“Why Most Published Research Findings Are False” or “Much of the scientific literature, perhaps half, may simply be untrue”

Regular readers of this blog, and the small number of other autism science blogs, will have noticed how often research findings are contradictory.  In autism there is often a good reason why things are contradictory, because in some of the underlying dysfunctions both too much, or too little, (hyper or hypo) both lead to “autism”.     

Take the relatively simple case of GI problems in autism; estimates of prevalence range from 9 to 91%.  Such a variation is of course absurd and on the basis of which study you choose to quote, you can make whatever case you want.

   

Given that autism is not a single disease like say rheumatic fever, another very odd inflammatory disease, it makes anything very much harder to prove.

In the wider world of medical research there is an ongoing debate about research studies in general.  Most of this research is in much simpler types of disease, where you can accurately include in your trial only people with the same single biological dysfunction.

It has been suggested recently in the Lancet, one of the world’s top medical journals, that perhaps as much of half of the findings in research are untrue.

Much of the scientific literature, perhaps half, may simply be untrue.

   

The case against science is straightforward: much of the scientific literature, perhaps half, may simply be untrue. Afflicted by studies with small sample sizes, tiny effects, invalid exploratory analyses, and flagrant conflicts of interest, together with an obsession for pursuing fashionable trends of dubious importance, science has taken a turn towards darkness. As one participant put it, “poor methods get results”. The Academy of Medical Sciences, Medical Research Council, and Biotechnology and Biological Sciences Research Council have now put their reputational weight behind an investigation into these questionable research practices. The apparent endemicity of bad research behaviour is alarming. In their quest for telling a compelling story, scientists too often sculpt data to fit their preferred theory of the world. Or they retrofit hypotheses to fit their data. Journal editors deserve their fair share of criticism too. We aid and abet the worst behaviours. Our acquiescence to the impact factor fuels an unhealthy competition to win a place in a select few journals. Our love of “significance” pollutes the literature with many a statistical fairy-tale. We reject important confirmations. Journals are not the only miscreants. Universities are in a perpetual struggle for money and talent, endpoints that foster reductive metrics, such as high-impact publication. National assessment procedures, such as the Research Excellence Framework, incentivise bad practices. And individual scientists, including their most senior leaders, do little to alter a research culture that occasionally veers close to misconduct.

Can bad scientific practices be fixed? Part of the problem is that no-one is incentivised to be right. Instead, scientists are incentivised to be productive and innovative. Would a Hippocratic Oath for science help? Certainly don't add more layers of research red-tape. Instead of changing incentives, perhaps one could remove incentives altogether. Or insist on replicability statements in grant applications and research papers. Or emphasise collaboration, not competition. Or insist on preregistration of protocols. Or reward better pre and post publication peer review. Or improve research training and mentorship. Or implement the recommendations from our Series on increasing research value, published last year. One of the most convincing proposals came from outside the biomedical community. Tony Weidberg is a Professor of Particle Physics at Oxford. Following several high-profile errors, the particle physics community now invests great effort into intensive checking and re-checking of data prior to publication. By filtering results through independent working groups, physicists are encouraged to criticise. Good criticism is rewarded. The goal is a reliable result, and the incentives for scientists are aligned around this goal. Weidberg worried we set the bar for results in biomedicine far too low. In particle physics, significance is set at 5 sigma—a p value of 3 × 10^–7 or 1 in 3·5 million (if the result is not true, this is the probability that the data would have been as extreme as they are). The conclusion of the symposium was that something must be done. Indeed, all seemed to agree that it was within our power to do that something. But as to precisely what to do or how to do it, there were no firm answers. Those who have the power to act seem to think somebody else should act first. And every positive action (eg, funding well-powered replications) has a counterargument (science will become less creative). The good news is that science is beginning to take some of its worst failings very seriously. The bad news is that nobody is ready to take the first step to clean up the system.

Ten years ago John Ioannidis, a well-known Professor of Medicine now at Stanford, published a much more complex paper saying essentially the same thing.  That paper, below, was recently highlighted to me by one reader of this blog, who is involved in such research.

  

Why Most Published Research Findings Are False

Conclusion

  

Since modern medicine, and indeed some autism therapy, is all about being “evidence based”, this does pose a big problem.

When the experts are telling us not to trust the evidence, what/who do you trust?

Getting into debates about almost anything to do with autism, with anyone, usually gets you nowhere at all.  Even the most basic points are disputed. This is at least one thing that people can agree on.

In the end it comes down to your own experience and judgement; I hope it is good.

In spite of all its shortcomings, I continue to marvel at how easy it is to access what you want from the vast amount of accumulated research, draw your own conclusions and act on them. I wish more people did the same.

Time for a new generation of Autism “Experts”?

Farewell to Pingu

I usually succeed in keeping to the science in this blog and refrain from sharing my other wider opinions.  Today I slipped up.

Rather like James Simons, founder of the Simons Foundation, I do not have a high opinion of many supposed experts, particularly when it comes to autism.

