Re: New study that validates vitamin E!!!!!

February 23, 2008

In a message dated 2/23/2008 5:43:38 P.M. Eastern Standard Time,

lizlaw@... writes:

Not sure why some of you area jumping on the fact that it may

> not 'just' be mercury that's an issue. I'm very aware that I pulled

> it out of context which was in direct response to the (?) message

> from Liz that mercury causes autism so what causes apraxia. Anyone

> who doesn't know that apraxic children were also looked at but not

> mentioned is not in reality. Carolyn below in the archives for

> example was one of Dr. Ming's patients and I know that they did

check

> at that time cheek cell scrapings from all -but in the end the

> funding reasons typically 'only' mentions autism which is very sad

for the

> even larger number of apraxic children. Also... I left the entire

> published paper intact right underneath where one can learn that

> there are other toxins to worry about. In this group the toxin

issue

> has been up for conversation for years

> _http://www.cherab.http://wwhttp://ww_

(http://www.cherab.org/news/Save.html) -and Dr. Xue Ming from this

> study was one of the MDs I was asked to present to at UMDNJ back in

> 2002 on my theory of how toxins cross the placenta while we are

> pregnant. Here's just one post about it from then:

I'm not sure whether my thoughts mean much or anything-- but I've never been

exposed to mercury during pregnancies-- we don't vaccinate, and I personally

was never exposed to any mercury, so for MY son's pregnancy-- this theory of

mercury causing Apraxia-- wouldn't be valid.

Becky

**************Ideas to please picky eaters. Watch video on AOL Living.

(http://living.aol.com/video/how-to-please-your-picky-eater/rachel-campos-duffy/

2050827?NCID=aolcmp00300000002598)

February 23, 2008

So E is effective against mercury induced oxidative stress in

autistics. Would the theory for apraxics be that E is effective

against oxidative stress of as yet unknown origin?

Glad to see this. This campus is down the street from my house. I may

just be able to get more help. Thanks!

>

> American Journal of Biochemistry and Biotechnology 4 (2): 218-225,

> 2008

> ISSN 1553-3468

> Corresponding Author: C. Wagner, Psychology, Busch Campus,

> Rutgers University, New Brunswick, NJ 08854

> Tel: 732-445-4660 Fax: 732-445-2263

> 218

> Evidence of Oxidative Stress in Autism Derived from Animal Models

> 1Xue Ming, 2 A. Cheh and 2 L. Yochum,

> 3Alycia K. Halladay and 2 C. Wagner

> 1Pediatric Neuroscience, UMDNJ, Newark, NJ

> 2Psychology, Rutgers University, New Brunswick, NJ

> 3Autism Speaks, Princeton, NJ

> Abstract: Autism is a pervasive neurodevelopmental disorder that

> leads to deficits in social

> interaction, communication and restricted, repetitive motor

> movements. Autism is a highly heritable

> disorder, however, there is mounting evidence to suggest that

> toxicant-induced oxidative stress may

> play a role. The focus of this article will be to review our animal

> model of autism and discuss our

> evidence that oxidative stress may be a common underlying mechanism

> of neurodevelopmental

> damage. We have shown that mice exposed to either methylmercury

> (MeHg) or valproic acid (VPA) in

> early postnatal life display aberrant social, cognitive and motor

> behavior. Interestingly, early exposure

> to both compounds has been clinically implicated in the development

> of autism. We recently found

> that Trolox, a water-soluble vitamin E derivative, is capable of

> attenuating a number of

> neurobehavioral alterations observed in mice postnatally exposed to

> MeHg. In addition, a number of

> other investigators have shown that oxidative stress plays a role

in

> neural injury following MeHg

> exposure both in vitro and in vivo. New data presented here will

show

> that VPA-induced

> neurobehavioral deficits are attenuated by vitamin E as well and

that

> the level of glial fibrillary acidic

> protein (GFAP), a marker of astrocytic neural injury, is altered

> following VPA exposure. Collectively,

> these data indicate that vitamin E and its derivative are capable

of

> protecting against neurobehavioral

> deficits induced by both MeHg and VPA. This antioxidant protection

> suggests that oxidative stress

> may be a common mechanism of injury leading to aberrant behavior in

> both our animal model as well

> as in the human disease state.

> Key words: Vitamin E, trolox, valproic acid

> INTRODUCTION

> The core symptoms of autism include language

> deficits, impaired social interactions and inappropriate,

> stereotypic and sometimes self-injurious behaviors. The

> etiology of autism remains unknown but may involve

> early exposure to environmental toxicants acting upon

> genetically-sensitive individuals. No single toxicant has

> been identified; rather a broad range of toxicants

> including drugs, metals, solvents, herbicides, pesticides,

> etc. have been associated with autism[1-3]. A common

> feature across this range of potential compounds is

> toxicant-induced oxidative stress causing neuronal

> damage leading to the behavioral phenotype of

> autism[4-6]. Likewise, no single gene has been identified

> but, rather, a constellation of as many as 15

> polymorphisms may ultimately predispose the

> individual to autism. Again, genetic alterations leading

> to compromised handling of toxicant-induced reactive

> oxygen species (ROS) has been a common theme.

> Since the etiology of autism is unknown, it is

> essential that animal models be developed. The

> behavioral symptoms of autism have proven difficult to

> model in other species. Accordingly, we have initiated

> work on a novel strategy to model the behavioral

> phenotype of autism in mice[1]. In this model, the

> normal development of key behaviors is carefully

> monitored from birth through adolescence. Once the

> maturation of these key behaviors is understood in

> terms of the postnatal day(s) of life in which subjects

> are able to successfully perform the task or engage in

> the behavior, the performance of mice with early

> toxicant exposure and/or genetic modification can be

> assessed.

> The model strategy begins by characterizing

> behavioral manifestations of developmental disorders

> as retardations (a behavior fails to develop during a

> critical period of maturation), regressions (a behavior

> develops at about the right time but then is lost with

> Am. J. Biochem. & Biotech., 4 (2): 218-225, 2008

> 219

> later development, especially following toxicant

> exposure), or intrusions (the appearance of behaviors

> aberrant in form or frequency which mask normal

> development). Most developmental disorders include

> some combination of these conditions. In this

> framework, the hypothesis that environmental toxicants

> or genetic alterations are causally involved in autism

> can be readily tested. That is, acute or repeated

> exposure to a toxicant should disrupt neurobehavioral

> development causing behavioral retardation, regression,

> or intrusions and these toxicant-induced behavioral

> deficits should occur at lower doses in the geneticallysensitive

> mice. Traditionally, animal models of

> developmental disorders have not examined these three

> scenarios of retardations, regressions and/or intrusions

> but, instead, focus on single aspects of neurobehavioral

> development. The judicious use of toxicants associated

> with autism or toxicants known to damage brain regions

> associated with autism confers some selectivity of the

> model for autism. Likewise, manipulation of genes

> associated with autism also confers some selectivity of

> this model for autism. Finally, administering a battery

> of tests that assess social, cognitive and motor

> maturation of the mice confers some selectivity for

> autism. Ultimately, it is the possibility of combining

> select toxicant exposure in genetically-sensitive mice

> followed by thorough assessment of social, cognitive

> and motor skill maturation that makes this a

> comprehensive animal model of autism.

> In our initial studies, we identified toxicant induced

> retardation of motor and cognitive skills following pre-

> or post-natal exposure to sodium valproate (VPA).

> Likewise, we were able to demonstrate dramatic loss of

> acquired skills, i.e. regressions, following post-natal

> VPA administration[1]. Finally, we demonstrated

> toxicant induced intrusions wherein toxicant-treated

> subjects exhibited dramatic increases in stereotypic and

> self-injurious behaviors akin to those seen in autism[7,8].

> VPA was chosen as our first agent to test this

> model following reports of an association between

> autism and prenatal exposure to this teratogen[9-13].

> Previous studies have also demonstrated impairment in

> cognitive, motor, attention and social development in

> rats administered pre- or post-natal VPA[14-17].

> Accordingly, in our first studies[1] mice were exposed to

> VPA either in utero or post-natally. The prenatal

> exposure time reflected a period of cerebellar Purkinje

> cell generation differentiation in the mouse[10,14,17,18].

