Introduction

At a mini-DAN! practitioner training course, one of the members of the audience asked for a "methylation for dummies" lecture. This presentation is intended to introduce the concepts of methylation and thiol biochemistry to the novice. For those more familiar with these concepts, my hope is to provide an enjoyable lecture that will be visually entertaining and integrate various pieces of the fascinating picture of autism as we currently understand it.

Basic concepts

I. Biologic systems are complex, interrelated, and synergistic. Successful treatment of children with autistic spectrum disorders requires a healthy respect for the microcosm as well as the macrocosm.

2. Autistic symptoms should not be viewed as an inevitable result of a DSM-IV diagnosis, but should generate a differential diagnosis, just as we consider potential causes of symptoms like abdominal pain.

3. Evaluation of an individual's cellular biochemistry is an emerging tool that can enable us to more effectively treat the patient and achieve better quality of life for the family.

Outline

Importance of the Gut:

Each part of the biochemistry discussed impacts the gut. The high incidence of bowel disease (both symptomatic and occult) in children with autism implies a complex interaction between cause and effect. There are both upstream and downstream effects between alterations in cellular biochemistry and gut pathology. Since the gut is full of neurotransmitters, receptors, and living organisms which generate chemicals, abnormalities in gut function affect what is designated as brain function in a less sophisticated paradigm.

Methylation:

To methylate means to add a carbon and 3 hydrogen atoms to something. This doesn't sound like such a big deal to the average person in the street, but it is actually one of the major crossroads of biochemistry and a vital process for sustaining function of many systems. The process of methylation is crucial for:

I. Making new DNA (the building blocks of life) and RNA (the blueprints and instructions for new cells)

2. Maintaining cell membranes, where signals are processed so the cell know what jobs it is supposed to do

3. Creating neurotransmitters, which carry messages to regulate emotion, attention and learning

4. Providing energy and synchrony for neuronal firing and cognitive function.

Folate Pathways:

Well functioning folate biochemistry is essential for prevention of cancers, good cardiovascular health, and normal development of the nervous system. It also plays a huge role in something charmingly called "folate dependent phospholipid methylation." In lay terms, this is a process where (more than 50 times each second!) individual cells get messages that impact the normal process of attention.

Sulfation and Metallothionein:

Sulfur biochemistry is about sticking to things we need to get rid of and escorting those things out of the body. Metallothionein likes to capture heavy metals like mercury and lead. Both are huge players in our detoxification system. This part of our biochemistry may well be "the Achille' s heel of current generations," as my friend and mentor Sid Baker likes to say. Evolutionary changes in cellular biochemistry to get rid of toxins happen at a glacial pace.  Human changes to increase our toxic burdens have happened at mach speed.

Integration of "The Biochemistry of Autism" Diagram:

The hallmark of a thoughtful approach to the child with autism is attention to the unique circumstances of the individual patient. Each part of cellular metabolism as described above has the potential to be a key to better functioning and quality of life for children with autism. To learn how requires focus, motivation, and hard work. Clearly. our patients and children are worth the effort!  Web as symbol of considering the interconnectedness of biologic systems Moving from the macrocosm to the microcosm Increasing levels of complexity .

The Chemistry of Autism Diagram

Shows the interconnectedness ofwork done by various scientists in DAN!

Lecture outline

Importance of the gut

Chandler

Genomic predisposition to inflammation

Methylation and folate pathway abnormalities leading to low glutathione

Detoxification problems

How the body makes things

In order for a substance to be converted to a product, often requires:

Feedback inhibition

.Substance -----> Product

.As product increases, it gives FEEDBACK to the substance to INHIBIT making more

Methylation Abnormalities:

Methylation is a crucial biochemical crossroads, so abnormalities here have profound consequences. Methylation provides energy and synchronicity for neuronal firing. Disorders in this pathway have profound implications for cognitive function and the ability pay attention appropriately.

Cellular consequences of reduced methylation capacity .Reduced DNA methylation

Methionine: The "queen" of essential amino acids

Methyl -CH3

Methyl groups like to be picked up: trading methyl groups is a way of changing molecular personalities

SAM

Dumps Methyl

Sam is the universal methyl donor; takes methyl transiently, then gets rid of it, forming s-adenosylhomocysteine (SAH).

Transferases: Special enzymes

COMT: Catecholamine O methyl transferase

Catecholamines are neurotransmitters which regulate "fight or flight" Activity of COMT is controlled by SAM and SAH.  Weak COMT function leads to poor methylation of catecholamines Can also affect L dopamine and steroid hormones

DPPIV = CD26 = adenosine deaminase binding protein

Adenosine deaminase

Research of Jill James PhD University of Arkansas: autistic children have significantly lower methionine, SAH, cysteine, total glutathione and "good" (reduced) glutathione; "bad" ( oxidized) glutathione is high

Methylenetetrahydrofolate reductase: MTHFR

Some common enzyme dysfunctions in autism:

Homocysteine: Earth mother who recycles and makes things

Homocysteine recycling to methionine is so important it happens 2 ways: 1) Vitamin B12 dependent methionine synthase via 5 methyl tetrahydrofolate 2) Betaine dependent homocysteine methyltransferase BHMT