Autism is often a backwater, where “academics” can still write the occasional quasi-scientific paper on some touchy-feely aspect that they consider important and make a name for themselves.  On the back of this, they can advocate for their perception of autism and often encourage ever-wider diagnosis in people who are less and less severely affected.

So far this is fine; we are all entitled to have our own opinions, so no comments on Autism Speaks, Autism One, or the various National Autism Societies. They all have the best intentions.

I should highlight the Simons Foundation and the UC Davis MIND Institute as being excellent scientific sources of objective information.

The only reason for today’s post is the comment made by someone who might be seen as the United Kingdom’s top autism expert, Professor Simon Baron-Cohen, Director of Cambridge University's Autism Research Centre. With such a tittle, he should set a high standard.

http://www.bbc.co.uk/programmes/p0312292

"We're not looking for a cure  … its part of their genetic and neurological make up," said Professor Simon Baron-Cohen, director of the Autism Research Centre at Cambridge University and vice-president of the National Autistic Society.

And …

“Children with Autism wait on average about three and a half years to get their diagnosis," Mr Baron-Cohen said.

It may come as a surprise to the Professor, but Parkinson’s disease is also a complex condition with a complex genetic element.  

Numerous very bright researchers are working to defeat Parkinson’s disease and, I am informed, that they are quite likely to achieve this end.

They set out to find a cure; they may indeed fall short and just find an effective therapy.

Imagine athletes starting a race, they all aim for gold.  By aiming for first place you might just come 5^th, but if you did not train for gold you would come last.

As for the Professor’s comment that people in the UK wait over three years for an autism diagnosis, this is not strictly true and is scaremongering.  People with what used to be autism, i.e. classic autism, Kanner’s autism or autistic disorder go to the front of the assessment queue and get diagnosed for free within a few months.  Some parents keep going back for a second, third, fourth, fifth opinion until they get the result they want; this takes years. 

There is of course nothing to stop someone in the UK paying themselves for the assessment and so waiting a matter of weeks; after all they would happily pay to have someone fix their washing machine.

The UK consultant neurodevelopmental pediatrician, who diagnosed my son, told us at the time that a multidisciplinary assessment is generally not needed and that she can see almost immediately if a 3-4 year old has autism.  With what I now know, I would have to agree with her 100%; to the trained eye it is obvious.  

There are numerous milder dysfunctions that can affect children and there is an ever growing raft of observational diagnoses.  These diagnoses are all highly subjective and some really could be simplified to “I DON’T KNOW”  (PDD NOS = Pervasive Developmental Disorder Not Otherwise Specified).  I rather liked IED = Intermittent Explosive Disorder.

In many cases the troubling symptom in these milder dysfunctions is “just” anxiety.  If the anxiety is severe enough to need treatment, why not try one of the safe (i.e. not Prozac) drugs? There are numerous existing treatments (Propranolol for example) for children with anxiety and some interesting new ones (Baclofen).

Then we would be left with:-

Autism, a disabling developmental disorder diagnosed in early childhood.  Following its diagnoses, based on simple observational/behavioral criteria, a biological diagnosis of the underlying dysfunctions should be sought.  A small minority of these dysfunctions are substantially curable.  Some dysfunctions in the majority of the remaining children are, to a meaningful extent, treatable today.

Having treated what is treatable, use behavioral therapy.

Unable to contribute to finding cures/therapies for the underlying biological dysfunctions, the Professor and many like him have sought to widen the scope of “autism” and diagnose ever more people.  The latest idea being the Broad(er) Autism Phenotype (BAP), where you diagnose almost anyone as a teeny tiny bit autistic.

Now everyone from Nikola Tesla to Fieldmarshal Montgomery is supposed to have been autistic.  Both did rather well for themselves for someone with a disability.

This “mild” autism is extremely subjective, which is why so many assessors will not diagnose it, and so yes, if you keep going for several years you may find someone to tell you that you have “autism”.  But then what?  There are no cures or therapies, apparently.  There are these supposed “services” which are available on diagnosis, depending where you live.

There are indeed valuable services for people with severe autism, like speech therapy, occupational therapy and behavioral therapy.  These are not going to help much with mild autism.

The mother of the child in the above BBC interview said her child is much better now because “he is on the right (anxiety) medication” and is in a school with small classes.  He had previously been given all sorts of diagnoses, Tourette syndrome, OCD etc. in multiple earlier assessments.

Why not take an anxiety medication if you had OCD, or Tourette’s?  Just google it.  Was the doctor only willing to prescribe an anxiety drug with an autism diagnosis?

The key was treating the anxiety and being in a small class at school.  Is that really “autism”?

Many children who are different, in one way or the other, struggle in large classes and many of them also get bullied, sometimes even by the teacher.  By all means call it Asperger’s, but please do not call it autism.

Yes, the Professor would say that autism is a spectrum.  This really means he has no clue what is the underlying biological dysfunction, so let’s call it all “autism” and be done with it.  “Autism” is in fact just a name for almost anything that goes mildly or severely wrong neurologically, in the first few years of life. 

