= Autism & ADD

SCIENTIFIC PROOF: MILK CAUSES AUTISM & ADD

Florida researcher, Robert Cade, M.D., and his colleagues have identified a milk protein, casomorphin, as the probable cause of attention deficit disorder and autism. They found Beta-casomorphin-7 in high concentrations in the blood and urine of patients with either schizophrenia or autism.

(AUTISM, 1999, 3)

Eighty percent of cow's milk protein is casein. It has been documented that casein breaks down in the stomach to produce a peptide casomorphine, an opiate.

Another researcher observed that casomorphin aggravated the symptoms of autism.

(Panksepp, J. Trends in Neuroscience, 1979, 2)

A third scientist produced evidence of elevated levels of endorphin-like substances in the cerebro-spinal fluid of people with autism.

(Gillberg, C. (1988) Aspects of Autism: Biological Research Gaskell:London, pp. 31-37)

The Autism Research Unit, School of Health Sciences has the following information on their website:

"The quantities of these compounds, as found in the urine, are much too large to be of central nervous system origin. The quantities are such that they can only have been derived from the incomplete breakdown of certain foods."

http://osiris.sunderland.ac.uk/autism/

PARENT'S MAGAZINE REPORTS MILK LINK TO AUTISM

Search the Internet and you'll find many anecdotal stories from parents blaming their children's autism on milk and dairy products. One such story appeared in the February, 2000 issue of "Parent's Magazine."

http://www.livingsensibly.org/autism.html

See references 25 to 33
____________________

A Chemical Aetiology for Autistic Spectrum Disorders:
Opioid Peptides and Secretin
Unpublished Paper by Stephen Dealler

http://trainland.tripod.com/stephen.htm

The suggestion that the syndrome of autism was similar in psychological
terms to that seen in children who had received morphine was put forward
by Panksepp in 1978 (1). The reason as to how this might take place in
vivo when no morphine was being administered and no poppy derived
products were present in their diet was initially unclear. The finding
that casomorphins produced a reduction in distress by chicks when
separated from others (2) was considered as evidence.

Increased levels of a group of urinary peptides were found
(3,4,5,6,7,8,9) and demonstrated by gel filtration, HPLC, and SDS-PAGE
electrophoresis (10) These results were not found in the urine of
children with the fragile X syndrome (4) and a different profile of
peptide sizes was seen in late onset infantile autism, neonatal-onset
infantile autism, and mixed onset cases including atypical ones (4).
False-positive urinary profiles were found in 5-7% of controls and false
negative ones in 5% (n=126) over 12 years. It has been known for some
time that if peptidases are lacking in the gut then there is an increase
in peptides found in the urine (11) but no histopathological changes
have been reported in autism and no absolute lack of peptidases. It is
suggested, however that the mechanism by which the peptides enter the
blood is as a result of the lack of activity of two specific peptidases
(4) as this fits with the genetic rate for the disease in siblings.
There had been some indication of gut abnormality in that an excess of
cases of coeliac disease was found (4) by one group and indications of
malabsorption in others (12,13,14,15). Good research has shown that
there is an abnormal intestinal permeability in children with autism
(16) and hence the uptake of these peptides is not necessarily the
result of the lack of specific peptidases at all.

The hypotheses as to why opioid peptides should act to produce an
autistic syndrome have been fully described in a chemical manner
(6,17,18,19,20) but inadequate information is available to be certain of
the mechanisms described.

It was discovered that these individual peptides may have opioid
activity (21,22,23) and certainly had biological activity of some kind
(24). The finding that some had opioid-like receptor binding (24,25) was
followed up by the finding that bovine casomorphins, opioid peptides
from milk, were present in urine and dialysis fluid (4,25). It is known
that casomophins and gluteomorphins are produced in the gut by
proteinase digestion of milk and wheat proteins (26,27,28) and that
casomorphins can have been involved in post partum psychosis (29) and
hence penetrate the CNS (30).

An increase in IgA antibodies was found in the blood of autistic
children against gluten and casein (4) but in 4 of the 44 children no
such antibodies were found whatever. This made the value of immune
testing difficult to understand: there might have been a large enough
absorption for the IgA to have interacted with the peptides and taken
out of the circulation or only a small uptake; in which case antibodies
were formed but are of little significance.

When put onto a diet without milk or gluten-containing foods it can be
shown that peptide excretion decreases in the urine from around
30mmol/24hrs to around 10mmol/24hrs (4,31) and when the diet was stopped
the peptide excretion increased to previous levels.

