This is an exploration post on three autism microbiome samples (all were processed on Thryve, two had their FASTQ files processed on BiomeSight.com for a second interpretation of the bacteria present). I will follow the analysis pattern that I used for myself in this post, Microbiome Outliers.

The three samples are:

• A – Autism
• B – Autism
• C – Diagnosed as ASD, main characteristic is speech delay (been there, done that personally)
• D – Diagnosed as ASD, then PANS/PANDA and possible Mast Cell issues. Unfortunately – this was done by BiomeSight and not Thryve, so not exactly comparable (See The taxonomy nightmare before Christmas…)

Remember this is experimental, possibly 10 years ahead of conventional medicine – and will likely be subject to evolution. It is the best that I am aware of (and gladly be informed of better!). It will give three ‘samples’ to compare other autism microbiome against.

Core Supplements Suggested

There appears to be no shared pattern here. I usually look at items below 5% or above 95% (Outliers). What is an outlier for one child is not an outlier for the other child.

For D, the low DAO production could present appearing to be a Mast Cell issue (That is not an actual issue with Mast Cells, instead low DAO)

End Product Outliers

Again, we find no shared patterns.

We see 3 children share low Indole levels. Note:  The amino acid tryptophan is an indole derivative and the precursor of the neurotransmitter serotonin.^[2]

KEGG Module Outliers

To put this simply, Modules are a complex of processes.

Kegg Enzyme Outliers

This looks at enzymes produce by the bacteria. There are many, many enzymes – far more than is commonly known.

Here we have a reversal between A and B. Above, B shows few abnormalities and A shows many abnormalities. With the enzymes, B jumps ahead of A by a long distance! B has 453 items, A just 51.

There were no items that both had.

It is interesting to note that both children with ASD have very short lists here.

Bottom Line

Until I hit the Enzymes, I was coming to the impression that A has a jacked microbiome and B did not. That was suddenly changed when I looked at the Enzymes, B had a massive number of different shifts.

C and, who has an ASD diagnosis with speech issues being the main issue (not behavior issues etc). We see just 2,5 (versus 53 versus 451) enzyme outliers using Thryve data. This hints at enzyme production dysfunction by the microbiome as an area of interest for more severe autism.

Note: Getting suggestions from enzyme outliers or KEGG Module are features on my backlog. It will be complex to implement well.

If you look at my own latest results, Microbiome Outliers, the only thing that I had was “Rare” which could be ascribed to noise (incompleteness) of the lab results.

I did look at their results with BiomeSight processing too. They are roughly similar (we have a lot less samples – less than 10% of samples, so those results are more volatile/unreliable to compare against our sample population – Thryve is the better choice of lab solely because we have more samples uploaded. Please process their FASTQ files thru biomesight.com to build our sample population ).

Post coming soon a three way family comparison – Parent, ASD child, non ASD child. Drilling down across all data. It will be a long post.

This is a continuing series of blogs looking at OATS results and autism. The goal is to filter out items that are relevant to autism. The series is not intended to do a full explanation of the OATS test or a single person’s result. The OATS test is not autism specific but general health (often IBS/FM/CFS focuses).

Remember that our knowledge is constantly changing (unfortunately most MDs knowledge of the literature is stale).

Panel A

• 3-Hydroxybutyric
  • “Furthermore, levels of 3-hydroxybutyric acid and melatonin were significantly lower [in ASD] ” [2020] [2018] In this case not.
• Acetoacetic – nothing explicit [2018][2019]
• 4-Hydroxybutyric (gamma-hydroxybutyric acid) – high levels may be associated to specific DNA. [2019] [2016] [2009]
• Ethylmalonic
  • ” ethylmalonic acid (P = 0.043) were significantly elevated in individuals with ASD.” [2020] – in this case not
• Methylsuccinic – nothing found
• Adipic – again, this child is normal range
  • ” the increase in adipic acid concentration was significantly and indirectly correlated with the severity of the deficit in socialization and communication skills in children with an ASD” [2016]
  • ” Overall, no increase in the concentration of adipic acid in children with ASDs compared to TD children, however when considering vitamin B supplementation in ASD there were significantly increased level of urinary adipic acid in children with an ASD not taking vitamin B supplementation compared to supplemented children or to TD children. ” [2016]
  • B vitamin supplementation reduces excretion of urinary dicarboxylic acids in autistic children [2011] B vitamins lowers
  • Significant differences were found between the autistic children and the control group in organic acids: 2-oxoglutaric, isocitric, citric, 4-hydroxybenzoic, 4-hydroxyphenylacetic, hippuric, adipic, suberic (all with p<0.05). [2011]
• Suberic
  • “No significant difference were observed in suberic acid. ” [2016]
• Sebacic – nothing found