Homocysteine can follow 2 paths: 1) Recycle back to methionine when all is well; 2)  Make cysteine and then glutathione under conditions of oxidant stress and high cytokines

Homocysteine is a precursor for: cysteine, taurine, and metallothionein.  Homocysteine recycling can be stalled by: High adenosine, Low P5P (also need zinc to phosphorylate B6), Poor recycling of glutathione (an area of further research)

Glutathione

Tripeptide: glutamic acid, cysteine, glycine

Reduced ( GSH) is good; oxidized ( GSSG ) is bad for you

Glutathione = A working woman who multitasks (Mnemonic: she has many vital roles but still manages to work on her gluts)

Vital roles of glutathione:

Cellular consequences of decreased glutathione: a decrease or shutdown of methionine synthase, which processes homocysteine back to methionine. Both oxidative stress and mercury shut down methionine synthase.

Systemic consequences of glutathione deficiency

OXIDATIVE STRESS WORSENS EVERYTHING

Measurable Cysteine Deficiency: common in autistic children

Oxidative Stress: potential causes includes stress, trauma, aging, infection, poor diet, pollution, toxins, drugs, radiation

Oxidative stress is analogous to RUSTING on a cellular level.  Anti-oxidants are important as scavengers of free radicals, which are loose electrons.

Methionine synthase, MB12 and homocysteine recycling are crucial for methylation and sulfation

Folate Biochemistry

Folic acid: "leaf"

Folic Acid explained in Sid Baker's book; Aka: 5-fonnyl tetrahydrofolate

Folate dependent phospholipid methylation

D4 Dopamine receptor

>50 cycles per second to focus attention

Dependent on methione synthase

Methionine synthase, AKA Mercury Sucks, because this enzyme is disabled after 60 minutes by 10 nanomolar concentrations of thimerosal

Lead at 10 micromolar concentrations reduces MS activity to 10% normal Antimony stops MS activity at nanomolar concentrations

In neurons, Methionine Synthase is activated by a signaling pathway that can be stimulated by dopamine and IGF-l. Heavy metals. especially thimerosal. inhibit this pathway and impair the activity of methionine synthase. When methionine synthase is destroyed. homocysteine increases, which increases SAH. High levels of SAH inhibit methylation, including DNA methylation which controls gene expression. Consequences are devastating.

3 Folate traps block flow of metabolic reactions and interfere with methylation cycles

Sulfation

Adding sulfate {sulfate conjugation -adding one molecule to another) is a common form of detoxification

Sulfur = "sticky;" Job is to stick to things and help you get rid of them. Sulfate can act like a switch

GAGS: glycosaminoglycans

Regulate and modulate:

Disulfide bonds maintain strength of proteins (like hair and nails) .

Cysteine-SH + HS-Cysteine <----- > Cysteine-S-S-Cysteine

Detoxification and Healing: book by Sidney M. Baker

Sulfur helps synthesize substances so they can be excreted.  Examples: sticks onto ammonia, used neurotransmitters and leftovers from our immune system.

Substances that need sulfur:

Mercury and sulfation

Interventions to support sulfation

Metallothionein: functions to grab metal ions

MetaUothionein dysfunction: consequences

Biochemical effects of Mercury Poisoning: all common in autistic children

Lab abnormalities in ASD:

Low: B6, intracellular magnesium, taurine, cysteine, Vitamin A, selenium, Intracellular folate, intracellular B12, essential fatty acids

High: opioid peptides, IgG antibodies, yeast metabolites, bacterial urinary metabolites, methylmalonic acid

Correlation of the biochemistry of autism to the gut:

Nucleotides (phosphorylated) and nucleosides: Regulate cellular perception and response. Lots of influence on the immune system, which resides largely in the gut.

G proteins: Disruption in glycoproteins leads to calcium channelopathies, abnonnalities in ion exchange, and erroneous "second messenger" information

Folate pathways: Folic acid supplementation decreases the risks of abnonnalities in epithelial tissues, like the GI tract

Methylation: Crucial for making new DNA and RNA (gut. has rapid epithelial turnover) Vital for making neurotransmitters, many of which have receptors and actions in the gut

Important for choline formation; if deficient, intestinal peristalsis is disrupted and constipation can result

Glutathione: Gut flora produces methanethiol and sulfide, which gut cells convert to thiosulfate, which ties into glutathione; Glutathione important to maintain gut epithelium; Glutathione deficiency predisposes to mercury and other toxins remaining inside cells

Metallothionein: Tries to protect intestinal brush border DPP IV from toxic elements; When metallothionein and zinc are deficient, it leads to increased copper, mercury and other heavy metals; DPP IV is disabled, which leads to: false neurotransmitters, abnormal lymphocyte activity in GI tract.  Mercury toxicity has known effects on the immune system and contributes to viral persistence.

Sulfation: Provides negative charge for function of mucin in the gut, makes a negatively charged antimicrobial barrier in the gut Activates/deactivates and protects circulating hormones and neurotransmitters: when sulfate activates CCK ( cholecystikinen) it is a signal to make oxytocin, the hormone that enables the brain to recognize social information

Breastfeeding, oxytocin, and social connectedness.  Complexities of human interactions, on a physical and emotional level Synergy of biologic systems Energy interactions.