Once you are a teenager, if a new unknown dysfunction occurs, it gets new labels: - schizophrenia, bipolar etc.  These are also just observational diagnoses and within them are numerous different genetic and environment causes, some of which are treatable, if you care to look.

Other Experts

It is not just the Professor; you will come across numerous “local experts” perpetuating misunderstandings regards autism.  Like the dedicated Principal of the autism special school explaining that some of her kids have such terrible anxiety that their stomachs are tied in knots and they have severe GI problems.  Anxiety certainly does not help, but we actually know much about reflux/GERD in autism and the more serious IBS/IBD/ulcerative colitis.  We even know the mechanism that may explain why reflux/GERD is comorbid with autism, it is called mGluR5.

On the subject of ulcerative colitis and autism, just because a certain Dr Wakefield highlighted this link, it is widely assumed to be a falsehood.  The literature is strewn with links between ASD and ulcerative colitis.  I have an anecdote of my own.  One of the few people I hear about with Asperger’s, where we live, was first diagnosed with ulcerative colitis, then followed the Asperger’s diagnosis, and now he needs part of his intestines removed.   This could have been treated years earlier and then there would be no need for surgery now and years of trouble at school could have been avoided.

Clever Scientists

What is needed are hard-core clever scientists, not soft-hearted touchy feely Psychologists. (Apologies to those forward looking Psychologists amongst you, who keep updated by reading the literature)

I could not agree more with James Simons and his Foundation, who choose to fund Nobel Laureates and future Nobel Laureates.  Hopefully, they do actually aim to find cures.  It may take a hundred years, but along the way there will be numerous therapies to improve outcomes in the meantime.

Some of these therapies for specific causes of “autism” already exist.  They are in the literature, but I guess Baron-Cohen does not read it.

Perhaps a little shocking is that even though the Professor is not a medical doctor and so has no medical experience of treating people with autism, he was the 2012 Chair of the National Institute for Health and Care Excellence (NICE) Guideline Development Group for adults with autism.  This is advice that is given to doctors in the UK on how to treat autism.  Not surprisingly, NICE guidelines to doctors in the UK actually tell them not to bother, core autism is not medically treatable.   

In many areas of health, like asthma, the NICE treatment guidelines are excellent and a great resource for clinicians and patients.

How can you attend the top universities, albeit not in medicine, work 25 years in  the field of autism research, travel to all those nice conferences, even edit an autism journal and not have realized/discovered that it is treatable? It is almost criminally negligent.  If cases of autism are treatable, they are potentially preventable and, if caught early enough, potentially reversible.   

If you want a warm feeling, don’t campaign for “Autism Awareness” or wider “Autism Diagnosis”, campaign for free detailed metabolic testing, genetic testing and MRIs (with MR spectroscopy) for all people diagnosed under five with autism (at their first assessment!).

By the way, I have no doubt that many of the highly intelligent researchers who get funding from the Simons Foundation would also have struggled in their childhood had they been in large classes, in non-selective, State-funded, primary schools.  

A wake up call

It is not just me claiming that most types of autism are, to varying degrees, treatable today.

There are a growing number of readers of this blog who have found the same. Most are regular parents, who current “Experts” would totally ignore, but some are actually doctors, medical researchers, and even Professors of Medicine.

If an amateur, with a blog, can figure out usable therapies from the literature, we really do need some new Experts, and then NICE will need to rewrite their guidelines.

Gene Silencers and Enhancers in Autism; plus Epicatechin, MOCOS, Ferritin and Oxidative Stress (GR, GPx, GCL, GCLM)

The original idea of this blog was to try to keep complicated things as simple as possible, so at times things may get over-simplified.  

This post starts out simple and then gets rather involved in oxidative stress.

When people think about genes, they are nearly always thinking about the “blueprints” that are encoded on your DNA.  As it turns out only about 5% of your DNA is dedicated to this function; this 5% is contained in the exome.

Much autism research is dedicated to finding faulty “blueprints” that might account for autism.  There are now several hundred so called “autism genes”, where an error in the “blueprints”, means that the associated protein is not produced to its intended specification.

We also have seen that genetic defects just lead to a possibility of something going wrong.  A “faulty gene” creates the possibility of a specific dysfunction happening, it does not mean 100% that it will happen. 

Partial dysfunctions and partial deficiency

We also saw that even when a single gene dysfunction, like for fragile-X, occurs it does not always cause a catastrophic failure, rather it produces a spectrum from mild to severe.

This point is important since it seems in autism there can often be “partial dysfunctions” leading to “partial deficiencies”.  This is just a less severe form of the “rare” total dysfunctions.  The growing list of examples includes partial biotinidase deficiency, partial glutathione reductase deficiency and partial glutathione peroxidase deficiency.  Today we will also encounter ferritin (iron storage) partial deficiencies.  In a future post we will look the vitamin B12 partial dysfunction that occurs in about a quarter of schizophrenia and autism cases.

This then leads us to the subject of gene expression, which means how much, where, when and how often a gene is turned “on”.  This is actually what really matters, since even perfectly good genes, when over-expressed, can do great damage.  We saw that in the case of Down Syndrome there is about 50% over expression in up to 300 genes.  In the case of Down Syndrome the reason for this overexpression lies in the exome.  In effect there is a double set of blueprints for those 300 genes.