When first started on this diet, it was noted that some of the children
showed a change in their psychological activity and were 'cold turkey'
at some points over the first few days, suggesting an opioid withdrawal
effect. Autistic patients put on a diet without gluten or milk protein
improved clinically (4,5,31,32,33) and this was shown using
psychological tests, tests of teaching, and indications by parents.
Although it is difficult to explain the figures used to describe the
improvement without understanding the mechanism, it is clear that an
improvement did take place.

A diet of this kind is difficult to maintain and a number of patients
withdrew from the studies after having shown an improvement. These
patients showed a slow regression to the previous condition (4)

Autistic children (12) were tested with naltrexone, a long acting opiate
inhibitor, in a double blind controlled trial and showed an overall
improvement in their condition. The partial agonist activity of
naltrexone was a problem in that higher doses appeared to have a lesser
effect than lower ones (34). Also, the naltrexone did not alter the
behaviour of the patient simply by decreasing all autistic action but
rather by modifying certain ones. A reduction in stereotypies,
increasing in verbal production, an improvement in social behaviour and
self-injurious behaviour. Other, open trial studies, showed similar
results (35-42). Overall the results were not as great as were hoped and
when naltrexone was stopped, previous behaviour returned.

It has been suggested that the long term exposure of the brain to
peptide morphines from the diet would have a trophic effect on the brain
in the same way as morphine itself (28,29,43) and that this may explain
the progressive nature of autism in some cases; giving rise to an
increase in epileptic fits and EEG abnormalities in increasing age (44).
The long term endorphins and naltrexone were separately shown to modify
the development of the brain (17,45). No research has been carried out
currently to see what effect long term removal of these diet-derived
peptides may have and whether any of the damage may recover. There have
been individual reports of epilepsy ceasing following the removal of
specific factors from the diet but the mechanisms are unclear.

Secretin

Secretin is a 27 amino acid polypeptide discovered in 1902 by Bayliss
and Starling (46). Compared to other neuropeptides it has been poorly
researched, with publications reaching a peak between 1980 and 1985
(47). Its aminoacid sequence was not found until 1965 (48) because of
the low quantities that were present to test. However it was first
synthesised shortly afterwards (49). The structure turned out to be well
retained through evolution; both beef and porcine (50) secretin differed
from human secretin by 2 amino acids and dog secretin differed by a
single one (51). When formed in the rat it is made as a precursor
protein intracellularly from a 739 base gene, of which only 692 bases
are expressed as mRNA (52). The precursor protein exists as a signal
peptide, an amino terminal peptide, secretin, and a carboxy terminal
peptide. The signal, amino and carboxy terminal peptides are cleaved
from secretin before its release. This precursor and gene are identical
in the duodenum and in the brain.

Secretin is one of a group of neuropeptides that were originally felt to
be found only in the gut and to act as endocrines. All of them have
been found to be present in the brain also. They are known as the
secretin group of peptides (table 1). This is important as research into
all of these has been greater than into secretin itself (7) and certain
factors from this research are expected to be significant for secretin.

This group of neuropeptides is remarkable in that, although they are
similar in structure, they have often quite different activities and
different receptors in tissues. Some of them will cross react between
receptors but the cross reaction is minimal in vivo despite this
similarity (52a). It is likely that evolutionary change has demanded
specific action even as a result of small changes in the peptide
structure. As a result it would be expected that only small changes in
secretin structure would decrease its activity but possibly not alter
its assay using standard radio-immunoassay techniques. Because of this
it is essential that if any assay of secretin is undertaken, it is
necessary to test the activity of the hormone and its structure: not
just the presence of the peptide.

Secretin mode of action

Secretin interacts with highly specific cellular surface receptors which
carry intracellularly a Gs type protein which then changes the action
of
certain cellular enzymes. Human secretin receptors belong to a seven-
transmembrane domain subfamily that includes receptors for VIP, PACAP
and glucagon. The receptor structures have been well worked out (53) as
being generally around 1616 bases (54) in length (53) and distinct
receptors were characterized in the guinea pig pancreatic acini (54),
rat gastric glands (55), rat cholangiocytes (56), mouse neuroblastoma
cells (57), and mouse-rat NG108-15 neuroblastoma-glioma hybrid cells
(58). The rat secretin receptor (RSR) cDNA was the first to be worked
out as a member of a distinct new family of G protein-coupled receptors
(59) that now include VIP (60,61,62), PACAP (63,64,65), glucagon
(66,67), parathyroid hormone (68) and calcitonin (69). It can be shown
that rat and human receptors are similar; being closely homologous
except at the amino and carboxy terminals (53). The main action seen is
the increase in adenyl cyclase activity and it is thought that this is
the major mode of action of secretin both in the periphery and in the
central nervous system. There is also an increase in calcium inside the
affected cell and an increase in inositol phosphate production. A
modified form of secretin has been shown to act as an antagonist in vivo
in rodents due to its ability to interact with the receptor but in some
way not stimulate its activity (49)