> The post-natal time of P14 was based on our

> observation that critical cerebellar-mediated behaviors

> of mid-air righting and negative geotaxis mature or first

> appear on this day in the mouse[1] and because of

> continued neuronal and glial development in other brain

> regions[15,19,20]. Of importance, VPA administration

> results in high levels of markers for oxidative stress and

> lipid peroxidation including 15-F-isoprostane and

> thiobarbituric acid reactive substances[21-23].

> An organic mercury, MeHg, was selected as our

> second compound for testing because it is an important,

> widely distributed environmental toxicant. MeHg does

> cross the placental barrier and, in humans exposed in

> utero to acute high doses, was shown to cause

> retardation in cognitive and locomotor development

> along with numerous other neurological symptoms

> including seizures and cerebral palsy[24]. Nonetheless, it

> is important to note that autism was not found to be

> associated with either pre- or neonatal exposure to

> organic mercury.

> The consequences of low dose, chronic exposure to

> mercury through fish consumption are somewhat more

> controversial with some studies showing deleterious

> effects while others show no adverse consequences[2,24].

> Early exposure to mercury has been shown to disrupt

> the neurobehavioral development of other species

> including rodents and primates[25]. The mechanism

> through which MeHg exerts its toxicity is thought to be,

> in part, mediated by disruption of neural cell adhesion

> molecules[26]. In addition, oxidative stress is involved in

> MeHg-induced neurotoxicity as demonstrated by

> increased ROS and thiobarbituric acid reactive

> substances and a reduction in GSH levels[27]. In

> addition, the neurotoxicity of MeHg in cultured neurons

> was blocked by the pretreatment with antioxidants[28].

> Trolox, a water-soluble derivative of vitamin E,

> protects against MeHg-induced neurotoxicity in rats[29].

> Likewise, antioxidants produced protective effects

> against MeHg toxicity in cultured human neurons and

> astrocytes[30]. Indeed, ROS have been implicated in

> MeHg-induced neurotoxicity in multiple experimental

> models[27,31-34]. Finally, we have recently demonstrated

> that pretreatment with Trolox protects mice against the

> neurobehavioral deficits induced by postnatal MeHg[8].

> Collectively, these data indicate that early exposure to

> MeHg causes neurobehavioral deficits consequent, at

> least in part, to the generation of ROS.

> In summary, wide ranges of toxicants and genetic

> alterations have been associated with autism. The

> toxicants are thought to have a common mechanism of

> generating ROS[4-6] while the genetic alterations are

> thought to result in enhanced sensitivity to the

> deleterious effects of ROS. Accordingly, we now

> hypothesize that autism may be the result multiple

> exposures to any of a number of toxicants; the initial

> exposure sensitizes the subject such that later exposures

> to the same or different toxicants results in an enhanced

> Am. J. Biochem. & Biotech., 4 (2): 218-225, 2008

> 220

> oxidative stress response. Furthermore, we predict that

> this sensitization will be exacerbated in individuals with

> genetic alterations affecting their handling of ROS. In

> previous studies we have demonstrated a sensitization

> response to dopaminergic toxicants in adult mice

> following prenatal administration of MeHg[7]. We have

> also demonstrated that antioxidant pretreatment protects

> mice against the behavioral deficits induced by early

> exposure to MeHg[8]. Accordingly, the objective of this

> study was to determine if antioxidants administered as a

> pretreatment to VPA would protect the mice against the

> VPA-induced behavioral regression. In addition, we

> sought to determine if the early VPA administration

> would alter levels of glial fibrillary acidic protein

> (GFAP), a marker of astrocytic neural injury, thus

> serving as a biological marker for the VPA-induced

> behavioral deficits.

> MATERIALS AND METHODS

> Subjects: Male and female BALB/c mice (Taconic,

> Germantown, NY) were housed together in plastic

> cages with standard wood chip bedding and free access

> to food and water. All mice were maintained in an

> AAALAC-accredited facility under guidelines set forth

> by the National Institutes of Health. Lights were set on

> a 12 h on: 12 h off cycle and temperature was

> maintained at 25 & #61616;C. Females were checked before 10

> AM for presence of a vaginal plug which was recorded

> as day 0 of embryonic development. Day of birth was

> recorded as day 0 and all pups were labeled for

> individual identification. Body weight was measured

> daily. Female pups were removed from the cage on day

> 5. For the behavioral studies, the sodium valproate

> (Sigma) dose was 400 mg kg & #61485;1 with a saline vehicle and

> the vitamin E dose was also 400 mg kg & #61485;1 but with a

> corn oil vehicle. All injections were s.c. in a volume of

> 1.0 mL 100 & #61485;1 g body weight.

> Negative geotaxis: Negative geotropism was tested on

> postnatal days P13-19 by placing the mouse facing

> downward along a 45 & #61616;C incline. Latency to turn 180 & #61616;C

> such that the head was facing upward along the incline

> was recorded with a maximum of 30 seconds for each

> trial.

> Motor Activity: Motor activity was assessed on days

> P14-19. The chamber consisted of a black

> 42 & #61605;22 & #61605;14 cm Plexiglass box. Six infrared sensors

> placed approximately 7 cm apart and 2.5 cm above the

> floor were used to measure activity over a 10 min

> period.

> Mid-air righting: When a mouse is dropped upside

> down from a height of 45 cm onto a padded surface it

> engages in a mid-air righting reflex with orderly, rostrocaudal

> movements, initiated with head and concluded

> with the hindlimbs such that the animal lands on its

> paws. The behavior first appears on P13 and is fully

> achieved by P17[35]. Mid-air righting has been linked to

> cerebellum development[36]. For the mid-air righting

> test, mice were elevated 45 cm above a foam pad,

> dorsal side down. The animal was released and ability

> to right in mid air assessed scored as the mouse landing

> on its paws on two out of three trials each day. Mice

> were tested on P13-20.

> Protein determination: In order to determine changes

> in protein expression following VPA treatment at

> behaviorally significant time points, animals were

> treated with VPA 600 mg kg & #61485;1 or saline on E13[1] and

> assayed on days P4 and P5 with the cerebellum

> removed and stored at -70 & #61616;C. Protein analysis via gel

> electrophoresis, western blot and densiometry was

> performed according to the methods of Dey et al.[26]

> with some small variations. In summary, whole

> homogenate fractions were homogenized in 1:10 w/v of

> a Tris extraction buffer [50 mM TrisHCl, pH 7.4,

> 0.32M sucrose, 1 mM EDTA, 1 vial to 100 ml protease

> inhibitor (Sigma, St. Louis, MO). The supernatant was

> removed following centrifugation for 10 min at 1000xG

> and combined with equilibration buffer [0.125M

> TrisHCl, pH 6.8, 4% SDS, 20% glycerol, 10%

> mercapoethanol) and immediately heated for 30 min at

> 70 & #61616;C. Protein values were determined using the BCA

> protein assay (Pierce, Rockford, IL) modified for a

> BOBAS FARA II enzyme analyzer (Roche

> Diagnostics, Nutley, NJ). Samples of 10 & #61549;g protein

> were separated by SDS-PAGE on a 10%

> polyacrylamide gradient gel using a Bio-Rad Mini-

> Protean II System (Bio-Rad, Mellville, NY) for GFAP

> and synaptophysin. Proteins were transferred to

> nitrocellulose membranes and were washed twice for

> 10 min each in phosphate buffered saline (PBS) and

> blocked with 5% non-fat dry milk in PBS for 1 h prior

> to application of primary antibody. Immunoblotting for

> GFAP and synaptophysin was performed overnight at

> 4 & #61616;C. All primary antibodies were obtained from Fisher

> (Springfield, NJ). Anitgens were visualized following

> 1 h incubation with secondary peroxidase antibodies

> (Southern Biotechnology Associates, Birmingham, AL)

> and application of chemiluminescence ECL substrate

> detection on Hyperfilm ECL autoradiographic film

> (Amersham). For GFAP detection, this method was

> verified in a separate study using a dose response of

> trimethyltin treatment using a GFAP protein standard

> Am. J. Biochem. & Biotech., 4 (2): 218-225, 2008

> 221

> (Chemicon, Inc.). ECL images were scanned into an

> IBM PC using a Hewlett Packard Scanner with a

> transparency adapter. Densiometric analysis was

> performed using Image Pro Analysis Software using

> percent of saline treated controls as the standard.