Within the remaining 95% of your DNA are so-called enhancers and silencers.  Their job is to determine which genes are turned on (enhancers) or turned off (silencers) in which part of the body.  So a gene might encode a calcium channel, but that calcium channel should only be in certain parts of the body and only to a certain degree.  We need the correct clean blueprint and we need it applied in the right part of the body and only to the desired extent.

I was very pleased to see that some scientists have started to look at the role of enhancers in autism.  I have already noticed that some substances that are known to affect gene expression are particularly effective in autism.  This suggests to me that in some types of autism, the problem may actually be simply in gene expression rather than any faulty genetic “blueprint”.

Now the science of enhancers and particularly silencers is still at the emerging stage, but the research showed that in at least 100 locations, there were significant anomalies in those with autism.

Gene  function in the human brain could help reveal basis of autism

This is an easy to read summary of the research paper below.

Coexpression networks identify brain region–specific enhancer RNAs in the human brain

Abstract

Despite major progress in identifying enhancer regions on a genome-wide scale, the majority of available data are limited to model organisms and human transformed cell lines. We have identified a robust set of enhancer RNAs (eRNAs) expressed in the human brain and constructed networks assessing eRNA-gene coexpression interactions across human fetal brain and multiple adult brain regions. Our data identify brain region-specific eRNAs and show that enhancer regions expressing eRNAs are enriched for genetic variants associated with autism spectrum disorders.

We also have the removable markers on the 5% of DNA that cause epigenetic changes.  This is another way of turning on or off specific genes.  These markers can be caused by environment factors like smoking, or even stress, these markers are potentially both removable and inheritable.     

The emerging science of Proteomics is the study of gene expression itself, so it is measuring all the proteins that the genes actually produced.

Limits of Genetic Testing

So while in some cases genetic testing of the 5% of DNA usually examined may indeed be useful, if your problem was in the other 95% of DNA it will not help.

To be useful in autism you would need to measure gene expression in the brain or the local activity of the enhancers/silencers, since it varies throughout the body.  In the Australian study above they measured the enhancer activity in the brain, by looking for the special enhancer molecules the enhancers produce.

This is all way beyond the scope of this blog.

However when I see “safe” substances like Sulforaphane, Epicatechin and even statins that are known to affect the expression of multiple genes, I take note. 

Steroids also affect gene expression, but great care has to be taken with steroids.

Statins have numerous interesting effects in the brain and in cancer cells.  In autism they have an effect on PTEN and BCL2 for example.

Chronic Administration of Statins Alters Multiple Gene Expression Patterns in Mouse Cerebral Cortex

Statins down regulate myeloperoxidase gene expression in macrophages

Statins cause profound effects on gene expression in human cancer cellsin vitro: the role of membrane microdomains.

The observed impact of pravastatin on gene expression may explain the pleiotropic effects of statins when they are used as adjuvants in chemotherapy and suggests impact on gene expression as a possible cause of side effects from statin use.

As pointed out in the last paper, changing gene expression can be bad as well as good.  It all depends where you are starting from and what genes you want to enhance/silence.

Other therapies to modify gene expression

Today’s scientific knowledge does not always allow us to target the expression of specific genes, this very much remains future science.

However, the remarkable effects of some substances, in some people, does suggest some options.  As is often the case this takes us back to oxidative stress, which does seem to affect many conditions and is quite well studied. There is no shortage of anecdotal evidence.

We know from the research that oxidative stress is ever-present in autism and that people with autism are particularly sensitive to it.

One substance previously mentioned in this blog, epicatechin, is known to change the expression of many genes including STAT1, MAPKK1, MRP1, and FTH1, which are involved in the cellular response to oxidative stress.

Ferritin

Rather off subject the FTH1 gene encodes the heavy subunit of ferritin, the major intracellular iron storage protein.

Relationship of serum ferritin levels to sleep fragmentation and periodic limb movements of sleep on polysomnography in autism spectrum disorders.

Children with autism spectrum disorders had significantly lower ferritin levels compared with controls

Within the autism spectrum disorders population, median ferritin levels were significantly lower in patients with poor sleep efficiency (7 ng/mL) versus those with normal sleep efficiency (29 ng/mL) (P = 0.01).

Low ferritin would indicate an iron storage problem and likely anemia/anaemia. 

Low ferritin has many effects, including surprisingly, poor sleeping patterns.

  

Is it such a surprise that a cup of cocoa (epicatechin) before bed used to be given to ensure a good night’s sleep?  (all via FTH1, I presume)

Perhaps poor sleep in autism is just another consequence of oxidative stress?

MOCOS

In the recent paper on MOCOS:-

Olfactory stem cells reveal MOCOS as a new player in autism spectrum disorders

I noted that:-

Furthermore, we found that MOCOS misexpression induces increased oxidative-stress sensitivity.

MOlybdenum COfactor Sulfurase (MOCOS), is an enzyme involved in purine metabolism and a newly identified player in ASD. MOCOS appears to be downregulated in autism and this has multiple effects, one being increased sensitivity to oxidative stress.