Panel B

• Vitamin B12
  • “For example, levels of vitamins B₁, B₆, B₁₂, A and D are often reported to be low in ASD children. ” [2020]
• Vitamin B6
  • “For example, levels of vitamins B₁, B₆, B₁₂, A and D are often reported to be low in ASD children. ” [2020]
• Vitamin B5
  • “Based on semi-blinded assessment, the treatment group, compared to the non-treatment group, had significantly greater improvement in autism symptoms and developmental age. The treatment group had significantly greater increases in EPA, DHA, carnitine, and vitamins A, B2, B5, B6, B12, folic acid, and Coenzyme Q10. ” [2018]
• Vitamin B2
  • “Based on semi-blinded assessment, the treatment group, compared to the non-treatment group, had significantly greater improvement in autism symptoms and developmental age. The treatment group had significantly greater increases in EPA, DHA, carnitine, and vitamins A, B2, B5, B6, B12, folic acid, and Coenzyme Q10. ” [2018]
• Vitamin C
  • The risk for scurvy in children with neurodevelopmental disorders [2020] – “Most affected children had a previous diagnosis of a medical or developmental condition (especially autism spectrum disorder)”
  • Scurvy as a Sequela of Avoidant-Restrictive Food Intake Disorder in Autism: A Systematic Review [2020]
  • Associations Between Broader Autism Phenotype and Dietary Intake: A Cross-Sectional Study (Japan Environment & Children’s Study) [2020] “consumed less vegetables, fruits, and fish and shellfish, and they consumed lower folate, vitamin C, vitamin D, and n-3 PUFA than their counterparts.”
• CoQ 10
  • “Based on semi-blinded assessment, the treatment group, compared to the non-treatment group, had significantly greater improvement in autism symptoms and developmental age. The treatment group had significantly greater increases in EPA, DHA, carnitine, and vitamins A, B2, B5, B6, B12, folic acid, and Coenzyme Q10. ” [2018]
• NAC
  • “We concluded that N-acetylcysteine is safe and tolerable, reduces hyperactivity and irritability and enhances social awareness in children with autism spectrum disorder. ” [2020]
• Biotin
  • “Low levels of biotin, plasma glutathione, RBC SAM, plasma uridine, plasma ATP, RBC NADH, RBC NADPH, plasma sulfate (free and total), and plasma tryptophan; also high levels of oxidative stress markers and plasma glutamate.” [2011]

This is a continuing series of blogs looking at OATS results and autism. The goal is to filter out items that are relevant to autism. The series is not intended to do a full explanation of the OATS test. The OATS test is not autism specific but general health (often IBS/FM/CFS focuses).

Remember that our knowledge is constantly changing (unfortunately most MDs knowledge of the literature is stale). In going thru the literature, remember what is reported is the result of a group and may not apply to a specific child with autism. Each child has difference symptoms and thus different microbiome dysfunction.

The purpose of the literature review is to identify items that are recognized as autism associated and to know what to follow up on with your medical professional.

This article, A Metabolomics Approach to Screening for Autism Risk in the Children’s Autism Metabolome Project [2020] is worth a read.

Panel A

Glyceric

Nothing found

Glycolic

A low value is seen here and in disagreement with the literature

•  Other metabolites increased are 3,4-dihydroxybutyric acid, glycolic acid and glycine, cis-aconitic acid; phenylalanine, tyrosine, p-hydroxyphenylacetic acid, and homovanillic acid [2014]

Oxalate

A high value is seen here and in agreement with the literature

• Children with ASD demonstrated 3-fold greater plasma oxalate levels compared with reference and 2.5-fold greater urinary oxalate concentrations [2012]
• Low oxalate diets have been tried with no published results [2020]

Lactic

Appears to be connected to anxiety. High levels are associated with brain fog and loss of executive function with other conditions. Child is high.

• Brain lactate as a potential biomarker for comorbid anxiety disorder in autism spectrum disorder-reply [2015]
• The relationship between lactate level and the repetitive behavior domain of the Autism Diagnostic Interview-Revised was statistically significant. [2020] See this post for possible ways to reduce..

Pyruvic

Child is upward trending. The literature refers to the ratio being high, and in this case, the ratio appears to high. An appropriate goal would be to reduce lactic acid.

• Higher Lactate Level and Lactate-to-Pyruvate Ratio in Autism Spectrum Disorder [2020]
• A significant elevation was observed in the levels of NO, MDA, protein carbonyl, and lactate to pyruvate ratio in the plasma of Omani autistic children as compared to their age-matched controls. [2012]

Panel B

Succinic

We see low values, the literature reveal that this may be connected to some SNPs in some.

• Bi-allelic Mutations in ALDH5A1 is associated with succinic semialdehyde dehydrogenase deficiency and severe intellectual disability [2020] [2020]
• Succinic Semialdehyde Dehydrogenase Deficiency Presenting as Autism Spectrum Disorder [2016]
• Safety and efficacy of oral DMSA ( dimercapto succinic acid ) therapy for children with autism spectrum disorders: part B – behavioral results [2009] “DMSA had significant improvements on all the assessment measures.”
• “DMSA greatly increased the excretion of lead, substantially increased excretion of tin and bismuth, and somewhat increased the excretion of thallium, mercury, antimony, and tungsten. ” [2009]

Fumaric

Skipping – not abnormal and nothing clear in the literature.

Malic

This person is high. Nothing found in the literature.

2-Oxoglutaric

This person is slightly low.

This lead to a single study from [2011]: “Significant differences were found between the autistic children and the control group in organic acids: 2-oxoglutaric, isocitric, citric, 4-hydroxybenzoic, 4-hydroxyphenylacetic, hippuric, adipic, suberic (all with p<0.05).”

There was also “The ASD group had higher levels of phenylactic acid but decreased amounts of aconitic acid, phosphoric acid, 3-oxoglutaric acid, and carboxycitric acid compared to TD children. ” [2019]

Aconitic

This person is low.

This was also found low in the last study “decreased amounts of aconitic acid” [2019] and contrary results in ” Interactions among diet, intestinal flora and genes may explain such findings… Other metabolites increased are 3,4-dihydroxybutyric acid, glycolic acid and glycine, cis-aconitic acid; phenylalanine, tyrosine, p-hydroxyphenylacetic acid, and homovanillic acid are all involved in the tyrosine pathway leading to neurotransmitter cathecolamine.” [2014]

Citric

This person is slightly low.

“Three of 7 patients demonstrated abnormalities in citric acid metabolites, bacterial metabolism, and fatty acid oxidation markers. ” [2015] This implies abnormalities may only occur in a subset.

3-Methylglutaric

This person is slightly elevated.

