these results in disease, or even death in extreme cases

5. Examples of Nutritional Deficiencies Lack of vitamin C -> scurvy Lack of iron -> anemia; mental retardation in children Lack of calcium or vitamin D -> rickets Children with autism are not classically deficient, but most are low in some essential vitamins, minerals, amino acids, and fatty acids. “Children with Starving Brains” by J. McCandless

6. Why consider vitamin/mineral supplements for people with autism? 1) vitamin/mineral deficiencies common in general population 2) often restricted diets (“picky eaters”) -> limited vitamin/mineral intake 3) poor digestion (25% have chronic diarrhea, 25% chronic constipation); Endoscopies by Wakefield, Buie, Krigsman show damage to gut -> limited absorption of nutrients? 4) Rosseneu, Shaw research: overgrowth of harmful bacteria in GI tract: fewer “good” bacteria which actually produce several vitamins 5) 18 studies demonstrated benefits of vitamin B6/Mg 6) 1 study showed vitamin C was beneficial in autism 7) Prof. Megson: cod liver oil (with high levels of vitamin A, D, and essential fatty acids) helped with gaze aversion, behavior 8) Dr. Rimland’s set of parent-reported case studies on benefits of vitamins/minerals

7. Goals: 1) Evaluate nutritional status of unsupplemented children with ASD vs. unsupplemented typical children, age and gender matched. 2) Check for correlations of symptoms of autism with nutritional levels Participants: Enrolled: 48 ASD, 35 controls Age: 3-9 years All children from Arizona. No use of vitamin/mineral supplements in 2 months prior to sample collection.

8. Tests Vitamins, minerals, essential fatty acids, amino acids (blood, urine) – Vitamin Diagnostics Vitamin B6 variants and enzymes – Stephen Coburn (Purdue Un.) Toxic elements and essential minerals in hair – Doctor’s Data Urinary Organic Acid Testing – Great Plains Lab Dietary Assessment of Nutrient Intake (1 week food diary) All samples sent blinded to labs.

9. Vitamins Ref Range (RR) defined as between bottom 15% and top 15% Only small differences in vitamin levels between autism and controls (not statistically significant)

10. ARI Survey of Parent Ratings of Treatment Efficacy % Worse % No Change % Better Number of Reports Vitamin A2%58%41%618 Folic Acid3%54%42%1437 Vitamin B34%55%41%659 Vitamin B124%33%63%192 Vitamin C2%57%41%1706

11. Why do children with autism benefit from vitamins if levels not very different from typical children? 1)Typical children do not eat 5 servings of fresh fruit and vegetables each day, so the level of vitamins in typical children is not optimal. 2)Several studies show children with autism are under increased oxidative stress, suggesting a need for increased levels of antioxidants.

12. Vitamin B6 and Magnesium Over 20 studies, including 11 double-blind, placebo-controlled studies, found that high dose vitamin B6 (8 mg/pound bodyweight) with Magnesium (3-4 mg/pound bodyweight) resulted in wide range of behavioral improvements Only 2 negative studies: one with very few subjects, and one with half dose Overall, very safe, and helps 50% of children and adults.

13. Vit B6 – Coburn data Urinary excretion of pyridoxic acid is low in autism Oxidase activity low but not significant

14. Vitamin B6 levels First Study Autism (n=35)56 +/- 13 ng/ml Controls (n=11)36 +/- 9 ng/ml ttest = 0.00002 Second Study Autism (n=47)62 +/- 41 Controls (n=33)53 +/- 27 not significant Combined First and Second Study Autism (n=82)72 +/- 32 Controls (n=44)52 +/- 25 p= 0.05 Conclusion: Some children with autism have unusually high levels of B6, and a few have unusually low levels of B6 - Explanation: study by Dr. Tapan Audhya found that 4 enzymes for conversion of B6 are defective in autism, such that conversion rate is much lower than normal

15. Substrate Requirement for Maximal Activity of P5P-Dependent Enzymes Autistics often have weak B6-dependent enzymes, so may need very high B6

16. Substrate Requirement for Maximal Activity of P5P-Dependent Enzymes Some autistics need high-dose B6 to make important neurotransmitters

17. Autism Treatment Study: Effect of B6 (10mg/kg/day) + Zn (25mg) + Mg (400mg) on Kryptopyrrole Levels Conclusion: High-dose B6 very helpful for treating pyroluria