> Statistical analysis: All behavioral analysis were

> performed using a repeated measures ANOVA

> including both group, day and sex as main factors, with

> the exception of the mid-air righting response which

> was analyzed using Chi-Square and Fisher's Exact Test.

> RESULTS AND DISCUSSION

> Negative geotaxis: Control mice and those treated with

> vitamin E alone were able to perform the reflexive

> negative geotaxis response, reorienting their head to

> point upward when placed on an inclined plane with

> their head facing down. This reflex improved across

> development, as the latency to re-orient improved

> across testing (F (6, 312) = 5.4, p<0.001). VPA-treated

> mice displayed an increased latency to perform this reorientation

> response (F (1, 52) = 10.0, p<0.005). Posthoc

> analysis revealed that following day 14 treatment

> with VPA, there was a significant regression in the

> performance of this response, which reached statistical

> significance on days 16 and 17. Importantly, this VPAinduced

> regression was blocked by vitamin E

> pretreatment, such that pretreated mice were able to

> perform this response with a similar latency as controls

> on P16 and P17 (F (1, 52) = 5.3, p<0.05). Finally, VPAtreated

> mice regained their ability to perform this

> behavioral response similar to controls by the

> completion of testing on P19 (Fig. 1).

> Negative geotaxis

> Day of testing

> 12 13 14 15 16 17 18 19 20

> Latency to re-orient 180 & #61616;C

> 0

> 5

> 10

> 15

> 20

> 25

> 30 Corn oil/saline

> Corn oil/VPA 400

> Vit E/saline

> Vit E/VPA 400

> *

> Fig. 1: Negative geotaxis: Latency to reorient from

> head down to head up on a 45 & #61616;C incline for

> groups of pups treated with VPA (400 mg kg & #61485;1)

> or saline on P14. Some groups received vitamin

> E pretreatment while others received corn oil.

> *: p<0.05 compared to corn oil/saline

> Mid-air righting

> Day of testing

> 12 13 14 15 16 17 18 19 20

> Percent mice able to right in mid-air

> 0

> 20

> 40

> 60

> 80

> 100

> 120 Corn oil/saline

> Corn oil/VPA 400

> Vit E/saline

> Vit E/VPA 400

> *

> Fig. 2: Mid-air righting: Number of pups successfully

> engaging in mid-air righting (expressed as a

> percent of pups mid-air righting on 2 out of 3

> trials/day) for groups of pups treated with VPA

> (400 mg kg & #61485;1) or saline on P14. Some groups

> received vitamin E pretreatment while others

> received corn oil. *: p<0.05 compared to corn

> oil/saline

> Mid-air righting: Before any treatment was

> administered, less than 20% of the pups were able to

> engage in mid-air writing on P13 but this improved to

> about 75% by P14. This observation is interpreted to

> indicate that cerebellar and general muscular maturation

> have matured by P14. & #61539;2 analysis revealed that

> following VPA-treatment given after behavioral testing

> on P14 caused a regression in mid-air righting on P15

> [ & #61539;2 (3) = 39.8, p<0.0001] when compared to saline

> controls. This regression was still observed on P16 in

> the VPA-treated animals. However, the VPA-induced

> regression was eliminated by pretreatment with vitamin

> E (Fig. 2).

> Motor activity: Mice engaged in a similar amount of

> locomotor activity at the start of testing. This activity

> significantly increased across post-natal development

> [F (1, 38) = 80.2, p<0.001]. Interestingly, there was a

> trend for mice treated with VPA to engage in intrusive

> behaviors following P14 treatment. This was evidenced

> by increased levels of activity across testing, beginning

> on P16 through P18. However, this hyperactivity did

> not reach statistical significance. In addition, this

> hyperactive behavior was blocked by pre-treatment

> with vitamin E (Fig. 3).

> GFAP and synaptophysin: Previous work in our lab

> revealed retarded neurobehavioral development in mice

> treated with 600 mg kg & #61485;1 VPA on embryonic day 13[1].

> In order to determine whether we could detect a

> Am. J. Biochem. & Biotech., 4 (2): 218-225, 2008

> 222

> Motor activity:

> Day of testing

> 13 14 15 16 17 18 19 20

> Number of horizontal beam breaks

> 0

> 200

> 400

> 600

> 800

> 1000

> Corn oil+saline

> Corn oil+VPA

> Vitamin E+VPA

> Fig. 3: Motor activity: Horizontal beam breaks for

> groups of pups treated with VPA (400 mg kg & #61485;1)

> or saline on P14. Some groups received vitamin

> E pretreatment while others received corn oil.

> *: p<0.05 compared to corn oil/saline

> Prenatal saline

> Sacrificed day 4

> Prenatal saline

> Sacrificed day 5

> Pprenatal VPA

> Sacrificed day 5

> Glia-fibrillary acidic protein

> 0

> 1000

> 2000

> 3000

> 4000

> 5000

> 6000

> 7000

> SALINE (PND4) SALINE (PND5) VPA (PND5)

> Protein level (arbitrary units)

> *

> Animals treated with VPA 600 mg kg & #61485;1 or Saline on E13

> Fig. 4: GFAP: Pups received either VPA (600 mg kg & #61485;1)

> or saline in utero on E13 and were sacrificed on

> either P4 or P5. *: p<0.05 compared to corn

> oil/saline

> biological marker of VPA-induced neurobehavioral

> retardation, we examined early postnatal levels of

> GFAP and synaptophysin in the cerebellum following

> in utero exposure to VPA. Mice treated with

> 600 mg kg & #61485;1 VPA on E13 and sacrificed on P5 showed

> decreased concentrations of GFAP in the cerebellum

> compared to saline treated controls sacrificed on both

> P4 and P5 (F (2, 8) = 12.3, p<0.01) (Fig. 4). Likewise,

> the concentration of synaptophysin was significantly

> decreased in the cerebellum of E13 VPA-treated mice

> compared to both P4 and P5 saline controls

> Prenatal saline

> Sacrificed day 4

> Prenatal saline

> Sacrificed day 5

> Prenatal VPA

> Sacrificed day 5

> 0

> 1000

> 2000

> 3000

> 4000

> 5000

> 6000

> 7000

> 8000

> SALINE (PND4) SALINE (PND5) VPA (PND5)

> Protein level (arbitrary units)

> *

> Synaptophysin

> Animals treated with VPA 600 mg kg & #61485;1 or Saline on

> E13

> Fig. 5: Synaptophysin: Pups received either VPA

> (600 mg kg & #61485;1) or saline in utero on E13 and

> were sacrificed on either P4 or P5. *: p<0.05

> compared to corn oil/saline

> (F (2, 6) = 10.0, p<0.01) (Fig. 5). During this period of

> postnatal development, GFAP and synaptophysin levels

> increase to promote normal astrocytes-neuron

> interactions and synaptogensis, respectively[37].

> Therefore, mice treated with VPA in utero show

> immature neural development, since the levels of GFAP

> and synaptophysin observed on P5 are much lower than

> those found in mice from an earlier postnatal period

> (P4). This suggests that the behavioral retardations seen

> in our previous study are influenced by retarded neural

> development.

> The etiology of autism is thought to involve early

> exposure to ROS-generating toxicants acting upon

> genetically-sensitive individuals. We have developed a

> new strategy to assess the detrimental effects of early

> toxicant exposure on neurobehavioral development,

> classifying the behavioral deficits as retardations,

> regressions or intrusions. In previous studies we

> demonstrated that early exposure to VPA or MeHg

> results in behavioral deficits in the maturation of social,

> cognitive and motor skills[1,8]. Furthermore, we

> demonstrated that our behavioral model was useful in

> demonstrating that pretreatment with an antioxidant

> protected mice against the behavioral deficits induced

> by early exposure to MeHg[8]. In the present study, we

> demonstrated that vitamin E was capable of protecting

> mice against VPA-induced regression in negative

> geotaxis and mid-air righting as well as against

> intrusive VPA-induced hyperactivity. Collectively, the

> present data together with our previous MeHg study

> indicate that the generation of ROS may be a common

> Am. J. Biochem. & Biotech., 4 (2): 218-225, 2008

> 223

> factor mediating toxicant-induced neuronal damage

> associated with autism and that neurobehavioral

> assessments provide an important functional measure of

> the potential benefits of antioxidants.