” Metabolomic analysis of urinary organic acids revealed that three metabolites, 3-hydroxy-3-methylglutaric acid (P = 0.008), 3-methyglutaconic acid (P = 0.018), and ethylmalonic acid (P = 0.043) were significantly elevated in individuals with ASD.” [2020]

3-Hydroxyglutaric

Nothing found in the literature. Person is in normal range.

3-Methylglutaconic

Nothing found in the literature. Person is in normal range.

Bottom Line

We have done another section of the OATS report. Correcting by diet or drugs some of these shifts is another level of analysis; one that too-often there is little information on.

One known example: Pyruvic-Lactate ratio can be addressed by reducing the amount of lactic acid (i.e. no Lactobacillus probiotics, reducing food that produce lactic acid – milk, yogurt). It may also require 16s microbiome tests to determine which lactate producing bacteria are present (list here)

This is a continuing series of blogs looking at OATS results and autism. The goal is to filter out items that are relevant to autism. The series is not intended to do a full explanation of the OATS test. The OATS test is not autism specific but general health (often IBS/FM/CFS focuses).

Remember that our knowledge is constantly changing (unfortunately most MDs knowledge of the literature is stale).

Panel A

Glutathione

In this test panel, we see that this it is well below the normal range.

the functional properties (such as galactose metabolism, glycosyltransferase activity, and glutathione metabolism) displayed significant differences between the ASD and HC groups. The current study provides evidence for the relationship between gut microbiota and ASD, with the findings suggesting that gut microbiota could contribute to symptomology.

Gut microbiota changes in patients with autism spectrum disorders [2020]

• The role of glutathione redox imbalance in autism spectrum disorder: A review [2020] “, the existing data provide a strong background on the role of the glutathione system in ASD pathogenesis. Future research is necessary to investigate the role of glutathione redox signaling in ASD, which could potentially also lead to promising therapeutics.”
• “N-acetylcysteine, which can be converted to glutathione” [2020] In prior post, NAC was found to help a subset.
• Does infectious fever relieve autistic behavior by releasing glutamine from skeletal muscles as provisional fuel? [2013]
  • ” If glutamine released by fever rarely aggravates autistic behavior, why would supplemental glutamine?” i.e. direct supplementation of glutamine is not suggested.

Methylation

In this test panel, we see that this it is well above the normal range. There is a lot of literature dealing with methylation, so I have selected two recent studies dealing with the DNA aspect.

• Phenotypic Subtyping and Re-Analysis of Existing Methylation Data from Autistic Probands in Simplex Families Reveal ASD Subtype-Associated Differentially Methylated Genes and Biological Functions [2020]
  • “We demonstrate that subphenotyping of cases enables the identification of over 1.6 times the number of statistically significant differentially methylated regions (DMR) and DMR-associated genes (DAGs) between cases and controls,” This has a DNA dimension
• “it has been recently highlighted that glutathione can affect and modulate DNA methylation and epigenetics. ” [2020] – see above

Ammonia

This person is low in Ammonia Excess. The literature suggests level may depend on subset:

• Dysregulated amino acid metabolism, high ammonia and oxidative stress were prevalent among autistic children and should be considered in autism management [2020]
•  Twenty-eight (38%) cases were positive for H. pylori antigen in their stool with significant higher serum ammonia and lower adenosine deaminase than in H. pylori-negative autistic children.  [2019]
• “when concentrations of fecal acetic, butyric, isobutyric, valeric, isovaleric and caproic acids were measured, all were significantly higher in children with ASD compared with controls except for caproic acid. The concentration of fecal ammonia was also significantly greater in ASD participants than controls “[2012]

Aspartame, Salicylates and 2-Hydroxyhippuric returned no results.

Panel B

Summary of searches on PubMed for Autism with:

• 2-Hydroxyisovaleric – nothing
• 2-Oxoisovaleric – nothing
• 3-Methyl-2-oxovaleric – nothing
• 2-Hydroxyisocaproic – nothing
• 2-Oxoisocaproic – nothing
• 2-Oxo-4-methiolbutyric (Nothing found on this chemical on PubMed!!)
• Mandelic – nothing
• Phenyllactic and Phenylpyruvic – one research article on rats
  • “potentially explaining the origin of trans-indolylacryloylglycine, a postulated marker for autism.” [2012]
  • A low level could suggest “differences in the gut microbially‐mediated metabolism of phenylalanine”
• Homogentisic – nothing
• 4-Hydroxyphenyllactic – nothing
• N-Acetylaspartic – nothing
• Malonic – high levels seen at birth has increased risk of autism [2017]
• 4-Hydroxybutyric – nothing

Phosphoric has some 43 search hits for phosphorus. The most significant items are below

• “There was a significant correlation of levels of phosphorus and sulfur in the children with ASD” [2020] Higher –> Autism
• Several articles cited the importance of phosphorus for Vitamin D

Section Summary

None of the lab’s measurement here are relevant to autism. They may be relevant to other health issue.

Bottom Line

We see that DNA is involved with the possibility of altering its behavior with supplements and/or microbiome alteration.

The last of the test results forwarded by a reader was the Great Plains Laboratory Organic Acid Test. The two earlier tests results with comments are:

• Parasitology and Autism
• Minerals and Autism

There are a lot of tables in this report and I will do just 1-2 in each blog post to keep information flow manageable.

This test is interesting because it lists some of the bacteria associated associated various markers.