Seemingly unknown to the French MOCOS researchers, there already is a therapy:-

Rescue of lethal molybdenum cofactor deficiency by a biosynthetic precursor from Escherichiacoli

Since I do not have any of the above biosynthetic precursor at hand, but I do have high flavanol cocoa in the kitchen, it is time to look again at epicatechin.

Epicatechin

There are two very similar substances catechin and epicatechin; both are flavonoids.  Both affect gene expression and both seem to have numerous good properties.

Epicatechin is found in large quantities in mildly processed cocoa, which catechin in found in large quantities in certain types of Chinese tea.

We saw in an earlier post that Mars, the chocolate company, has invested substantially in the science of cocoa and its flavonoids.  They have just signed a 5 year research contract with Harvard.

Catechin affects the fat metabolism and is therefore a potential therapy for obesity.  Oolong tea has been shown to have this effect, but you do need to drink a great deal of it.

Beneficial effects of oolong tea consumption on diet-induced overweight and obese subjects

CONCLUSIONS:

Oolong tea could decrease body fat content and reduce body weight through improving lipid metabolism. Chronic consumption of oolong tea may prevent against obesity.

  

The cocoa flavanol (-)-epicatechin protects the cortisol response

ABSTRACT Various health benefits of the cocoa flavanol (-)-epicatechin (EC) have been attributed to its antioxidant and anti-inflammatory potency. In the present study we investigated whether EC is able to prevent deterioration of the anti-inflammatory effect of the glucocorticoid (GC) cortisol in the presence of oxidative stress. It was found that cortisol reduces inflammation in differentiated monocytes. Oxidative stress extinguishes the anti-inflammatory effect of cortisol, leading to cortisol resistance. EC reduces intracellular oxidative stress as well as the development of cortisol resistance. This further deciphers the enigmatic mechanism of EC by which it exerts its anti-inflammatory and antioxidant action. The observed effect of the cocoa flavanol EC will especially be of relevance in pathophysiological conditions with increased oxidative stress and consequential GC resistance and provides a fundament for the rational use of dietary antioxidants

Flavanol-rich cocoa induces nitric-oxide-dependent vasodilation in healthy humans

  

Abstract

Background: Consumption of flavonoid-rich beverages, including tea and red wine, has been associated with a reduction in coronary events, but the physiological mechanism remains obscure. Cocoa can contain extraordinary concentrations of flavanols, a flavonoid subclass shown to activate nitric oxide synthase in vitro.

Objective: To test the hypothesis that flavanol-rich cocoa induces nitric-oxide-dependent vasodilation in humans.

Design: The study prospectively assessed the effects of Flavanol-rich cocoa, using both time and beverage controls. Participants were blinded to intervention; the endpoint was objective and blinded.

Methods: Pulse wave amplitude was measured on the finger in 27 healthy people with a volume-sensitive validated calibrated plethysmograph, before and after 5 days of consumption of Flavanol-rich cocoa [821 mg of flavanols/day, quantitated as (−)-epicatechin, (+)-catechin, and related procyanidin oligomers]. The specific nitric oxide synthase inhibitor, N^G-nitro-l-arginine methyl ester (l-NAME) was infused intravenously on day 1, before cocoa, and on day 5, after an acute ingestion of cocoa.

Results: Four days of flavanol-rich cocoa induced consistent and striking peripheral vasodilation (P = 0.009). On day 5, pulse wave amplitude exhibited a large additional acute response to cocoa (P = 0.01). l-NAME completely reversed this vasodilation (P = 0.004). In addition, intake of flavanol-rich cocoa augmented the vasodilator response to ischemia. Flavanol-poor cocoa induced much smaller responses (P = 0.005), and none was induced in the time-control study. Flavanol-rich cocoa also amplified the systemic pressor effects of l-NAME (P = 0.005).

Conclusion: In healthy humans, flavanol-rich cocoa induced vasodilation via activation of the nitric oxide system, providing a plausible mechanism for the protection that flavanol-rich foods induce against coronary events.

Cocoa Flavanols and Brain Perfusion

Flavanols, the Kuna, cocoa consumption, and nitric oxide

Abstract

The Kuna Indians, who reside in an archipelago on the Caribbean Coast of Panama, have very low blood pressure (BP) levels, live longer than other Panamanians, and have a reduced frequency of myocardial infarction, stroke, diabetes mellitus, and cancer—at least on their death certificates. One outstanding feature of their diet includes a very high intake of flavanol-rich cocoa. Flavonoids in cocoa activate nitric oxide synthesis in healthy humans. The possibility that the high flavanol intake protects the Kuna against high BP, ischemic heart disease, stroke, diabetes mellitus, and cancer is sufficiently intriguing and sufficiently important that large, randomized controlled clinical trials should be pursued.

Glutathione reductase (GR) and (partial) Glutathione reductase deficiency

Glutathione reductase (GR) catalyzes the reduction of glutathione disulfide (GSSG) to the sulfhydryl form glutathione (GSH), which is a critical molecule in resisting oxidative stress and maintaining the reducing environment of the cell.