> A second objective of the present study was to

> develop a biological marker of the VPA-induced

> damage. Toward this end we used our initial model,

> delivering the VPA prenatally on E13[1]. We had

> demonstrated that this prenatal VPA treatment resulted

> in later behavioral deficits as assessed in the surface

> and mid-air righting tests, negative geotaxis and in

> water maze. Furthermore, this prenatal VPA treatment

> resulted in sex-dependent differences in these

> behavioral deficits with males more affected than

> females. In the present study, we found that both

> cerebellar GFAP and cerebellar synaptophysin were

> reduced postnatally following the prenatal VPA

> administration. GFAP is a marker of astroglia in the

> brain and is involved in astrocyte-neuron interactions.

> GFAP mutant mice have abnormal structure and exhibit

> deficient long-term depression in cerebellar Purkinje

> cell synapses[37]. Therefore, major alterations in GFAP

> may alter Purkinje cell communication that, in turn,

> may alter behavior. Synaptophysin is a widely used

> marker for nerve terminals and can indicate

> synaptogenesis. Therefore, a reduction in

> synaptophysin in the cerebellum could signify a

> reduction in synatpogenesis in that region. More

> generally, it is intriguing that these biological markers

> may reflect the behavioral deficits of cognitive and

> motor retardation caused by the early VPA exposure.

> Future studies are designed to determine if the

> antioxidant pretreatment also protects the mice against

> these neurological changes induced by the VPA.

> There is ample evidence that ROS are involved in

> human autism. Free oxygen radicals could result from

> ingested or inhaled environmental toxins, food or food

> additives, inflammation or infection (overt or occult).

> The interaction of free oxygen radicals and

> polymorphic oxidative genes during gestation or

> postnatally could disrupt neurogenesis in developing

> brain at multiple time windows, eliciting immediate

> stage-dependent effects in specific systems that

> influence subsequent ontogenetic processes, leading to

> the phenotype of autism. Indeed an exacerbated

> oxidative stress response has been implicated in autism.

> Specifically lower glutathione peroxidase (GPX) and

> superoxide dismutase (SOD) activity were found in

> children with autism[38-40]. An increase in body burden

> of various toxins was reported in autism[41,42]. In

> addition, provoked urinary mercury excretion is found

> to be higher in autism[43]. These toxins could generate

> oxidative stress in children with autism. Elevated nitrite

> and nitrate in plasma[44] and red cells[38] have been

> reported in children with autism. This elevation

> indicates excess generation of nitric oxide free radicals.

> In addition, two independent double blind placebo

> controlled clinical trials of antioxidants (vitamin C or

> carnosine) showed beneficial effects in autism[45,46].

> Finally, we conducted a study of oxidative stress

> biomarkers in children with autism and age matched

> healthy controls. Our results showed that urinary

> excretion of 8 isoprostane F2_ was significantly higher

> in children with autism as compared to healthy

> controls[47]. There was also a trend of increased 8-

> OHdG urinary excretion in autistic subjects. These

> results suggest that oxidative stress is exacerbated in

> autism and are consistent with the present results of

> antioxidant protective effects against VPA-induced

> behavioral deficits in mice.

> In summary, we have developed a comprehensive

> neurobehavioral model in which mice are exposed to

> candidate toxicants during critical periods of neural

> development. The mice may have altered expression of

> genes thought to be associated with autism and/or to

> confer increased sensitivity to the toxicants. The mice

> are then assessed in a battery of tests designed to assess

> behavioral maturation of skills in the social, cognitive

> and motor domains. Toxicant or genetic-induced

> deficits in the behavioral maturation are classified as

> retardations, regressions or intrusions. In the present

> studies, we further demonstrate that pretreatment with

> an antioxidant protects the mice against the toxicantinduced

> behavioral deficits. We conclude that our

> model is useful for evaluation of the theory that

> oxidative stress may play a role in the etiology of

> autism.

> ACKNOWLEDGEMENT

> Supported by ES11256, EPAR829391, New Jersey

> Governor's Council on Autism and Autism Speaks.

> REFERENCES

> 1. Wagner, G.C., K.R. Reuhl, M. Cheh, M., P. McRae

> and A.K. Halladay, 2006. A new neurobehavioral

> model of autism in mice: Pre- and Postnatal

> exposure to sodium valproate. J. Autism Dev. Dis.,

> 36: 779-793.

> 2. Bernard, S., A. Enayati, L. Redwood, H. and

> T. Binstock, 2000. Autism: A novel form of

> mercury poisoning. ARC Res., 1-12.

> 3. Dawson, G., 1996. Neuropsychology of autism: a

> report on the state of the science. J. Aut. Dev. Dis.,

> 26: 179.

> Am. J. Biochem. & Biotech., 4 (2): 218-225, 2008

> 224

> 4. Chauhan, A. and V. Chauhan, 2006. Oxidative

> stress in autism. Pathophysiol., 13: 171-181.

> 5. Chauhan, A., V. Chauhan, T. Brown and I. Cohen,

> 2004. Oxidative stress in autism: increased lipid

> peroxidation and reduced serum levels of

> ceruloplasmin and transferin-the antioxidant

> proteins. Life Sci., 75: 2539-2549.

> 6. , S.J., P. Cutler, S. Melnyk, S. Jernigan,

> L. Janak, D.W. Gaylor and J.A. Neubrander, 2004.

> Metabolic biomarkers of increased oxidative stress

> and impaired methylation capacity in children with

> autism. Am. J. Clin. Nutr., 80: 1611-1617.

> 7. Wagner, G.C., K.R. Reuhl, X. Ming and

> A.K. Halladay, 2007. Behavioral and

> neurochemical sensitization to amphetamine

> following early postnatal administration of

> methylmercury. NeuroToxicol., 281: 59-66.

> 8. Cheh, M.A., A.K. Halladay, K.R. Reuhl,

> M. Polunas, X. Ming and G.C. Wagner, 2007.

> Trolox, a Vitamin E derivative, protects against

> persistent neurobehavioral disruption induced by

> neonatal methylmercury exposure. Soc. Toxicol.,

> NC.

> 9. Rodier, P.M., J.L. Ingram, B. Tisdale, S.

> and J. Romano, 1996. Embryological origin for

> autism: developmental abnormalities of the cranial

> nerve motor nuclei. J. Comp. Neurol.,

> 370:247-261.

> 10. Ingram, J.L., S.M. Peckham, B. Tisdale and

> P.M. Rodier, 2000a. Prenatal exposure to valproic

> acid reproduces the cerebellar anomalies associated

> with autism. Neurotox. Teratol., 22, 319-324.

> 11. Sobaniec-Lotoweska, M.E., 2001. Ultrastructure of

> purkinje cell perikara and their dendritic processes

> in the rat cerebellar cortex in experimental

> encephalopathy induced by chronic application of

> valproate. Int. J. Exp. Pathol., 82: 337-348.

> 12. , S.J., P. Turnpenny, A. Quinn, S. Glover

> and D.J. Lloyd, 2000. Montgomery and

> J.C.S. Dean, A clinical study of 57 children with

> fetal anticonvulsant syndromes. J. Med. Genetics,

> 37: 489-497.

> 13. , G., J. King, M. Cunningham,

> M. Stephan, B. Kerr and J.H. Hersh, 2001. Fetal

> valproate syndrome and autism: additional

> evidence of an association. Dev. Med. Child

> Neurol., 43: 202-206.

> 14. Chapman, J.B. and M.G. Cutler, 1989. Effects of

> sodium valproate on development and social

> behaviour in the Mongolian gerbil. Neurotoxicol.

> Teratol., 11: 193-198.

> 15. Voorhees, C.V., 1987. Behavioral teratogenicity of

> valproic acid: selective effects on behavior after

> prenatal exposure to rats. Psychopharmacol.,

> 92: 173-179.

> 16. Schneider, T. and R. Przewlocki, 2005. Behavioral

> alterations in rats prenatally exposed to valproci

> acid: Animal model of autism.

> Neuropsychopharmacol., 30: 80-89.

> 17. Altman, J. and S.A. Bayer, 1978. Prenatal

> development of the cerebellar system in the rat.