The one interesting thing is that most of the lows are associated to Aspergillus. This agrees with a study that I cited earlier:

“Saccharomyces and Aspergillus showed significant differences between ASD (59.07%) and Control (40.36%), indicating that they may be involved in the abnormal gut fungal community structure of ASD. When analyzed at the species level, a decreased abundance in Aspergillus versicolor was observed while Saccharomyces cerevisiae was increased in children with ASD relative to controls. ” [2020]

 Aspergillus oryzae is the only Aspergillus that I know that is available as a probiotic (Strong Wakamoto W)

 2-Hydroxyphenylacetic Acid is shown as high above. The literature mentions 3-Hydroxyphenylacetic Acid is common with Autism

• Increased urinary excretion of a 3-(3-hydroxyphenyl)-3-hydroxypropionic acid (HPHPA), an abnormal phenylalanine metabolite of Clostridia spp. in the gastrointestinal tract, in urine samples from patients with autism and schizophrenia [2010]
• Investigation of the relation between anaerobic bacteria genus clostridium and late-onset autism etiology in children [2014]
• Urinary 3-(3-Hydroxyphenyl)-3-hydroxypropionic Acid, 3-Hydroxyphenylacetic Acid, and 3-Hydroxyhippuric Acid Are Elevated in Children with Autism Spectrum Disorders [2016]
  • “After oral vancomycin treatment, urinary excretion of HPHPA (p < 0.001), 3HPA (p < 0.005), and 3HHA (p < 0.001) decreased markedly, which indicated that these compounds may also be from gut Clostridium species. … Additionally, the sensitivity and specificity data assessed by ROC analysis demonstrate that the measurements of the three metabolites are strong indicators of ASDs.”
  • ” after two therapeutic course treatments, the ABC score decreased significantly (mean value from 73 to 59); 90% autistic children showed improved communication and eye contact, but no obvious improvement in stereotyped behavior was seen. “

Nutritional Markers

The item that stands out is thiamin and there is no literature connecting it to autism beyond lower intake by diet in some locations.

• Autism spectrum disorder group failed to meet dietary recommendations for thiamin, riboflavin, vitamin C, or calcium. [2017]

From this article Nutritional and metabolic status of children with autism vs. neurotypical children, and the association with autism severity [2011] we read:

• No statistical difference for Thiamin
• No statistical difference for Riboflavin
• No statistical difference for Niacin
• No statistical difference for Pantothenic Acid, but average was lower
• No statistical difference for Vitamin C

I should add one word of warning, because the levels were similar, it does no exclude supplementation as being beneficial in some cases. The reason is that often they are processed into other chemicals (depending on available surplus and the bacteria present).

N-acetylcysteine (NAC) is more interesting because the marker indicate it was high. For a subset of ASD patients (high irritability), supplementation helps:

• The Use of N-acetylcysteine Supplementation to Decrease Irritability in Four Youths With Autism Spectrum Disorders [2020]
• A randomized double blind placebo controlled clinical trial of N-Acetylcysteine added to risperidone for treating autistic disorders [2013]
  • “Risperidone plus NAC more than risperidone plus placebo decreased irritability in children and adolescents with ASD.”
• A randomized placebo-controlled pilot study of N-acetylcysteine in youth with autism spectrum disorder [2016]
  • “The results of this trial indicate that NAC treatment was well tolerated, had the expected effect of boosting GSH production, but had no significant impact on social impairment in youth with ASD.“

CoQ 10 is shown as low above. As with NAC, supplementation appears to benefit a subset.

• Coenzyme Q ₁₀ supplementation reduces oxidative stress and decreases antioxidant enzyme activity in children with autism spectrum disorders [2018]” 
  • high doses ( daily doses of 30 and 60 mg. ) of CoQ₁₀ can improve gastrointestinal problems (P = 0.004) and sleep disorders (P = 0.005) in children with ASDs“
• Ubiquinol improves symptoms in children with autism [2014]
  • Ubiquinol supportive therapy improved symptoms in children with autism, as communication with parents (in 12%), verbal communication (in 21%), playing games of children (in 42%), sleeping (in 34%), and food rejection (in 17%), 

Bottom Line

My working model is that symptoms are strongly associated to the metabolites (chemicals/vitamins) produced/processed by the microbiome (bacteria in digestive system). For CoQ10 and NAC, we know the bacteria that are likely impacted, and that information is linked to here (CoQ10 , NAC).

This means that it is not a one-size fit all approach to autism, but very individual – either based on symptoms OR on the microbiome. I am biased towards using the microbiome (16s reports from Biome Sight [“MICRO” as discount code], followed by Thryve Inside)

The same reader also included Doctor’s Data Comprehensive Stool Analysis / Parasitology x3. The results are below, and what I would expect to be common: Nothing detected. Toxoplasmosis plays an important role as a risk factor for autism, but not once autism is established [2020].

The test also reported on possible high Saccharomyces boulardii/cerevisiae, Klebsiella pneumoniae, and Proteus mirabilis.

• “Saccharomyces and Aspergillus showed significant differences between ASD (59.07%) and Control (40.36%), indicating that they may be involved in the abnormal gut fungal community structure of ASD. When analyzed at the species level, a decreased abundance in Aspergillus versicolor was observed while Saccharomyces cerevisiae was increased in children with ASD relative to controls. ” [2020]
  •  Aspergillus oryzae is the only Aspergillus that I know that is available as a probiotic (Strong Wakamoto W)
• Neither klebsiella pneumoniae nor Proteus mirabilis are reported in the literature as being significant for autism.

Declaration: I have little trust in the cfu/gram for detecting microbiome dysfunction. It is old school.

Bottom Line

Using this test for a child with autism is unlikely to produce productive results for most cases of autism. A 16s microbiome test (for example  Biome Sight with “MICRO” as discount code or Thryve Inside) would produce better information from newer technology at significantly less cost.

A reader forwarded some lab reports and ask for comments. The reader gracefully gave permission to share some data to aid in the discussion and that others may benefit.

Connection to Microbiome

The absence or surplus of minerals impact both bacteria growth and the metabolites produced by the bacteria. An example study from 45 years ago illustrates this.