18. High-dose B6 + multivitamin treatment study (Audhya) 184 Volunteers 1. Unsuccessful with Diet Intervention 2.Given Daily one centrum advanced formula Multivitamin Tablet with food, 25 mg Zn ++ (as Gluconate and Citrate) and 400 mg Mg ++ (as Gluconate & Oxide) 128 (A) (95:33) Pyrindoxine - HCℓ 5 mg/kg/day 56 (B) (41:15) Pyridoxine – HCℓ & Pyridoxal – PO 4 1 mg +4.7 mg/kg/day

19. Outcome of the study 89 Volunteers Substantial Improvement; Almost normal 86 VolunteersNo perceptible change; Behaviorally or physically 9 Volunteers No improvement; and additional adverse reaction

20. ARI Survey of Parent Ratings of Treatment Efficacy % Worse % No Change % Better Number of Reports Vitamin B6 alone 8%63%30%620 Magnesium6%65%29%301 Vitamin B6 with Magnesium 4%49%47%5780 P5P (Vit. B6)13%37%51%213

21. Summary re. Vitamin B6 Some children and adults with autism benefit from high- dose B6, to make neurotransmitters, glutathione, and many other important substances Simple measurement of kryptopyroles in urine determines if high-dose vitamin B6 (or zinc) is needed (caution – destroyed by light, so collect in dark). Doses up to 10-15 mg/kg may be needed (1000 mg max); takes about 6 months to lower kryptopyroles in urine Always give at least half as much magnesium (400 mg max).

22. Vitamin B12 Methyl cobalamin (methyl form of vitamin B12) is most active form of vitamin B12 Vitamin B12 is very hard for body to absorb orally Injectable form of methyl-B12 may help up to 70% of autistics per informal reports by Dr. Jim Neubrander, at doses of 75 mcg/kg bodyweight, 2x/week, for 6+ weeks Safe, but needs research to verify effectiveness Research by Prof. Richard Deth shows it is needed to produce glutathione, and to help Dopamine 4 receptor function (important for attention)

23. Low Cysteine and Glutathione Studies by Jill James and Tapan Audhya find low cysteine and extremely low glutathione in children with autism; glutathione is important antioxidant and for detoxification Treatment study by Jill James finds that 800 mcg of folinic acid and 1000 mcg of TMG over 8 weeks improve cysteine and glutathione levels somewhat Addition of 75 mcg/kg of methyl B12 injections helps more Informal reports of improvement in behavior

24. Minerals Zinc slightly high in whole blood, but average in RBC Copper results mixed – slightly high in RBC, slightly low in serum, not statistically significant Disagrees with results by Walsh (possibly due to LabCorp Ref Range and age of controls?)

25. Minerals (cont.) Iron: slightly slow in serum, but serum ferritine is more relevant and is “normal” However, many children with exceptionally low serum ferritine (10 with < 10 mcg/l) Calcium: low in urine, possibly due to low intake (GFCF diet)

26. Toxic Elements in Hair Most toxic elements are lower in autism, suggesting an overall problem with excretion; aluminum, tin, and possibly nickel are statistically significant

27. Essential Minerals in Hair Manganese, molybdenum, and lithium are low in hair. Manganese slightly low in blood, partially consistent with hair. Low lithium is consistent with previous study.

28. Essential Minerals in Hair (cont.) Low iron and possibly low cobalt. (iron not reliable in hair) Slightly low selenium. Possibly slightly high sulfur.

29. Lithium – previous ASU study The only abnormality in mothers of children with ASD was low levels of lithium: all ages: -40%, p=0.05 mothers of children ages 3-8: -56%, p=0.005 (highly significant!) Similarly, children with ASD had lower levels of lithium all ages: -15%, not significant ages 3-6 yr: -30%, p=0.04

30. Importance of Lithium Hair is a reliable measure of lithium Lithium is probably an essential mineral (not well studied) Study of goats on lithium-deficient diet found: –decreased activity of monoamino oxidase, which is of particular importance to manic-depression, chronic schizophrenia, and unipolar depression. –lowered immunological status, and suffered from more chronic infections (may explain why children with autism had more ear infections) Lithium concentrations highest during first trimester, and highest in the brain, so a deficiency of it could affect early fetal development, including early brain development

31. Lithium - continued Several studies have show low lithium correlates with: schizophrenia neurosis suicide behavior problems crime (homicide, rape, burglary, theft, drug use, juvenile runaways) Finally, a four-week placebo-controlled study of 24 former drug users found that 400 mcg/day of lithium resulted in steady increases in mood scores, especially in subcategories reflecting happiness, friendliness, and energy.