Glutathione reductase reduces one mole of GSSG to two moles of GSH.

Glutathione reductase deficiency is a “rare” disorder in which the glutathione reductase activity is absent from erythrocytes, leukocytes or both. In one study this disorder was observed in only two cases in 15,000 tests for glutathione reductase deficiency performed over the course of 30 years. In the same study, glutathione reductase deficiency was associated with cataracts and favism in one patient and their family, and with severe unconjugated hyperbilirubinemia in another patient. It has been proposed that the glutathione redox system (of which glutathione reductase is apart) is almost exclusively responsible for the protecting of eye lens cells from hydrogen peroxide because these cells are deficient in catalase, the enzyme which catalyzes the breakdown of hydrogen peroxide, and the high rate of cataract incidence in glutathione reductase deficient individuals.

Some patients exhibit deficient levels of glutathione activity as a result of not consuming enough riboflavin in their diets. Riboflavin is a precursor for FAD, whose reduced form donates two electron to the disulfide bond which is present in the oxidized form of glutathione reductase in order to begin the enzyme's catalytic cycle.
In 1999, a study found that 17.8% of males and 22.4% of females examined in Saudi Arabia suffered from low glutathione reductase activity due to riboflavin deficiency.

Glutathione reductase deficiency in Saudi Arabia.

Abstract

Glutathione reductase (GR) is a ubiquitous enzyme required for the conversion of oxidized glutathione (GSSG) to reduced glutathione (GSH) concomitantly oxidizing reduced nicotinamide adenine dinucleotide phosphate (NADPH) in a reaction essential for the stability and integrity of red cells. Mutations in the GR gene and nutritional deficiency of riboflavin, a co-factor required for the normal functioning of GR, can cause GR deficiency. We conducted a study on 1691 Saudi individuals to determine the overall frequency of GR deficiency and to identify whether the deficiency results from genetic or acquired causes or both. The activity of GR was measured in freshly prepared red cell haemolysate in the presence and absence of flavin adenine dinucleotide (FAD) and the activity coefficient (AC) was determined. Samples with low GR activity (> 2.0 IU/g haemoglobin) both in the presence and absence of FAD and an AC between 0.9 and 1.2 were considered GR-deficient. Samples with AC > or = 1.3 were considered riboflavin-deficient. The overall frequency of partial GR deficiency was 24.5% and 20.3% in males and females respectively. In addition, 17.8% of males and 22.4% of females suffered from GR deficiency due to riboflavin deficiency. This could be easily corrected by dietary supplementation with riboflavin. No cases of severe GR deficiency were identified.

Regular readers may recall something very similar with biotin and its enzyme biotinidase.  Biotinidase deficiency is supposedly such a rare metabolic disorder that it is no longer screened for; however, in an autism study in Crete, Greece it was found that partial biotinidase deficiency was quite common.

Glutathione peroxidase

Glutathione peroxidase (GPx) is the general name of an enzyme family with peroxidase activity whose main biological role is to protect the organism from oxidative damage.

The biochemical function of glutathione peroxidase is to reduce lipid hydroperoxides to their corresponding alcohols and to reduce free hydrogen peroxide to water.

In earlier posts on anti-oxidants we saw the following presentation from the German scientist.  Note Glutathione (GSH) peroxidases, left halfway down

Glutamate Cysteine Ligase (GCL)

  

Glutamate Cysteine Ligase (GCL) is the first enzyme of the cellular glutathione (GSH) biosynthetic pathway.

GSH, and by extension GCL, is critical to cell survival.

Nearly every eukaryotic cell, from plants to yeast to humans, expresses a form of the GCL protein for the purpose of synthesizing GSH

Dysregulation of GCL enzymatic function and activity is known to be involved in the vast majority of human diseases, such as diabetes, Parkinson's disease, Alzheimers disease, COPD, HIV/AIDS, and cancer. This typically involves impaired function leading to decreased GSH biosynthesis, reduced cellular antioxidant capacity, and the induction of oxidative stress.

Measuring GR, GPx, GCL in Autism

Fortunately somebody has already measured GR, GPx and GCL in autism, and not surprisingly they are all dysfunctional.  The paper is by the Chauhans, who already feature on my Dean’s list of researchers.

Impaired synthesis and antioxidant defense of glutathione in the cerebellum of autistic subjects: alterations in the activities and proteine xpression of glutathione-related enzymes.