> Cytogenesis and histogenesis of the deep nuclei

> and cortex of the cerebellum. J. Comp. Neurol.,

> 179: 23-48.

> 18. Inouye, M. and U. Murakami, 1980. Temporal and

> spatial patterns of purkinje cell formation in the

> mouse cerebellum. J. Comp. Neurol.,

> 194: 499-503.

> 19. Bachevalier, J. and M. Beauregard, 1993.

> Maturation of medial temporal lobe memory

> functions in rodents, monkeys and humans.

> Hippocampus, 3:191-202.

> 20. Rice, D. and S. Barone, 2000. Critical periods of

> vulnerability for the developing nervous system:

> Evidence from human and animal models. Environ.

> Health Per., 108: 511-533.

> 21. Murugesan, V. and P. Subramanian, 2003.

> Enhancement of circulatory antioxidants by & #61537;ketoglutarate

> during sodium valproate treatment in

> wistar rats. Polish J. Pharmacol., 55: 31-36.

> 22. Tong, V., T.K.H. Chang, J. Chen and F.S. Abbott,

> 2003. The effect of valproic acid on hepatic and

> plasma levels of 15-F2t-Isoprostane in rats. Free

> Radical Biol. Med. 34: 1435-1446.

> 23. Sobaniec-Lotowska, M.E., 1997. Effects of longterm

> administration of the antiepileptic drugsodium

> valproate upon the ultrastructure of

> hepatocytes in rats. Exp. Toxicol. Pathol.,

> 49: 225-32.

> 24. Myers, G.J. and P.W. son, 2000. Does

> methylmercury have a role in causing

> developmental disabilities in children? Environ.

> Health Per., 108: 413-420.

> 25. Rice, D.C., 1996. Sensory and cognitive effects of

> developmental methylmercury exposure in

> monkeys and a comparison to effects in rodents.

> Neurotoxicol., 17, 139-154.

> 26. Dey, P.M. and K.R. Reuhl, 1999. Developmental

> methylmercury administration alters cerebellar

> PSA-NCAM expression and Golgi sialyltransferase

> activity. Res., 845:139-151.

> 27. Yee, S. and B.H. Choi, 1996. Oxidative stress in

> neurotoxic effects of methylmercury poisoning.

> Neurotoxicol., 17: 17.

> Am. J. Biochem. & Biotech., 4 (2): 218-225, 2008

> 225

> 28. Park, S.T., K.T. Lim, Y.T. Chung and S.U. Kim,

> 1996. Methylmercury-induced neurotoxicity in

> cerebral neuron culture is blocked by antioxidants

> and NMDA receptor antagonists. Neurotoxicol.,

> 17: 37.

> 29. Usuki, F., A. Yasutake, F. Umehara, H. Tokunaga

> and M. Matsumoto et al., 2001. In vivo protection

> of a water-soluble derivative of vitamin E, Trolox,

> against methylmercury-intoxication in the rat

> Neurosci Lett., 304: 199-203.

> 30. Sanfeliu, C., J. Sebastia and S.U. Ki, 2001.

> Methylmercury neurotoxicity in cultures of human

> neurons, astrocytes, neuroblastoma cells.

> Neurotoxicol., 22: 317-327.

> 31. Aschner, M., C.P. Yao, J.W. and K.H. Tan,

> 2000. Methylmercury alters glutamate transport in

> astrocytes. Neurochem. Int., 37: 19.

> 32. Stohs, S.J. and D. Bagchi, 1995. Oxidative

> mechanisms in the toxicity of metal ions. Free

> Radical Biol. Med., 18: 321.

> 33. Sorg, O., B. Schilter, P. Honegger and

> F. Monnet-Tschudi, 1998. Increased vulnerability

> of neurones and glial cells to low concentrations of

> methylmercury in a prooxidant situation. Acta

> Neuropathol. (Berl), 96: 621.

> 34. Mundy, W.R. and T.M. Freudenrich, 2000.

> Sensitivity of immature neurons in culture to

> metal-induced changes in reactive oxygen species

> and intracellular free calcium. Neurotoxicol,

> 21: 1135.

> 35. Altman, J. and K. Sudarshan, 1975. Postnatal

> development of locomotion in the laboratory rat.

> Animal Behav., 23: 896-901.

> 36. Petrosini, L., M. Molinari and T. Gremoli, 1990.

> Hermicerebellectomy and motor behavior in rats.

> II. Effects of cerebellar lesion performed at

> different developmental stages. Exp. Brain Res.,

> 82: 483-492.

> 37. Shibuki, K., H. Gomi, L. Chen, S. Bao, J.J. Kim,

> H. Wakatsuki, T. Fujisaki, K. Fujimoto, A. Katoh,

> T. Ikeda, C. Chen, R.F. and S. Itohara,

> 1996. Deficient cerebellar Long-term depression,

> impaired eyeblink conditioning and normal motor

> coordination in GFAP mutant mice. Neuron,

> 16: 587-599.

> 38. Sogut, S., S.S. Zoroglu and H. Ozyurt et al., 2003.

> Changes in nitric oxide levels and antioxidant

> enzyme activities may have a role in the

> pathophysiological mechanisms involved in

> autism. Clinica Chimica Acta, 331: 111-117.

> 39. Golse, B., P. Debray-Ritzen and P. Durosay et al.,

> 1978. Perturbation de deux enzymes, la

> superoxyde-dismutase I et la glutathioneperoxydase

> dans la psychose infantile de

> developpement (autisme infantile). Rev. Neurol.

> (Paris), 134: 699-705.

> 40. Yorbik, O., A. Sayal and C. Akay et al., 2002.

> Investigation of antioxidant enzymes in children

> with autistic disorder. Prostaglandins Leukot

> Essent Fatty Acids, 67: 341-343.

> 41. Edelson, S.B. and D.S. Cantor, 1998. Autism:

> xenobiotic influences. Toxicol. Ind. Health, 14:

> 799-811.

> 42. Edelson S.B. and D.S. Cantor, 2000. The

> neurotoxic etiology of the autistic spectrum

> disorder: a replicative study. Toxicol. Ind. Health,

> 16: 239-47.

> 43. Bradstreet, J., D.A. Geier and J.J. Kartzinel et al.,

> 2003. A case-control study of mercury burden in

> children with autistic spectrum disorders. J. Am.

> Phy. Sur., 8: 76-79.

> 44. Zoroglu, S.S., M. Yurekli and I. Meram et al.,

> 2003. Pathophysiological role of nitric oxide and

> adrenomedullin in autism. Cell Biochem. Function,

> 21: 55-60.

> 45. Chez, M.D., C.P. Buchanan and M.C. Aimonovitch

> et al., 2002. Double-blind, placebo-controlled

> study of l-carnosine supplementation in children

> with autistic spectrum disorders. J. Child Neurol.,

> 17: 833-837.

> 46. Dolske, M.C., J. Spollen and S. McKay et al.,

> 1993. A preliminary trial of ascorbic acid as

> supplemental therapy for autism. Prog. Neuro-

> Psychopharm. Biol. Psychiat., 17: 765-774.

> 47. Ming, X., P. Stein, M. Brimacombe, W. ,

> G. Lambert and G.C. Wagner, 2005. Increased

> excretion of a lipid peroxidation biomarker in

> autism. Prostaglandins, Leukotrienes and Essential

> Fatty Acids, 73: 379-384.

> ~~~~~~~~~~~

>

> =====

>

February 23, 2008

From the study I just posted:

" Nonetheless, it

is important to note that autism was not found to be

associated with either pre- or neonatal exposure to

organic mercury. "

=====

February 23, 2008

I am the Carolyn referred to in this post as taking my daughter to Dr. Ming

who was mentioned in the research/report.

I am getting confused. Just in case others are getting confused, let me give

you some background because I think cutting and pasting old emails may be

difficult to understand the point trying to be made (which I admit I don't

understand right now).

My daughter went to Dr. Ming a couple of times, the last time being 2004.

She didn't mention cheek samplings. The only test she did on my daughter was a

repeat brain MRI.

We did discuss proefa. I was giving my daughter efelex prior to age 3 and

then switched to proefa after reading the posts on this list from so many saying

they saw progress in speech in their children after supplementing with it.

The efelex did cause positive behavorial/sensory changes. My daughter had been

grinding her teeth all day (not at night). A doctor from NY City (Dr.