32. Lithium Not included in most nutritional supplements, or in prenatal supplements An estimated RDA is 1000 mcg/day, and people in the US consume only about 500 mcg/day Extremely high doses of lithium (1,000,000 mcg/day) are used as a psychiatric medication, primarily for “calming/mood stabilization”, especially for bipolar disorder; nearly toxic at that dose RECOMMENDATION: a dosage of 200-1000 mcg/day should be safe, and may be beneficial to younger children with autism and their mothers More research needed

33. Essential Minerals - Iodine Iodine: 45% lower in ASD than controls, p=0.005 (highly significant!) in 3-6 yr old group, similar value (-47%) Caution: no data showing that iodine in hair correlates with level in body (blood is standard measurement) iodine is an essential mineral major role of iodine in body is in thyroid function a deficiency of iodine causes goiter (enlarged thyroid) and mental retardation (Cretinism) worldwide, the leading cause of mental retardation is iodine deficiency, affecting roughly 20 million children

34. Iodine - continued In early 1900’s, iodine deficiency was up to 30% in some parts of the US iodine in salt is believed to be sufficient to make iodine deficiency very rare in the US/western world however, iodine levels in blood have declined 50% from 1970’s to 1990’s per NHANES I and III, possibly due to decreased salt intake many fast foods (fries, chips) use non-iodinized salt CAUTION: too much iodine can also decrease thyroid function RECOMMENDATION: measure iodine in blood, supplement at modest level if low

35. ARI Survey of Parent Ratings of Treatment Efficacy % Worse % No Change % Better Number of Reports Calcium E :2%62%36%1378 Magnesium6%65%29%301 Zinc2%51%47%1244

36. Summary of Minerals Low iron in some children – supplement only if low. Low calcium in some children (esp. if dairy-free) Low lithium in children and mothers, which can affect behavior Low iodine is a leading cause of mental retardation, and should be supplemented Low toxic metals in hair suggests problem with excretion of toxic metals.

37. Research Study of Multivitamin/mineral supplement 3 month study of Spectrum Support (by Brainchild Nutritionals) Double-blind, placebo-controlled Dosage is slowly increased to maximum over first 2 months, then held constant parent ratings of changes small study - 20 children only

38. Vitamin C results (at end of study) rangeaverage Placebo: 0.9-1.4 1.03 Suppl.1.0-2.01.33 typical child (age 3-8)1.45 Placebo children are 2 standard deviations below average value 500 mg raised vitamin C to near-normal levels, but more may be better (1000 mg)

39. Overall Results Based on parent evaluations on final day of study 7 point scale 1=much worse 2=worse 3=slightly better 4=same 5=slightly better 6=better 7=much better

40. Overall Results CategoryPlaceboSupplementDifference Sociability5.15.3+0.1 Expr. Language5.65.9+0.3 Rec. Language4.95.8+0.9 Eye Contact4.95.5+0.6 General Behavior4.35.1+0.8 Sleep3.95.4+1.5 Gastrointest.3.95.4+1.5 Overall5.15.5+0.4 Sleep and GI results are statistically significant (p<0.005); other results positive, and worth further investigation

41. My recommended daily dose per 20 pounds of bodyweight, up to 100 pounds (i.e., for a 60 pound child, multiply by 3) Take with food, split into 3 doses Vitamin A1000 IU Vitamin B17.5 mg Vitamin B27.5 mg Vitamin B325 mg (10 mg niacin, 15 mg niacinamide) Vitamin B520 mg Vitamin B680 mg Vitamin B12200 mcg Folic Acid50 mcg Folinic Acid100 mcg Biotin100 mcg Choline70 mg Inositol30 mg PABA5 mg Vitamin C300 mg Vitamin E60 mg Mixed Carotenes3000 IU Chromium20 mcg Copper 0 mg (most autistics don’t need) Iodine30 mg Lithium0.2 mg Magnesium120 mg Manganese1 mg Molybdenum40 mcg Selenium25 mcg Vanadium13 mcg Zinc5-15 mg Calcium: Dosing independent of bodyweight: 400 mg for ages 2-5, 500-600 mg for older children Potassium from fruits and vegetables (esp. potatoes, avocados), esp. for low muscle tone

42. Recommended Vitamin/Mineral Supplements Kirkman’s Super Nu Thera – the original high-B6/Mg supplement for autism; contains many vitamins/minerals, but not a complete formulation; www.kirkmanlabs.com Kirkman’s Spectrum Complete: broad-spectrum formulation, with moderate B6 Brainchild’s Spectrum Support: broad-spectrum liquid vitamin/mineral supplement with moderate B6; www.brainchildnutrionals.com www.brainchildnutrionals.com D-Plex: broad-spectrum vitamin/mineral supplement with some amino acids; www.danplex.comwww.danplex.com Awaken Nutrition: broad-spectrum liquid multivitamin/mineral with high B6; www.awakennutrition.com www.awakennutrition.com Need extra Calcium with most of above products.