The effect of low levels of strontium, boron, lithium, molybdenum, and fluorine, alone and in combination, on hydroxyapatite solubility, bacterial growth, and acid production in five antigenic types of Streptococcus mutans was investigated…. The results show that low levels of strontium and fluorine can significantly reduce…

Effect of trace elements on dissolution of hydroxyapatite by cariogenic streptococci 1975

Antibiotic use is known to almost completely inhibit excretion of mercury in rats due to alteration of gut flora. [2019]

“During a pilot experiment, we found that germ-free mice [mice specially bred to have no bacteria anywhere in their systems] were resistant to anemia,” says Shah, senior author on the paper. “The easiest explanation is that you’ve gotten rid of a trillion bacteria and they no longer need iron. But interestingly, we saw that the iron absorptive mechanisms were all highly upregulated in the absence of microbiota.”

Gut Microbiome Puts the Brakes on Iron Absorption [2020]

Mineral Abnormalities in Autism

The microbiome influences the absorption and elimination of minerals. The minerals impacts the microbiome. In short, the microbiome will bias the mineral mixture in favor of what it prefers (“biome-forming”). In some cases, the environment that a person lives in may also cause shifts in mineral contents (i.e. water high in some mineral, living in a polluted environment). DNA may come in as a further compounding factor. Let us see what is reported in the literature:

Eating Habits

Often parents will shift diet due to beliefs or due to a child preference/resistance.

• Children with autistic disorder showed low dietary intake of some micronutrients; calcium (Ca), magnesium (Mg), iron (Fe), selenium (Se) and sodium (Na), also they had significantly high intake of potassium (K) and vitamin C compared to healthy controls [2017]
• Children with autism spectrum disorder consume less protein, calcium, selenium, vitamin D, thiamine, riboflavin and vitamin B12 and more polyunsaturated fat acid and vitamin E than controls. [2019]
• These children consumed significantly fewer macronutrients compared with the children without ASD. In addition, the children with ASD had the highest rate of vitamin A deficiency,  [2016]
• Relative to controls, ASD children consumed fewer number of food items, particularly fruits, vegetables, and proteins; had significantly lower daily intake of potassium, copper and folate. [2017]
• ASD patients consumed in average more calories than controls (though with a high patient’s frequency above and below calorie range references), had a limited food repertoire, high prevalence of children with inadequate calcium, sodium, iron, vitamin B5, folate, and vitamin C intake. [2016]
• Nutrients least likely to be consumed in recommended amounts were vitamin A, vitamin E, fiber, and calcium. Children with ASD were more likely to consume vitamin/mineral supplements than typically developing children. Compared with parents of typically developing children, parents of children with ASD were more likely to report that their children were picky eaters and resisted trying new foods [2008]
• The results obtained showed that the intake of carbohydrates and slightly lower intakes of protein, fat, calcium, magnesium, phosphorus, iron, zinc, retinol, vitamin B2, vitamin B12, folic acid, and pantothenic acid were higher among children and adolescents with ASD than among those without ASD.  [2020]
• Risk for specific inadequacies included vitamin D (97% of the sample), fiber (91%) vitamin E (83%), and calcium (71%). Children with five or more nutritional inadequacies (n=55) were more likely to make negative statements during meals (P<0.05). [2018]
• However, high fibre intake was connected with a decreased α-diversity only in children with ASD. High carbohydrate and fibre intake influenced β-diversity, changing the abundance of Bacteroides and other genera, many of them members of the Clostidiaceae. Modulating food habits of ASD children can influence their gut microbiota composition. [2020]

Child food-resistance can be a challenge to normalizing food intake (which may result in symptom improvement because of it’s impact on the microbiome).

Hair Concentrations

The above chart used hair analysis. There is significant literature in this area:

• By comparing hair concentration of autistic vs nonautistic children, elevated hair concentrations were noted for aluminum, arsenic, cadmium, mercury, antimony, nickel, lead, and vanadium. Hair levels of calcium, iron, iodine, magnesium, manganese, molybdenum, zinc, and selenium were considered deficient. [2012]
• . The mean Levels of mercury, lead, and aluminum in hair of the autistic patients were significantly higher than controls. Mercury, lead, and aluminum levels were positively correlated with maternal fish consumptions, living nearby gasoline stations, and the usage of aluminum pans, respectively. [2015]
•  The children with autism had significantly (p<0.001) higher in-hair concentration levels of lead, mercury and uranium. There was no significant difference between the two groups in the other five toxic elements.[2005]
• Mean Calcium level in the hair of the case group was lower than the mean level of this element in the control group. Mean Arsenic and Lead concentration in the hair of children with ASD was statistically significantly higher than the mean concentration of this element in the hair of children without neurological disorders. [2020]
•  We have also identified factors associated with concentrations of Lead in blood of children with ASD or suspected of having ASD, including dietary factors. These factors include child’s sex, parental education, exhibiting pica, and eating watermelon, lamb, and cold breakfast such as cereal. [2019]
• Children with autism had significantly (2.1-fold) higher levels of mercury but similar levels of lead and similar levels of zinc. Children with autism also had significantly higher usage of oral antibiotics during their first 12 mo of life, and possibly higher usage of oral antibiotics during their first 36 mo of life. [2007]
• The significant elevation in the concentration of Copper, Lead, and Mercury and significant decrease in the concentration of Magnesium and Selenium observed in the hair and nail samples of autistic subjects could be well correlated with their degrees of severity. [2011]