Pescatore) suggested efa supplementation for this. Within 3 weeks, to my

surprise

and delight, the grinding stopped. Then, when we started the proefa, it was

like a miracle because my daughter started to say her first purposeful " mama "

and other words. That was at about 3 and 1/2. The problem for us was that

unlike many others, the progress leveled off but sometimes we would see some

progress when stopping and starting the proefa again.

In our case, diet has also been important because at around age 5, my

daughter started to have seizures (convulsions) when she had a sudden

temperature

increase. Dr. Ming suggested our seeing a different neurologist when the

seizures started. One doctor suggested a modified atkins diet, and it appears

(please God) that the seizures have stopped. She has not had one in a year. So,

basically, she eats fruits, veggies, whole grains, fish, meat, and soups. All

around, her health has improved too.

Now, the first MRI was done when she was 2 and 1/2 as ordered by Dr. Arnold

Gold. The same radiologist/neur did this MRI and the later one. It was

determined that her myelin had not all come in but it was minimal. Dr's do

interpret MRI's differently though because when we were at Children's Hospital

in

Philadelphia to make sure there was no genetic reason for her delays (there

were

none), our geneticist showed the first MRI to a neurologist who asked if it

was the brain of a premature child (my daughter was 4 wks premature but also I

had labor pains at 8 wks early). The second MRI showed that the myelin was

completely in (according to radiologist/neuro-Dr. Ming thought maybe a bit

more needed) but now they could see tiny cysts in the white matter caused by an

apparent bleed either prior to birth or during the traumatic birth.

The white matter, I was told deals with processing issues, and Dr. Ming was

happy that her gray matter was not affected at all since that would affect

intelligence.

Ok, so I hope this helps anyone interested.

Also, I know no one would ever mean to offend anyone, but I would appreciate

it if the term, " mentally retarded " or " mental retardation " was not used.

More and more doctors and even governmental agencies are realizing that it has

a negative connotation to it and has been used in insulting ways. The terms

that are used more and more are developmentally disabled or cognitively

impaired. Thanks for your consideration on this. Carolyn

**************Ideas to please picky eaters. Watch video on AOL Living.

(http://living.aol.com/video/how-to-please-your-picky-eater/rachel-campos-duffy/

2050827?NCID=aolcmp00300000002598)

February 23, 2008

So E only works on afterbirth exposure?

>

> From the study I just posted:

>

> " Nonetheless, it

>

> is important to note that autism was not found to be

>

> associated with either pre- or neonatal exposure to

>

> organic mercury. "

>

> =====

>

February 23, 2008

Forgot to mention in my long email, that Dr. Ming had told me at the time

that she thought that the improvement shown in the second MRI that showed some

cysts had disappeared or greatly decreased, was caused by the vitamin E found

in the efa supplementation I had been giving her (efelex and then proefa).

Carolyn

**************Ideas to please picky eaters. Watch video on AOL Living.

(http://living.aol.com/video/how-to-please-your-picky-eater/rachel-campos-duffy/

2050827?NCID=aolcmp00300000002598)

February 23, 2008

No. That's not how I read it. The portion quoted to you below

appears to be taken out of context (and incomplete in it's suggested

response to your observation.) Reading the entire article, the

researchers do not appear to intend to be so limiting their study

and/or their hypotheses, opinions, and predictions.

For instance, also from the study:

" In previous studies we have demonstrated a sensitization response

to dopaminergic toxicants in adult mice following prenatal

administration of MeHg[7]. We have also demonstrated that

antioxidant pretreatment protects mice against the behavioral

deficits induced by early exposure to MeHg[8]. "

This quote follows their summary hypothesis: " ...that autism may be

the result multiple exposures to any of a number of toxicants; the

initial exposure sensitizes the subject such that later exposures to

the same or different toxicants results in an enhanced oxidative

stress response. ... "

Anywho.... :-)

> >

> > From the study I just posted:

> >

> > " Nonetheless, it

> >

> > is important to note that autism was not found to be

> >

> > associated with either pre- or neonatal exposure to

> >

> > organic mercury. "

> >

> > =====

> >

February 23, 2008

kiddietalk wrote:

> From the study I just posted:

> " Nonetheless, it is important to note that autism was not found to be

> associated with either pre- or neonatal exposure to

> organic mercury. "

Included in the cited report or not - this is NOT a true statement....no

100% conclusive causal link has been found NOR HAS IT BEEN DISPROVED at

this point by anybody that organic mercury (Thimerosal) ALONE triggers

or doesn't trigger autism/autoimmune problems...but studies are still in

progress for both sides of the discussion...and the " causes " are not

focused on a single environmental contaminant (i.e. organic

mercury/thimerosal)...there is agreement that it is a NEUROtoxin and

what damage a NEUROtoxin does to the head of the NEUROlogical system is

pretty obvious...it kills NEUROns and does create a damage to the

NEUROlogical system. It is only one of many toxins in pre- or neonatal

exposures that contributes to lots of health issues.

What gets said publicly and by who depends on who's reporting it and who

is getting paid by or receiving research grants or advertising monies

from the very pharmaceutical companies who continue to produce vaccines

with thimerosal (organic mercury)...leaving even traces (while

officially not using it labeled as a " preservative " but it is still used

during the production phase and " filtered " out leaving levels which are

still 1000 times more than the EPAs minimum allowed ppm (parts per

million) for drinking water. (anyone new who wants to know more can

check out the group EOHarm but warning that it is much more high

volume than this apraxia group). It's just ONE facet of how

environmental toxins in general are messing with humans.

If the door is opened to discuss this subject - be prepared before

presenting it as a " fact " especially if you have not yet read up on ALL

the research on both sides currently out there. I'm sure other biomed

moms will reply publically to this...so that all the other members don't

get misled. Those of us who've been researching all of this stuff

for our own kids for years will beg to differ based on validity of the

statement considering the 100's of scientific research reports

(including published ones in scientific journals) and ongoing

investigative reports we've made time to locate and read in their

entirety...oh and the documented medical problems, including apraxia,

that our kids are forced to overcome.

What you pointed out, truth or mistruth, is a subject much larger and

controversial than you may have possibly imagined and

is probably not the best venue to continue it, but

will probably become yet another " hot topic " while things get clarified

yet again for the new families coming into the discussion. " Hot

topics " seem to be the trend this weekend already though.

____________________________________________________________

Receive Notifications of Incoming Messages

Easily monitor multiple email accounts & access them with a click.

Visit http://www.inbox.com/notifier and check it out!

February 23, 2008

That is what I thought. My son, while not autistic (yet anyway but he

shows no signs now but once did) was exposed to mercury in utero at a

dental appt. Were he not a bleeder I'd be jumping on E. Let's hope

the dietary E remains enough.

> > >

> > > From the study I just posted:

> > >

> > > " Nonetheless, it

> > >

> > > is important to note that autism was not found to be

> > >

> > > associated with either pre- or neonatal exposure to

> > >

> > > organic mercury. "

> > >

> > > =====

> > >

February 23, 2008

In a message dated 2/23/2008 7:04:03 P.M. Eastern Standard Time,

crystalam_@... writes:

(I figured she could have passed it to me).

Anyway, turns out my Uranium (from Radon probably) was off the charts,

and my daughters was 1 1/2 times mine. So you might be right, and it

might not be mercury, but that doesn't mean it's not something else!

It seems like different things are the triggers in different cases,

but if you look it seems like it's usually some sort of toxin built

up. Just my .02

Again-- In my case-- my toxin levels were fine. No uranium problems etc.