43. Amino Acids Protein is composed of long strands of many amino acids The body needs to digest protein to small peptides or individual amino acids Amino acids can be measured in plasma (fasting) or urine (24-hr best) Caution – high level in urine sometimes indicates wasting, resulting in low levels in body

44. Essential Amino Acids in Urine Lysine high and possibly significant; Methionine slightly low but not significant; Histidine slightly high but not significant Note: high level in urine may mean low level in body

45. Conditionally Essential Amino Acids in Urine High glycine and high cystine in urine suggests low level in body; Worrisome that cystine excretion is high

46. Metabolic Amino Acids in Urine Taurine and beta aminoisobutyric high but not statistically significant; high taurine probably suggests wasting, and hence a need for taurine

47. Summary of Amino Acids Some children with autism have low levels of amino acids in body Recommendation: measure levels, and supplement if low Also, consider measuring neurotransmitter levels in platelets, and then supplement with precursors if low (neurotransmitters made from amino acids) – more research needed

48. Essential Fatty Acids Essential Fatty Acids are termed “essential” because they are necessary for human life. The major types of essential fatty acids are Omega-3 and Omega-6. Ancient human diets contained a roughly 1:1 ratio of Omega-3 to Omega-6 fatty acids. However, since Omega-3 fatty acids spoil much faster than Omega-6, commercial food processors usually remove them or “hydrogenate” them to increase shelf life. Today, American diets contain a roughly 1:15 ratio of Omega 3 to Omega-6; in other words, most people in the US are very low in Omega 3 fatty acids.

49. EFA’s in Autism A recent study in France found that children with autism had normal levels of Omega-6, but their levels of Omega-3 were 25% below “normal”, and even “normal” is probably far from “optimal”. A small preliminary study by Gordon Bell (Un. Stirling) of 7 children with ASD also found less EPA and DHA, and sometimes more arachidonic acid, than control subjects.

50. Sources of Omega-3 Fatty Acids In human diets, the major source of omega-3 fatty acids is fish. Fish contain two important types of Omega-3 fatty acids, EPA and DHA. Some fish contain much more omega-3’s than others.

51. Importance of EPA and DHA EPA is important in reducing inflammation (such as in the GI tracts of children with autism). DHA is critical to brain development, and 20% of an infant’s brain is made of DHA. Both are critical to all cell membranes in the body; they regulate nutrients going into the cell, and waste leaving the cell. They also affect the release and reuptake of neurotransmitters.

52. ω 3 ω 6 Dry Skin Excessive Thirst & Sweating Impairment of Vision Frequent Urination Sticky Platelet Dandruff Tingling in Arm & LegKidney Degeneration Learning Disability Drying up Glands Motor In-coordination Dry, Dull and Loss of Hair High Blood Pressure Rough & Dry Skin Mental Deterioration Susceptibility to Infection Immune DysfunctionMale Sterility/Miscarriage Heart/Circulatory Problem Growth Retardation Poor Wound Healing Behavioral Disturbance Common Symptoms of Essential Fatty Acid Deficiency

54. Treatment with EFA’s Supplements of fish oil have recently been shown to be effective in treating a range of psychiatric illnesses, including: schizophrenia depression bipolar ADD/ADHD Several physicians are using them to treat autism, with some good anecdotal results, but there has not yet been a formal study of EFA supplements in people with autism.