In the cerebellum tissues from autism (n=10) and age-matched control subjects (n=10), the activities of GSH-related enzymes glutathione peroxidase (GPx), glutathione-S-transferase (GST), glutathione reductase (GR), and glutamate cysteine ligase (GCL) involved in antioxidant defense, detoxification, GSH regeneration, and synthesis, respectively, were analyzed. GCL is a rate-limiting enzyme for GSH synthesis, and the relationship between its activity and the protein expression of its catalytic subunit GCLC and its modulatory subunit GCLM was also compared between the autistic and the control groups. Results showed that the activities of GPx and GST were significantly decreased in autism compared to that of the control group (P<0.05). Although there was no significant difference in GR activity between autism and control groups, 40% of autistic subjects showed lower GR activity than 95% confidence interval (CI) of the control group. GCL activity was also significantly reduced by 38.7% in the autistic group compared to the control group (P=0.023), and 8 of 10 autistic subjects had values below 95% CI of the control group. The ratio of protein levels of GCLC to GCLM in the autism group was significantly higher than that of the control group (P=0.022), and GCLM protein levels were reduced by 37.3% in the autistic group compared to the control group. A positive strong correlation was observed between GCL activity and protein levels of GCLM (r=0.887) and GCLC (r=0.799) subunits in control subjects but not in autistic subjects, suggesting that regulation of GCL activity is affected in autism. These results suggest that enzymes involved in GSH homeostasis have impaired activities in the cerebellum in autism, and lower GCL activity in autism may be related to decreased protein expression of GCLM.

GCLM referred to above is Glutamate-cysteine ligase, it is the first rate limiting enzyme of glutathione synthesis, it is encoded by the GCLM gene. This is an enzyme/ gene you would want to upregulate.

https://en.wikipedia.org/wiki/GCLM

Fortunately we can upregulate GPx enzyme activity with catechin or epicatechin.

Effects of catechin and epicatechin on superoxide dismutase and glutathione peroxidase activity,  in vivo

  

Abstract

OBJECTIVES:

The objective of this study was to investigate the effects of catechin and epicatechin on the activity of the endogenous antioxidant enzymes superoxide dismutase (SOD) and glutathione peroxidase (GPx) (as well as the total antioxidant capacity (TAC)) of rats after intra-peritoneal (i.p.) administration.

METHODS:

Twenty-four Wistar rats were randomly divided into two groups: the experimental group which was administered daily with a 1:1 mixture of epicatechin and catechin at a concentration of 23 mg/kg body weight for 10 days and the control group which was injected daily with an equal amount of saline. Blood and urine samples were collected before and after the administration period, as well as 10 days after (follow-up).

RESULTS:

Intra-peritoneal administration of catechins led to a potent decrease in GPx levels and a significant increase in SOD levels. TAC was significantly increased in plasma and urine. Malonaldehyde levels in urine remained stable. In the animals treated with catechins, SOD activity showed a moderate negative correlation with GPx activity.

DISCUSSION:

Boosting the activity of the antioxidant enzymes could be a potential adjuvant approach for the treatment of the oxidative stress-related diseases.

The objective of this study was to determine whether i.p. administration of catechin and epicatechin could affect the activity of the antioxidant enzymes, SOD and GPx, as well as the TAC in RBCs, blood plasma, and urine.

The antioxidant enzymes are agents that promote reactions for the removal of reactive species (e.g. O2•,.H2O2, etc.). They constitute the first line of

defense against oxidative stress. In conditions of increased oxidative stress, the upregulation of the enzyme activity or even, a possible protection of the enzymes’ substrate could be of great importance.

Oxidative stress disturbing homeostasis can be resolved by the application of catechins and epigallocatechin gallate (EGCG)18 and there is growing evidence that, the protection, offered by flavonoids and their in vivo metabolites, is not mediated primarily by H-donating antioxidant processes, but is likely to be partly mediated through specific actions, within signaling pathways.

Catechin and epicatechin administration modulated the activity of SOD and GPx but the overall TAC of the RBCs and of the rat’s plasma remained stable.

Catechins are considered as potent antioxidants and many of their biological actions have been attributed to that. It would have been expected that since catechins are potent antioxidants in vitro, they would have exerted their classical hydrogen-donating antioxidant activity leading to an increase in TAC; as it is seen in the TAC of plasma. The modulation of the enzymes activity may provide evidence that, catechins exert their primary antioxidant activity by specific action within specific molecular pathways, rather than as scavengers of free radicals.

Oxidative stress is a prominent feature of many acute and chronic diseases and even of the normal aging process. The normal function of the antioxidant enzymes guarantees the preservation of cell integrity and thus they can be considered as potential therapeutic targets of oxidative stress-related diseases.

Various antioxidants are available for therapeutic use but most of them have failed in clinical studies of diseases correlated with oxidative stress. Our results suggest that catechins exert their activity not only by H-donating antioxidant processes but likely through mechanisms and pathways that directly or indirectly regulate the expression of the enzymatic antioxidants.

The understanding of these pathways could be important, in developing pharmacological strategies against oxidative stress-related diseases.