Risks from Minerals

•  our findings indicated that among children with the Ile/Ile genotype, those with blood manganese concentrations > 12 μg/L had about 4 times higher odds of ASD compared to those with blood manganese concentrations <12 μg/L. [2019]
• In the autistic groups, decreased concentration of protein in both hair and nail samples was observed…..Lower protein content and higher percentage of nitration in hair and nail of autistic children correlated with their degrees of severity. [2011]
•  Increasing hair Mercury concentrations significantly correlated with increased ASD severity [2012]
• In this study, it is evident that levels of mercury and copper in hair are significantly associated with higher Childhood Autism Rating Scale scores. This was supported by Adams et al. [19] who found that severity of a child’s autism coincided with the levels of toxic metals excreted in their urine after treatment with metal removal therapy; the higher the levels of antimony and other metals excreted, the more severe was the child’s autism.[2011]

Treatment

• Treatments that could address redox metabolism abnormalities include methylcobalamin with and without folinic acid in open-label studies and vitamin C and N-acetyl-l-cysteine in DBPC studies. These studies have reported improved core and associated ASD symptoms with these treatments. [2014]
• Dietary supplements, especially multivitamin/minerals, were used by 56% of children with ASD. The most common micronutrient deficits were not corrected (vitamin D, calcium, potassium, pantothenic acid, and choline) by supplements. Almost one-third of children remained deficient for vitamin D and up to 54% for calcium. Children receiving GFCF diets had similar micronutrient intake but were more likely to use supplements (78% vs 56%; P=0.01). Supplementation led to excess vitamin A, folate, and zinc intake across the sample, vitamin C, and copper among children aged 2 to 3 years, and manganese and copper for children aged 4 to 8 years. [2015]

Looking at the Results

• Zirconium [LOW]- no literature that could be applied on its impact
• Lithium [LOW]– supplementation should be discussed with your MD.
  • Lithium as a rescue therapy for regression and catatonia features in two SHANK3 patients with autism spectrum disorder: case reports [2015]
  • Lithium ameliorates autistic-like behaviors induced by neonatal isolation in rats [2014]
  • A new look at an old drug: neuroprotective effects and therapeutic potentials of lithium salts [2016]
• Antimony [HIGH] – It impacts several bacteria [2012]
  • We demonstrate that fluoride and aluminum (Al³⁺) can exacerbate the pathological problems by worsening excitotoxicity and inflammation.[2018]
• Mercury [HIGH] – well associated to autism and severity.
• Aluminum [HIGH] – well associated to autism
• Selenium [LOW]  – it is significant for antioxidants.

Selenium (₃₄Se), an antioxidant trace element, is an important regulator of brain function. These beneficial properties that Se possesses are attributed to its ability to be incorporated into selenoproteins as an amino acid. Several selenoproteins are expressed in the brain, in which some of them, e.g. glutathione peroxidases (GPxs), thioredoxin reductases (TrxRs) or selenoprotein P (SelP), are strongly involved in antioxidant defence and in maintaining intercellular reducing conditions. Since increased oxidative stress has been implicated in neurological disorders, including Parkinson’s disease, Alzheimer’s disease, stroke, epilepsy and others, a growing body of evidence suggests that Se depletion followed by decreased activity of Se-dependent enzymes may be important factors connected with those pathologies. 

Selenium in the Therapy of Neurological Diseases. Where is it Going? 2016

Bottom Line

While the following common is common with Autism, it does not apply to every child. Individual testing is strongly recommended.

• High Levels of:
  • Aluminum,
  • Antimony,
  • Arsenic,
  • Cadmium,
  • Lead,
  • Mercury,
  • Nickel,
  • Vanadium
• Low Levels of:
  • Boron,
  • Calcium,
  • Fluorine,
  • Iodine,
  • Iron,
  • Lithium,
  • Magnesium,
  • Manganese,
  • Molybdenum,
  • Selenium,
  • Strontium,
  • Zinc

Actions?

From prior readings, I know that items like Vitamin D absorption can be greatly influence by the microbiome. People take recommended dosages (which works across a typical population) and there is no significant changes. One approach is to use mega dosages. Another is to encourage bacteria known to assist absorption. For vitamin D, we know that Bifidobacterium Longum, Lactobacillus Casei and Lactobacillus Reuteri plays a role. Our knowledge is still fragmentary.

We have information on the bacteria influenced by Iron, dietary protein and a few more.

For items like aluminum being high, we have the association with aluminum cookware from the literature. We should be wary of all canned drinks because the container is aluminum and the contents are usually acidic (acid is used to extract aluminum). This also applies to items in TetraPaks(5% aluminum) which may include milk, soups.

Bottle waters are often no better (and sometimes worst) than tap water (2019 Consumer Reports Article).

For items like Selenium, always having a few Brazil Nuts around should address this issue. Using crude sea salt provides fluoride and iodine. A hot bed-time drink made with Cocoa powder(NOT chocolate favoring) helps with magnesium and zinc. Vanadium can be a bit of a challenge to get some kids to eat radishes, unlike molybdenum that may be obtained from potatoes.

There was a number of B-vitamins reported deficient cited above. I often refer to the B-vitamins as the Beef-Vitamins. The low protein intake would be expected to result in low levels of these. My first goal would be to increase protein intake before supplementation.

My future research

This has gotten me interested in the minerals-bacteria interaction. An area that I expect sparse information. As I find information I will post here.

FMTs have been tried for various conditions with mixed success. Autism has distinctive microbiome shifts and thus FMT should be consider as a treatment option. The why of failures has been an ongoing interest of mine. We may now have a significant factor that has been ignored in these attempts.

 Fecal microbiota transplantation (FMT) as a special organ transplant therapy, which can rebuild the intestinal flora, has raised the clinical concerns. It has been used in the refractory Clostridium difficile, inflammatory bowel disease, irritable bowel syndrome, chronic fatigue syndrome, and some non-intestinal diseases related to the metabolic disorders. But this method of treatment has not become a normal treatment, and many clinicians and patients can not accept it. 

[Research progress of fecal microbiota transplantation] 2015

This week’s Economist had an extended essay on Viruses and the like: The aliens among us/The Outsider within, this provides good background.