55. Essential Fatty Acid Study – 2 month Double-Blind, Placebo Controlled Phased in gradually over 2 weeks, continued for 6 weeks Ages 3-6: 2 gel caps per day Ages 7-12: 3 gel caps per day Ages 13 and over: 4 gel caps/day ProOmega: 175 mg EPA, 125 mg DHA, 50 mg other omega 3 per capsule ProDHA: 250 mg DHA, 100 mg EPA, 50 mg other omega 3 per capsule

56. Results - Total ATEC Score StartEndDifference ProDHA66.758.1-8.6 ProOmega55.952.8-3.2 Placebo64.857.7-7.1 ProDHA group improved slightly more than Placebo, primarily due to slightly improved Sociability ProOmega was slightly worse than placebo

57. Who improved? Changes of more than 10 points occurred in: ProDHA: 8 of 24 improved, 0 worse ProOmega: 3 of 31 improved, 4 worse Placebo: 3 of 26 improved, 0 worse In the ProDHA group, the responders consumed only 1.5 servings of seafood/month, vs. 5 servings/month in non-responders. So, people with low seafood consumption are more likely to be low in Omega 3’s, and hence more likely to benefit from supplement

58. Sociability Subscale of ATEC

59. EFA Study – 9 months Pre/post measurements of EFA levels in children with autism vs. typical children Open design (no placebo group) Only treating those children with low EFA levels

60. Control (n=42) Autistic (n = 67) ά-Linolenic (18:3n3)1.4 - 4.1 1.0 – 5.2 Eicosapentaenoic (20:5n3)1.8 - 20.2 1.3 - 15.9 Docosapentaenoic (22:5n3)26 - 69 21.5 - 47.6 Docosahexaenoic (22:6n3)34 - 106 25.2 - 78.7 Linoleic (18:2n6)70 – 150 93 - 220 Gamma Linolenic (18:3n6)0.9 – 2.8 2.3 – 5.6 Dihomo Gamma Linolenic (20:3n6)20 – 39 31 - 67 Arachidonic (20:4n6)120 - 235 167 - 260 Docosadienoic (22:2n6)0.5 – 2.0 0.9 - 3.8 Docosatetraenoic (22:4n6)8.5 – 106 7.2 – 95.4 Erucic (22:1 n12)2.2 – 9.6 1.7 – 11.9 Nervonic (24:1 n15)32 - 68 27.6 – 75.7 Arachidic (20:0)1.4 – 10.8 1.0 – 8.3 Behenic (22:0)5.5 – 32.0 4.3 – 40.6 Hexacosanoic (26:0)1.0 - 8.9 3.9 – 21.3 Fatty Acid Level in Autistic Populations (nmole/ml) 7 – 17 years; mean 14 years

61. Biochemical Effect of Fatty Acid in Autistic Children N= 67 (47 boys, 20 girls)7 – 17 years age Dose Given 2g ω 3 (EPA:DHA= 1:1.63) + 0.5 g ω 6 + 400 mg Ca ++ with Vit- D Polyunsaturated/Saturated fat Ratio = 4.8 Duration of study 9 – 11 months. Blood Pressure 4.8/2.2 – 6.1/3.0 mm Hg Heart Rate 6 – 14 Beats/Min Norepinephrine 18 – 26% Prostacyclin-I-(PGI 2 ) ( Vasodilation) 14 – 21% Thromboxane A 2 (Pro-Coaglulation) 8 – 15% Leukotrienes (Pro-Inflammatory) 19 – 30% HDL 11 – 20% LDLNo Change

62. Control Autistic Autistic + EFA ( n=42) (n=67) (n =26) Eicosapentaenoic (20:5n3)1.8 - 20.2 1.3 - 15.9 2.2 – 28.7 Docosapentaenoic (22:5n3)26 - 69 21.5 - 47.628.9 – 86.7 Docosahexaenoic (22:6n3)34 - 106 25.2 - 78.749 - 132 Dihomo Gamma Linolenic20 – 39 31 - 67 26 – 49 (20:3n6) Arachidonic (20:4n6)120 - 235 167 – 260132 - 206 Fatty Acid Level in Autistic Populations (nmole/ml) 7 – 17 years; mean 14 years

63. Parental Assessment after Treatment with Fatty Acids Symptoms Improvement Abnormal Bowel movement 80% Improvement (63/63) Cognitive and Motor Skill25% Improvement (20/63) Hyper Irritability 35% Improvement (22/63) Limited Eye Contact40% Improvement (22/63) Social Withdrawal 33% Improvement (30/63) Short Attention Span 20% Improvement (10/63) Repetitive Movements, such as, Rocking 35% Improvement (63/63) Speech RegressionSignificant improvement (48/63) Sleep Patterns50% Improvement (32/63)