I am TOTALLY hearing what you're saying though, and AGREE that many times

the cause of these neurological based disabilities are exposures to metals and

toxins. (but will say definitely NOT ALL)

becky

**************Ideas to please picky eaters. Watch video on AOL Living.

(http://living.aol.com/video/how-to-please-your-picky-eater/rachel-campos-duffy/

2050827?NCID=aolcmp00300000002598)

February 23, 2008

Also, fwiw, it is not clear to me that this study attempts to assert

that Trolox (a water soluble Vit. E) remediates prior effects or

harm, or whether it only acts as a pretreatment prophylactic measure

before the toxic exposure in question. But, I'm also pretty sure

I'm probably oversimplifying the analysis! Because, although I

understand we encourage fish oil and Vitamin E protocols, I'm not

sure I've ever known whether the goal is to reverse prior or prevent

future damage; or, for that matter, whether oxidative stress itself

causes some of the issues presented. These questions I'll leave to

the researching MDs. :-)

> > >

> > > From the study I just posted:

> > >

> > > " Nonetheless, it

> > >

> > > is important to note that autism was not found to be

> > >

> > > associated with either pre- or neonatal exposure to

> > >

> > > organic mercury. "

> > >

> > > =====

> > >

February 23, 2008

In a message dated 2/23/2008 7:18:58 P.M. Eastern Standard Time,

lizlaw@... writes:

As for mercury exposure...depends what you mean.

If you have mercury amalgams, went to the dentist, live in NJ (LOL)

there was likely some exposure.

You're probably thinking with THIS current pregnancy-- and not Asa's. ????

With THIS pregnancy-- I've been to the dentist and the mercury fillings on

one side taken out, so the concern would be present NOW for THIS pregnancy.

With ASA though, who is my Apraxic child--- there definitely wasn't any of

these issues.

Becky

**************Ideas to please picky eaters. Watch video on AOL Living.

(http://living.aol.com/video/how-to-please-your-picky-eater/rachel-campos-duffy/

2050827?NCID=aolcmp00300000002598)

February 23, 2008

Please excuse my ignorance here, but we're extrapolating quite a bit

to suggest a correlation between this study and our use of Vit. E,

in its natural form, in apraxic kids, right? I ask now only because

I'm just focusing on the fact that this study is using a " brand

named " water soluble derivative of Vit. E that currently appears

available for research purposes.

Don't the archives recommend the fat soluble Vit. E in it's natural

form (d-)?

Anyway, I'm pretty happy to see that studies like this are underway

regardless. I would love to know the material differences in the

Vit E's and whether this research can be effectively applied in our

context using natural Vit. E or whether that may be a reach.

>

> American Journal of Biochemistry and Biotechnology 4 (2): 218-225,

> 2008

> ISSN 1553-3468

> Corresponding Author: C. Wagner, Psychology, Busch Campus,

> Rutgers University, New Brunswick, NJ 08854

> Tel: 732-445-4660 Fax: 732-445-2263

> 218

> Evidence of Oxidative Stress in Autism Derived from Animal Models

> 1Xue Ming, 2 A. Cheh and 2 L. Yochum,

> 3Alycia K. Halladay and 2 C. Wagner

> 1Pediatric Neuroscience, UMDNJ, Newark, NJ

> 2Psychology, Rutgers University, New Brunswick, NJ

> 3Autism Speaks, Princeton, NJ

> Abstract: Autism is a pervasive neurodevelopmental disorder that

> leads to deficits in social

> interaction, communication and restricted, repetitive motor

> movements. Autism is a highly heritable

> disorder, however, there is mounting evidence to suggest that

> toxicant-induced oxidative stress may

> play a role. The focus of this article will be to review our

animal

> model of autism and discuss our

> evidence that oxidative stress may be a common underlying

mechanism

> of neurodevelopmental

> damage. We have shown that mice exposed to either methylmercury

> (MeHg) or valproic acid (VPA) in

> early postnatal life display aberrant social, cognitive and motor

> behavior. Interestingly, early exposure

> to both compounds has been clinically implicated in the

development

> of autism. We recently found

> that Trolox, a water-soluble vitamin E derivative, is capable of

> attenuating a number of

> neurobehavioral alterations observed in mice postnatally exposed

to

> MeHg. In addition, a number of

> other investigators have shown that oxidative stress plays a role

in

> neural injury following MeHg

> exposure both in vitro and in vivo. New data presented here will

show

> that VPA-induced

> neurobehavioral deficits are attenuated by vitamin E as well and

that

> the level of glial fibrillary acidic

> protein (GFAP), a marker of astrocytic neural injury, is altered

> following VPA exposure. Collectively,

> these data indicate that vitamin E and its derivative are capable

of

> protecting against neurobehavioral

> deficits induced by both MeHg and VPA. This antioxidant protection

> suggests that oxidative stress

> may be a common mechanism of injury leading to aberrant behavior

in

> both our animal model as well

> as in the human disease state.

> Key words: Vitamin E, trolox, valproic acid

> INTRODUCTION

> The core symptoms of autism include language

> deficits, impaired social interactions and inappropriate,

> stereotypic and sometimes self-injurious behaviors. The

> etiology of autism remains unknown but may involve

> early exposure to environmental toxicants acting upon

> genetically-sensitive individuals. No single toxicant has

> been identified; rather a broad range of toxicants

> including drugs, metals, solvents, herbicides, pesticides,

> etc. have been associated with autism[1-3]. A common

> feature across this range of potential compounds is

> toxicant-induced oxidative stress causing neuronal

> damage leading to the behavioral phenotype of

> autism[4-6]. Likewise, no single gene has been identified

> but, rather, a constellation of as many as 15

> polymorphisms may ultimately predispose the

> individual to autism. Again, genetic alterations leading

> to compromised handling of toxicant-induced reactive

> oxygen species (ROS) has been a common theme.

> Since the etiology of autism is unknown, it is

> essential that animal models be developed. The

> behavioral symptoms of autism have proven difficult to

> model in other species. Accordingly, we have initiated

> work on a novel strategy to model the behavioral

> phenotype of autism in mice[1]. In this model, the

> normal development of key behaviors is carefully

> monitored from birth through adolescence. Once the

> maturation of these key behaviors is understood in

> terms of the postnatal day(s) of life in which subjects

> are able to successfully perform the task or engage in

> the behavior, the performance of mice with early

> toxicant exposure and/or genetic modification can be

> assessed.

> The model strategy begins by characterizing

> behavioral manifestations of developmental disorders

> as retardations (a behavior fails to develop during a

> critical period of maturation), regressions (a behavior

> develops at about the right time but then is lost with

> Am. J. Biochem. & Biotech., 4 (2): 218-225, 2008

> 219

> later development, especially following toxicant

> exposure), or intrusions (the appearance of behaviors

> aberrant in form or frequency which mask normal

> development). Most developmental disorders include

> some combination of these conditions. In this

> framework, the hypothesis that environmental toxicants

> or genetic alterations are causally involved in autism

> can be readily tested. That is, acute or repeated

> exposure to a toxicant should disrupt neurobehavioral

> development causing behavioral retardation, regression,

> or intrusions and these toxicant-induced behavioral

> deficits should occur at lower doses in the geneticallysensitive

> mice. Traditionally, animal models of

> developmental disorders have not examined these three

> scenarios of retardations, regressions and/or intrusions

> but, instead, focus on single aspects of neurobehavioral

> development. The judicious use of toxicants associated

> with autism or toxicants known to damage brain regions

> associated with autism confers some selectivity of the

> model for autism. Likewise, manipulation of genes

> associated with autism also confers some selectivity of

> this model for autism. Finally, administering a battery

> of tests that assess social, cognitive and motor

> maturation of the mice confers some selectivity for

> autism. Ultimately, it is the possibility of combining

> select toxicant exposure in genetically-sensitive mice

> followed by thorough assessment of social, cognitive

> and motor skill maturation that makes this a

> comprehensive animal model of autism.

> In our initial studies, we identified toxicant induced

> retardation of motor and cognitive skills following pre-

> or post-natal exposure to sodium valproate (VPA).

> Likewise, we were able to demonstrate dramatic loss of

> acquired skills, i.e. regressions, following post-natal

> VPA administration[1]. Finally, we demonstrated

> toxicant induced intrusions wherein toxicant-treated

> subjects exhibited dramatic increases in stereotypic and

> self-injurious behaviors akin to those seen in autism[7,8].

> VPA was chosen as our first agent to test this

> model following reports of an association between

> autism and prenatal exposure to this teratogen[9-13].

> Previous studies have also demonstrated impairment in

> cognitive, motor, attention and social development in

> rats administered pre- or post-natal VPA[14-17].

> Accordingly, in our first studies[1] mice were exposed to

> VPA either in utero or post-natally. The prenatal

> exposure time reflected a period of cerebellar Purkinje

> cell generation differentiation in the mouse[10,14,17,18].