In addition to this, there was a podcast reporting success with FMT was associated with higher Phage Diversity in the donor. Phages are the police of the microbiome.

In this retrospective analysis, FMTs with increased bacteriophage α-diversity were more likely to successfully treat rCDI. In addition, the relative number of bacteriophage reads was lower in donations leading to a successful FMT. These results suggest that bacteriophage abundance may have some role in determining the relative success of FMT.

The success of fecal microbial transplantation in Clostridium difficile infection correlates with bacteriophage relative abundance in the donor: a retrospective cohort study (2019)

My earlier posts on FMT

• Fecal Microbiota Transplantation (FMT)
• An ideal FMT Donor scenario for ME/CFS
• Fecal Transplants – not for the herx adverse!
• A Healthy FMT Donor Microbiome and response to FMT
• uBiomes before and after a Fecal Microbiota Transplant

Bottom Line

This implies that for a greater chance of success and less risk, than DYI fecal transfer, that a lab that tests for possible infections AND for phage state may yield the best results.

I just pushed an update to the Special Condition Modifier page. You will see two links to suggestions on the page. See the earlier post here.

Pub Med

Citizen Science

Convergence!

There is a considerable one to one agreement with the suggestions. There is also agreement with gluten-free diet (i.e. no wheat, barley) which had positive results reported from studies (as well as B6).

In short, the microbiome analysis with suggestions appear to match actual studies — except we have more items suggested than been studied independently.

Implication for Specific Children

Above we are working from aggregations of many children with ASD. What is reported is not specific to one child. With an individual 16s stool sample test, then we can process that child’s unique individual microbiome profile thru and get suggestions specific for that child using the same logic as shown above.

The following 16s Providers provide data in an uploadable format. Biome Sight with “MICRO” as discount code and Thryve Alive

This is usually called hypoperfusion ( Hypoperfusion is a term that describes “a reduced amount of blood flow”. ) and is seen in SPECT scans of the brain. I am experienced with it from episodes of ME/CFS relapse (SPECT scan reports) that had poor memory, poor decision making, easy mental fatigue, increased irritability during the relapse. It is my belief that metabolic changes induced by microbiome dysfunction was the cause of hypoperfusion.

There are some interesting similarities between CFS/ME and ASD, for example:

• CFS/ME has increased Gray Brain Matter and decreased White Brain matter [2017]
• ASD Has increased Gray Brain Matter [2006]

Literature on Autism and hypoperfusion

• Cerebral Perfusion Abnormalities in Children With Autism and Mental Retardation: A Segmental Quantitative SPECT Study [2009] “Generalized hypoperfusion of brain was observed in all 10 cases as compared to controls. Frontal and prefrontal regions revealed maximum hypoperfusion. Subcortical areas also indicated hypoperfusion. We conclude that children with autism have varying levels of perfusion abnormities in brain causing neurophysiologic dysfunction that presents with cognitive and neuropsychological defects.”
• Technetium-99m HMPAO Brain SPECT in Autistic Children and Their Families [2008] ” In parents of AC, significant hypoperfusion was noted in the right parietal and bilateral inferior frontal cortex. In siblings of AC, perfusion in the right frontal cortex, right nucleus caudate and left parietal cortex was significantly decreased. “
• Regional Cerebral Blood Flow in Childhood Autism: A SPET Study With SPM Evaluation [2008] ” abnormal areas are related to the cognitive impairment observed in autistic children, such as language deficits, impairment of cognitive development and object representation, and abnormal perception and responses to sensory stimuli. “
• Effects of route of administration on oxytocin-induced changes in regional cerebral blood flow in humans [2020]
  • see Autism and Oxytocin levels, low levels are seen with ASD
• Arterial Spin Labeling Provides a Reliable Neurobiological Marker of Autism Spectrum Disorder [2018] “Arterial spin labeling revealed hypoperfusion in children with ASD in regions critical to social perception and cognition. “
• Altered Resting Perfusion and Functional Connectivity of Default Mode Network in Youth With Autism Spectrum Disorder [2015] “A pattern of altered resting perfusion was found in ASD versus TD children including frontotemporal hyperperfusion and hypoperfusion in the dorsal anterior cingulate cortex. “
• Hyperbaric Oxygen Treatment in Autism Spectrum Disorders [2012] ” A number of individuals with ASD possess certain physiological abnormalities that HBOT might ameliorate, including cerebral hypoperfusion, inflammation, mitochondrial dysfunction and oxidative stress….  A number of individuals with ASD possess certain physiological abnormalities that HBOT might ameliorate, including cerebral hypoperfusion, inflammation, mitochondrial dysfunction and oxidative stress…Studies which used a higher frequency of HBOT sessions (e.g., 10 sessions per week as opposed to 5 sessions per week) generally reported more significant improvements.”
• Brain Perfusion SPECT and EEG Findings in Children With Autism Spectrum Disorders and Medically Intractable Epilepsy [2010] ” In all children, eZIS revealed a mixed hypoperfusion pattern, especially in the prefrontal cortex, medial frontal cortex, anterior cingulate cortex, medial parietal cortex, and/or anterior temporal cortex.”
• Executive Function Deficits and Neural Discordance in Children With Autism Spectrum Disorders [2009] “Children with ASD were impaired in everyday executive functioning and response inhibition. The cordance value, which has been shown to correlate with brain perfusion in a number of studies, was significantly correlated with executive dysfunctions.”
• Brain Perfusion in Autism Varies With Age [2002] “As the age of the autistic individuals increased the hypoperfusion of verbal-associated areas in the left temporal lobe and frontal areas became more evident. The findings were significant at the p < 0.001 level. The changes in perfusion over time correlated with language development and acquisition as individuals matured. “
• The Relationship Between 99mTc-HMPAO Brain SPECT and the Scores of Real Life Rating Scale in Autistic Children [2002] “There was a relationship between bilateral F regions perfusion on 99mTc-HMPAO brain SPECT and the age of autistic children. There was also a negative correlation between IQ levels and the scores of sensory responses, social relationship to people, and sensory-motor responses”
• Perfusion Impairments in Infantile Autism on technetium-99m Ethyl Cysteinate Dimer Brain Single-Photon Emission Tomography: Comparison With Findings on Magnetic Resonance Imaging [1999] ” In conclusion, extensive perfusion impairments involving the cerebellum, thalami and parietal cortex were found in this study. SPECT may be more sensitive in reflecting the pathophysiology of autism than MRI.”
  • Personal note: With ME/CFS SPECT was also found to be more sensitive than MRI. My MRI was normal, my SPECT was not.