For those with autism plus GI issues / ulcerative colitis :- 

  

  

Epicatechin Used in the Treatment of Intestinal Inflammatory Disease: An Analysis by Experimental Models

Abstract

Background. This study was pathway of (−)-epicatechin (EC) in the prevention and treatment of intestine inflammation in acute and chronic rat models. Methods. Intestine inflammation was induced in rats using TNBS. The morphological, inflammatory, immunohistochemical, and immunoblotting characteristics of colon samples were examined. The effects of EC were evaluated in an acute model at doses of 5, 10, 25, and 50 mg/kg by gavage for 5 days. The chronic colitis model was induced 1st day, and treated for 21 days. For the colitis relapse model, the induction was repeated on 14th. Results. EC10 and EC50 effectively reduced the lesion size, as assessed macroscopically; and confirmed by microscopy for EC10. The glutathione levels were higher in EC10 group but decreased COX-2 expression and increased cell proliferation (PC) were observed, indicating an anti-inflammatory activity and a proliferation-stimulating effect. In the chronic colitis model, EC10 showed lower macroscopic and microscopic lesion scores and increase in glutathione levels. As in the acute model, a decrease in COX-2 expression and an increase in PC in EC10, the chronic model this increase maybe by the pathway EGF expression. Conclusion. These results confirm the activity of EC as an antioxidant that reduces of the lesion and that has the potential to stimulate tissue healing, indicating useful for preventing and treating intestine inflammation.

Inhibition of ulcerative colitis in mice after oral administration of a polyphenol-enriched cocoaextract is mediated by the inhibition of STAT1 and STAT3 phosphorylation incolon cells.

Abstract

We studied a polyphenol-enriched cocoa extract (PCE) with epicatechin, procyanidin B2, catechin, and procyanidin B1 as the major phenolics for its anti-inflammatory properties against dextran sulfate sodium (DSS)-induced ulcerative colitis (UC) in mice. PCE reduced colon damage, with significant reductions in both the extent and the severity of the inflammation as well as in crypt damage and leukocyte infiltration in the mucosa. Analysis ex vivo showed clear decreases in the production of nitric oxide, cyclooxygenase-2, pSTAT-3, and pSTAT1α, with NF-κB p65 production being slightly reduced. Moreover, NF-κB activation was reduced in RAW 264.7 cells in vitro. In conclusion, the inhibitory effect of PCE on acute UC induced by DSS in mice was attenuated by oral administration of PCE obtained from cocoa. This effect is principally due to the inhibition of transcription factors STAT1 and STAT3 in intestinal cells, with NF-κB inhibition also being implicated.

 Here is an excellent paper on oxidative stress.  It is about COPD, but applicable to any condition in which oxidative stress is present.

Oxidative stress and free radicals in COPD – implications and relevance for treatment

  

The following paper would suggest that people with COPD would benefit from epicatechin.

The cocoa flavanol (-)-epicatechin protects the cortisol response.

Abstract

Various health benefits of the cocoa flavanol (-)-epicatechin (EC) have been attributed to its antioxidant and anti-inflammatory potency. In the present study we investigated whether EC is able to prevent deterioration of the anti-inflammatory effect of the glucocorticoid (GC) cortisol in the presence of oxidative stress. It was found that cortisol reduces inflammation in differentiated monocytes. Oxidative stress extinguishes the anti-inflammatory effect of cortisol, leading to cortisol resistance. EC reduces intracellular oxidative stress as well as the development of cortisol resistance. This further deciphers the enigmatic mechanism of EC by which it exerts its anti-inflammatory and antioxidant action. The observed effect of the cocoa flavanol EC will especially be of relevance in pathophysiological conditions with increased oxidative stress and consequential GC resistance and provides a fundament for the rational use of dietary antioxidants.

Conclusion

It would seem that in someone with autism epicatechin is worth a try, other indicators might well include:-

·        Low MOCOS

·        Low ferritin

·        Oxidative stress

And even

·        Restless leg syndrome (symptom of low ferritin)

·        Poor sleep patterns (symptom of low ferritin)

Boosting anti-oxidant enzymes (via gene expression) may be a useful add-on therapy to anti-oxidants themselves.  This is likely true for COPD and autism/schizophrenia.

If you are wondering whether there is anemia or iron deficiency in autism, your questions are likely answered here:-

Iron deficiency in autism and Asperger syndrome.

This research considers the prevalence of iron deficiency in children with autism and Asperger syndrome and examines whether this will influence guidelines and treatment. Retrospective analysis of the full blood count and, as far as available, serum ferritin measurements of 96 children (52 with autism and 44 with Asperger syndrome) was undertaken. Six of the autistic group were shown to have iron deficiency anaemia and, of the 23 autistic children who had serum ferritin measured, 12 were iron deficient. Only two of the Asperger group had iron deficiency anaemia and, of the 22 children who had their serum ferritin measured, only three were iron deficient. Iron deficiency, with or without anaemia, can impair cognition and affect and is associated with developmental slowing in infants and mood changes and poor concentration in children. This study showed a very high prevalence of iron deficiency in children with autism, which could potentially compromise further their communication and behavioural impairments.

As we saw with biotin and soon will with vitamin B12, it seems that people with autism can have unexpected deficiencies of key substances even though their diet may not be deficient.  The identified iron deficiency is an iron storage deficiency.  With biotin the body was unable to recycle the vitamin biotin, due to a problem with the enzyme biotinidase, hence there was a deficiency.

Correcting these deficiencies is quite simple and may well improve any related autism symptoms.  In people without these dysfunctions/deficiencies any such supplements would yield no benefit and might even produce side effects.