Secretin activity in the periphery

Secretin is only produced peripherally by the S cells of the proximal
section of the duodenum and acts as a feed back loop used to counter the
stomach acid arriving there after food. The presence of acidic fluid in
the duodenum or the distension of it causes secretin release. This is
passed though the circulation to acinar cells of the pancreas where it
causes an increase in the release of alkali. It causes an increase in
fluid release from the biliary system, the small intestine epithelium
and the pancreas. It decreases the production of gastric acid, gastrin
(69), but increases the release of pepsinogen from gastric chief cells.
Some cardiac and renal activity is seen but this has not been fully
investigated. Secretin also causes an increase in the activity of
tyrosine hydroxylase (TH) in cervical ganglion neurones and
phaeochromocytoma cells (70,71). This enzyme is the limiting section of
the manufacture of catecholamines (e.g. adrenaline) and hence it is felt
that some adrenergic activity may be seen as a result but it has not
been quantified.

TH activity has been shown to be subject to the regulation by the cAMP
system inside the cell as well as calcium and cGMP second messenger
systems (70). Treatment of intact rat PC12 cells with neuropeptides
including secretin stimulated TH activity 2 to 3 fold (71).

This activity is not surprising in that receptors for secretin have been

found to be present in the pancreas, kidney, rat gastric glands,
cholangiocytes, neuroblastoma cells, neuroblastoma glioma cells, lung,
and intestinal epithelium. All of these appear to have more receptor
activity than that seen in brain, heart or ovary. However no receptors
at all are seen in other tissues tested. Cross reactivity between the
secretin group peptides has been seen but to a low degree. Measured KD
levels were 10-7 to 10-10 mol.

In neonatal animal experimental secretin has been shown to be involved
in the growth and development of the gut, stomach and pancreas (72,73).

Secretin activity in the brain

The quantity of secretin in neurological tissues seems to be similar to
that in the duodenum, varying between 7pg/mg (wet weight) of the cortex
to 130 pg/mg of the pineal (74) (table 2).

It was first demonstrated in the brain of the rat and pig (but not in
the guinea pig)(75). Highest levels were found in the medulla oblongada,
thalamus, hypothalamus olfactory bulb, hippocampus, midbrain, cerebellum
and brain stem (74,80) Initially there was difficulty in demonstrating
that this was correct and researchers failed

to repeat the findings. However the demonstration of mRNA for the
precursor peptide in similar quantities to that found in the duodenum in
the same tissue ratios as found for the secretin itself means that this
can no longer be open to argument.

Like other transmitters such as dopamine, the action of peptides may be
through the stimulation of adenylate cyclase thus increasing levels of
cyclic AMP to cause intracellular changes rather than alterations in
transmembrane potentials (70,76,77) as is seen with acetyl choline.

Specific activity has been shown by secretin: inhibition of prolactin
and leutinising hormone release; displacement of VIP from certain
receptors; increase the turnover and level of dopamine; increase in
adenyl cyclase activity in the hypothalamus; glycogenolysis in primary
culture glial cells; enhancement of pancreatic volume and bicarbonate
response to acid in the duodenum (this was shown not to be a systemic
effect of secretin escaping from the intracerebral inoculation of the
drug into the blood) (78)

Penetration of the blood brain barrier

One of the major questions in this field has been whether or not
secretin and other neuropeptides could be used as therapeutic agents.
Around 50% that have been tested have been found to cross the blood
brain barrier easily and rapidly with levels appearing in the CSF within
5 minutes of peripheral inoculation (79). Secretin has not been tested
in this respect but, as all of the other members of the group seem to
penetrate the BBB, it is unlikely that it is not true for secretin (80).

The finding that i.v. secretin caused a rapid increase in prolactin
whereas i.c. it caused a decrease, has suggested that there is a short
negative feed back loop present and that this probably is taking place
in the hypothalamus. This also suggests that the BBB is penetrated by
secretin and reaches some specifically sensitive site. Further work must
be carried out into this, however.