Treating Hypoperfusion

For ME/CFS, my treatment included sublingual heparin, piracetam and a variety of other items. Below are studies on various items that impacts hypoperfusion.

• Piracetam Ameliorated Oxygen and Glucose Deprivation-Induced Injury in Rat Cortical Neurons via Inhibition of Oxidative Stress, Excitatory Amino Acids Release and P53/Bax [2014]
  • Piracetam Improves Cognitive Deficits Caused by Chronic Cerebral Hypoperfusion in Rats [2008]
• Supplementary Management With Pycnogenol® in Patients With Lupus Vasculitis in Remission Phases: A Pilot, Concept Registry Study [2020]
• Intake of Psyllium Seed Husk Reduces White Matter Damage in a Rat Model of Chronic Cerebral Hypoperfusion [2019]
• Resveratrol Reverses the Synaptic Plasticity Deficits in a Chronic Cerebral Hypoperfusion Rat Model [2016] a.k.a. Grape Seed Extract
  • Chronic Treatment With Resveratrol, a Natural Polyphenol Found in Grapes, Alleviates Oxidative Stress and Apoptotic Cell Death in Ovariectomized Female Rats Subjected to Chronic Cerebral Hypoperfusion [2016]
• Role of Vinpocetine in Cerebrovascular Diseases [2011]
• Effects of Coenzyme Q and Creatine Supplementation on Brain Energy Metabolism in Rats Exposed to Chronic Cerebral Hypoperfusion [2011]
• Eicosapentaenoic Acid: Effect on Brain Prostaglandins, Cerebral Blood Flow and Edema in Ischemic Gerbils [1984] ” Our study indicates that eicosapentaenoic acid prevented post-ischemic cerebral edema and hypoperfusion, without affecting the levels of brain diene prostaglandin and thromboxane.” An Omega-3 acid.
• L-Arginine and Superoxide Dismutase Prevent or Reverse Cerebral Hypoperfusion after Fluid-Percussion Traumatic Brain Injury [2009] only L-arginine was effective

Possible Prescription Drugs

• Alleviation of Brain Hypoperfusion after Preventative Treatment with Lomerizine in an Elderly Migraineur with Aura [2011] (calcium channel blocker)
• [Heparin–piracetam complex and its effect on blood and blood circulation].1998 ““The heparin–piracetam complex had a more pronounced anticoagulant and fibrinolytic activities than the individual components. In addition, this complex accelerated blood flow in the brain.”

Low level coagulation as a contributor to hypoperfusion

For myself with ME/CFS, activation of coagulation was a significant factor and confirmed by labs tests from Hemex (this battery of tests is still available from one lab). Blood flow to the brain can be caused by:

1. ‘thick blood'(think of a heavy oil(molasses) versus a light oil (water), one moves much slower than the other). Since blood delivers oxygen, it means less oxygen
2. fibrin fibers (‘dirty filters’ that slows the slow, a blood clot would stop the flow)

What do we know from the literature on coagulation and autism?

• Autism, an Extreme Challenge to Integrative Medicine. Part: 1: The Knowledge Base [2002] “Coagulation abnormalities have been reported.”
• Platelet studies in autism spectrum disorder patients and first-degree relatives [2015] “We report increased platelet counts, decreased platelet ATP dense granule secretion, and increased serotonin plasma levels not only in ASD patients but also in their first-degree relatives. This suggests that potential genetic factors associated with platelet counts and granule secretion can be associated with, but are not fully penetrant for ASD.”
• Link Between Autism And Abnormal Blood-vessel Function And Oxidative Stress [2006] “children with autism showed signs of abnormal blood-vessel function and damaging levels of oxidative stress compared to healthy children. The children with autism possessed levels of biochemicals that indicate the presence of constricted blood vessels via the endothelium (the cells that line vessels) with a higher tendency to form clots (through cells called platelets). “
• Evaluation of Platelet Parameters in Children with Autism Spectrum Disorder: Elongated Collagen‐Adenosine Diphosphate and Collagen‐Epinephrine Closure Times [2019]

Most of these issues are replicated in findings with ME/CFS (see links on this page: https://me-pedia.org/wiki/David_Berg).

There are many dimensions here, researched items that I have used include:

• Alpha Lipoic Acid
• Lumbrokinease
• Nattokinease
• Bromelain
• Turmeric

Bottom Line

In doing this post it was a bit of a surprise to see that 1st degree relatives was seen with similar conditions. For myself, it was not because I have an inherited coagulation defect (Prothrombin G20210A a.k.a. Factor II Mutation) so 1st degree relatives having it is to be expected.

The role of the microbiome and diet for hypoperfusion is not well explore. Emerging Role of Diet and Microbiota Interactions in Neuroinflammation [2018] gives an overview, but implications for hypoperfusion in autism is a to be determined.

This is an EDUCATIONAL POST, the items discussed above (including supplements) should be discussed with your medical professional before starting. This is not medical advice.