64. Biol Psychiatry. 2006 Aug 22; [Epub ahead of print] Omega-3 Fatty Acids Supplementation in Children with Autism: A Double-blind Randomized, Placebo-controlled Pilot Study. Amminger G. Berger GE, Schafer MR, Klier C, Friedrich MH, Feucht M. Department of Child and Adolescent Neuropsychiatry (GPA, MRS, CK, MHF, MF), Medical University of Vienna; Vienna, Austria; and ORYGEN Research Centre (GPA, GEB), University of Melbourne, Melbourne, Australia. BACKGROUND: There is increasing evidence that fatty acid deficiencies or imbalances may contribute to childhood neurodevelopmental disorders. METHODS: We conducted a randomized, double-blind, placebo-controlled 6- week pilot trial investigating the effects of 1.5 g/d of omega-3 fatty acids (.84 g/d eicosapentaenoic acid,.7 g/d docosahexaenoic acid) supplementation in 13 children (aged 5 to 17 years) with autistic disorders accompanied by severe tantrums, aggression, or self-injurious behavior. The outcome measure was the Aberrant Behavior Checklist (ABC) at 6 weeks. RESULTS: We observed an advantage of omega-3 fatty acids compared with placebo for hyperactivity and stereotypy, each with a large effect size. Repeated-measures ANOVA indicated a trend toward superiority of omega-3 fatty acids over placebo for hyperactivity. No clinically relevant adverse effects were elicited in either group. CONCLUSIONS: The results of this study provide preliminary evidence that omega-3 fatty acids may be an effective treatment for children with autism.

65. Conclusion Most autistic children have low level of ω 3 fatty acids. Some autistic children have high level of ω 6 fatty acid. The ratio of ω 3 /ω 6 can be altered by administration of ω 3 & ω 6 fatty acid (4:1 ratio) for 9 -10 months. Younger children need a ratio higher in DHA (for growth of neurons), and older children/adults need more EPA

66. Recommendation Measure EFA levels in RBC membranes, especially if GI or sleep problems Supplement with 1-2 g/day of omega’s from fish oil and 250-500 mg/day of omega 6’s in borage oil (younger children at lower dose, teens/adults at higher dose) Also give calcium to improve absorption (500 mg/day)

67. ARI Survey of Parent Ratings of Treatment Efficacy % Worse % No Change % Better Number of Reports Fatty Acids2%42%55% 626

68. Summary Vitamins – little difference (but probably needed) Vit B6 – many studies show that high-dose B6 helps some children and adults Minerals – many children need calcium supplement esp. if GFCF –iron levels similar to “typicals”, but many need iron –Lithium supplement may help many children Amino Acids – measure levels of amino acids and possibly neurotransmitters, and give customized supplement if needed Hair: low toxics in hair, suggesting poor detoxification Fatty Acids – most children with autism need fish oil (omega 3) with some omega 6 (evening primrose oil or borage oil)

69. Correlations with ATEC - total Correlations are marginally significant at.25 or greater (p=0.1) Carotenes-0.52 Calcium-0.26 GABA (urine)-0.24 Phosphoserine (urine)0.30 Arsenic (hair)0.29 Manganese(hair)0.46 Caution: Correlation is not necessarily causation!

70. Correlations with ATEC – Speech subscale Carotenes -0.40 Serum Ferritine-0.25 Hematocrit-0.28 Amino Acids (urine) Valine0.28 Leucine0.28 Phenylalanine0.30 Glutamine0.28 Alanine0.28 Phosphoserine0.28 Manganese (hair)0.25

71. Correlations with ATEC – Sociability Subscale Carotenes-0.43 Niacin-0.28 Hematocrit-0.26 Serum Iron-0.32 GABA-0.26 Phosphoserine0.25 Manganese (hair) 0.39

72. Correlations with ATEC – Sensory/Cognition Subscale Carotenes -0.47 Niacin-0.25 Serum Ferritin-0.31 Hematocrit-0.33 GABA-0.26 RBC Copper0.32 Phosphoserine0.25 Arsenic (hair)0.29

73. Correlations with ATEC Health Subscale Carotenes-0.33 Serum Molybdenum-0.26 Sulfur (hair)-0.30 Manganese (hair)0.44

74. Conclusion of Statistical Analysis Many measurements correlate with severity of autism, and with subscales of severity –Caution: need to replicate with larger study

75. Conclusion Children with autism are “Children with Starving Brains and Starving Bodies” Many need essential vitamins and minerals Some need some amino acids Many need essential fatty acids Balanced diets rich in vegetables, fruits, and protein are needed Supplements also needed in most cases, and have proven benefit