It should also be remembered that intraventricularly administered
peptides also appear rapidly in the blood (81) although, again this
experiment was not carried out with secretin.

Lack of secretin activity as a potential cause of the gut uptake and
inadequate breakdown of opioid peptides in autistics

The genetic aspects of autism suggest that either two gene abnormalities
are required or, as there is a 36%-91% correalation in monozygotic twins
(82,83,84) it would suggest an approximately 50% penetration of a single
gene. The finding of a dizygotic sibling concordance rate of 3% is
similar to that seen in a condition that required two recessive
alterations (6.3%), rather than the 50% penetration of a single one,
which would appear in 12.5%.

The finding that the gut in autistics may be abnormal in that it takes
up long chain molecules (16), may suggest that it is not so much the
peptidase acitivity that is inadequate but that there is a physiological
barrier between the gut and the blood that is ineffective. This has
been shown with carbohydrates and with peptides.

It has been shown that secretin is relatively high in the blood of
neonates (72,73) and that infusions of it causes precocious cessation of
intestinal macromolecular transmission (72).

The clinical effect seen from secretin in autism is strange in that the
action has been reported as descriptions from patients' relatives to
increase for several days after a single intravenous injection and the
effect is seen to reach a peak between 2 and 4 weeks. This cannot be
simply due to the effect of secretin on the brain as a neurotransmitter
as the half-life of compound is short. This long term effects suggested
would fit better with secretin as an inducer of protein production or
cellular maturation. causing an alteration in cells that would either be
themselves destroyed or lose the action gradually. Although secretin
when inoculated intrecerebrally was shown to induce morphine action,
this effect did not last for long periods and hence cannot be thought of
as the specific cause of the phenomenon in autism (85). For instance,
one hypothesis could be that the action of the injected secretin may be
as an inducer of changes in the gut epithelium cells that are themselves
lost by shedding over the following weeks.

If secretin was seen to prevent the pathological gut physiology seen in
some autistic children then this effect might be seen if inadequate
secretin was being produced by the S cells of the duodenum or if the
secretin that was produced did not interact adequately with the receptor
molecules (this might be found if only small changes were made). Some
autistic cases may not respond at all to secretin and either the
mechanism by which it would act was faulty (e.g. the secretin receptors)
or some other pathology was present to which secretin was not
significant. Also, it must be remembered that alterations in the
receptor may give rise to inadequate action taking place on the G-
protein and from there a lack of change in cellular adenyl cyclase
activity.

If this chain of action gave rise to the lack of secretin physiology in
the autistic child, then it would not be found that all would respond to

injected hormone. Only those in which the secretin was missing or
altered would improve. Any cases in which it was either the receptor or
its mode of action that were ineffective would not respond.

Psychology

It may also be worth discussing how, in psychological terms, disruptions
of opioid mechanisms might lead to the symptoms of autism. There is
convincing evidence that autistic spectrum disorders are associated with
what are termed theory-of-mind problems (86, 87, 89). This means that
autistic children appear unable to comprehend the mental states of other
people or appreciate their perspectives. This specific pattern of
psychological problems is associated with other, still quite specific,
problems with what is termed central executive functioning (90,88). A
convincing argument can be developed to the conclusion that specific
deficits in central executive functioning, probably the ability
simultaneously to process more than two pieces of information, lead to
the problems observed in autistic spectrum disorders. It is entirely
possible that secretin and other opioid peptides mediate these
processes.

Discussion

The reports that secretin appears to improve a group of autistic
children and that this takes place over a period much greater than it
would be expected due to its short term action as a neurotransmitter and
to induce the release of alkali into the duodenum. The long term
hormonal action of secretin has been poorly investigated but may involve
gut development. The finding that secretin inoculated into 3 autistic
children (91) caused an excessively large increase in alkali secretion
would be consistent with there being inadequate secretin present
normally. While the long term hormonal actions of secretin are
inadequately studied, further research needs to be carried out
concerning autism; first to see if secretin has adequate action in
autism at all (92) but if it does, however small, to follow its mode of
action as a key to the physical and chemical pathology of the condition
as yet poorly understood.

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For children with autism, milk may very well be the major factor. One out of five American children have been diagnosed with attention deficit disorder. One out of five American children take Ritalin. An alternative therapy? NOTMILK!