> The post-natal time of P14 was based on our

> observation that critical cerebellar-mediated behaviors

> of mid-air righting and negative geotaxis mature or first

> appear on this day in the mouse[1] and because of

> continued neuronal and glial development in other brain

> regions[15,19,20]. Of importance, VPA administration

> results in high levels of markers for oxidative stress and

> lipid peroxidation including 15-F-isoprostane and

> thiobarbituric acid reactive substances[21-23].

> An organic mercury, MeHg, was selected as our

> second compound for testing because it is an important,

> widely distributed environmental toxicant. MeHg does

> cross the placental barrier and, in humans exposed in

> utero to acute high doses, was shown to cause

> retardation in cognitive and locomotor development

> along with numerous other neurological symptoms

> including seizures and cerebral palsy[24]. Nonetheless, it

> is important to note that autism was not found to be

> associated with either pre- or neonatal exposure to

> organic mercury.

> The consequences of low dose, chronic exposure to

> mercury through fish consumption are somewhat more

> controversial with some studies showing deleterious

> effects while others show no adverse consequences[2,24].

> Early exposure to mercury has been shown to disrupt

> the neurobehavioral development of other species

> including rodents and primates[25]. The mechanism

> through which MeHg exerts its toxicity is thought to be,

> in part, mediated by disruption of neural cell adhesion

> molecules[26]. In addition, oxidative stress is involved in

> MeHg-induced neurotoxicity as demonstrated by

> increased ROS and thiobarbituric acid reactive

> substances and a reduction in GSH levels[27]. In

> addition, the neurotoxicity of MeHg in cultured neurons

> was blocked by the pretreatment with antioxidants[28].

> Trolox, a water-soluble derivative of vitamin E,

> protects against MeHg-induced neurotoxicity in rats[29].

> Likewise, antioxidants produced protective effects

> against MeHg toxicity in cultured human neurons and

> astrocytes[30]. Indeed, ROS have been implicated in

> MeHg-induced neurotoxicity in multiple experimental

> models[27,31-34]. Finally, we have recently demonstrated

> that pretreatment with Trolox protects mice against the

> neurobehavioral deficits induced by postnatal MeHg[8].

> Collectively, these data indicate that early exposure to

> MeHg causes neurobehavioral deficits consequent, at

> least in part, to the generation of ROS.

> In summary, wide ranges of toxicants and genetic

> alterations have been associated with autism. The

> toxicants are thought to have a common mechanism of

> generating ROS[4-6] while the genetic alterations are

> thought to result in enhanced sensitivity to the

> deleterious effects of ROS. Accordingly, we now

> hypothesize that autism may be the result multiple

> exposures to any of a number of toxicants; the initial

> exposure sensitizes the subject such that later exposures

> to the same or different toxicants results in an enhanced

> Am. J. Biochem. & Biotech., 4 (2): 218-225, 2008

> 220

> oxidative stress response. Furthermore, we predict that

> this sensitization will be exacerbated in individuals with

> genetic alterations affecting their handling of ROS. In

> previous studies we have demonstrated a sensitization

> response to dopaminergic toxicants in adult mice

> following prenatal administration of MeHg[7]. We have

> also demonstrated that antioxidant pretreatment protects

> mice against the behavioral deficits induced by early

> exposure to MeHg[8]. Accordingly, the objective of this

> study was to determine if antioxidants administered as a

> pretreatment to VPA would protect the mice against the

> VPA-induced behavioral regression. In addition, we

> sought to determine if the early VPA administration

> would alter levels of glial fibrillary acidic protein

> (GFAP), a marker of astrocytic neural injury, thus

> serving as a biological marker for the VPA-induced

> behavioral deficits.

> MATERIALS AND METHODS

> Subjects: Male and female BALB/c mice (Taconic,

> Germantown, NY) were housed together in plastic

> cages with standard wood chip bedding and free access

> to food and water. All mice were maintained in an

> AAALAC-accredited facility under guidelines set forth

> by the National Institutes of Health. Lights were set on

> a 12 h on: 12 h off cycle and temperature was

> maintained at 25 & #61616;C. Females were checked before 10

> AM for presence of a vaginal plug which was recorded

> as day 0 of embryonic development. Day of birth was

> recorded as day 0 and all pups were labeled for

> individual identification. Body weight was measured

> daily. Female pups were removed from the cage on day

> 5. For the behavioral studies, the sodium valproate

> (Sigma) dose was 400 mg kg & #61485;1 with a saline vehicle and

> the vitamin E dose was also 400 mg kg & #61485;1 but with a

> corn oil vehicle. All injections were s.c. in a volume of

> 1.0 mL 100 & #61485;1 g body weight.

> Negative geotaxis: Negative geotropism was tested on

> postnatal days P13-19 by placing the mouse facing

> downward along a 45 & #61616;C incline. Latency to turn 180 & #61616;C

> such that the head was facing upward along the incline

> was recorded with a maximum of 30 seconds for each

> trial.

> Motor Activity: Motor activity was assessed on days

> P14-19. The chamber consisted of a black

> 42 & #61605;22 & #61605;14 cm Plexiglass box. Six infrared sensors

> placed approximately 7 cm apart and 2.5 cm above the

> floor were used to measure activity over a 10 min

> period.

> Mid-air righting: When a mouse is dropped upside

> down from a height of 45 cm onto a padded surface it

> engages in a mid-air righting reflex with orderly, rostrocaudal

> movements, initiated with head and concluded

> with the hindlimbs such that the animal lands on its

> paws. The behavior first appears on P13 and is fully

> achieved by P17[35]. Mid-air righting has been linked to

> cerebellum development[36]. For the mid-air righting

> test, mice were elevated 45 cm above a foam pad,

> dorsal side down. The animal was released and ability

> to right in mid air assessed scored as the mouse landing

> on its paws on two out of three trials each day. Mice

> were tested on P13-20.

> Protein determination: In order to determine changes

> in protein expression following VPA treatment at

> behaviorally significant time points, animals were

> treated with VPA 600 mg kg & #61485;1 or saline on E13[1] and

> assayed on days P4 and P5 with the cerebellum

> removed and stored at -70 & #61616;C. Protein analysis via gel

> electrophoresis, western blot and densiometry was

> performed according to the methods of Dey et al.[26]

> with some small variations. In summary, whole

> homogenate fractions were homogenized in 1:10 w/v of

> a Tris extraction buffer [50 mM TrisHCl, pH 7.4,

> 0.32M sucrose, 1 mM EDTA, 1 vial to 100 ml protease

> inhibitor (Sigma, St. Louis, MO). The supernatant was

> removed following centrifugation for 10 min at 1000xG

> and combined with equilibration buffer [0.125M

> TrisHCl, pH 6.8, 4% SDS, 20% glycerol, 10%

> mercapoethanol) and immediately heated for 30 min at

> 70 & #61616;C. Protein values were determined using the BCA

> protein assay (Pierce, Rockford, IL) modified for a

> BOBAS FARA II enzyme analyzer (Roche

> Diagnostics, Nutley, NJ). Samples of 10 & #61549;g protein

> were separated by SDS-PAGE on a 10%

> polyacrylamide gradient gel using a Bio-Rad Mini-

> Protean II System (Bio-Rad, Mellville, NY) for GFAP

> and synaptophysin. Proteins were transferred to

> nitrocellulose membranes and were washed twice for

> 10 min each in phosphate buffered saline (PBS) and

> blocked with 5% non-fat dry milk in PBS for 1 h prior

> to application of primary antibody. Immunoblotting for

> GFAP and synaptophysin was performed overnight at

> 4 & #61616;C. All primary antibodies were obtained from Fisher

> (Springfield, NJ). Anitgens were visualized following

> 1 h incubation with secondary peroxidase antibodies

> (Southern Biotechnology Associates, Birmingham, AL)

> and application of chemiluminescence ECL substrate

> detection on Hyperfilm ECL autoradiographic film

> (Amersham). For GFAP detection, this method was

> verified in a separate study using a dose response of

> trimethyltin treatment using a GFAP protein standard

> Am. J. Biochem. & Biotech., 4 (2): 218-225, 2008

> 221

> (Chemicon, Inc.). ECL images were scanned into an

> IBM PC using a Hewlett Packard Scanner with a

> transparency adapter. Densiometric analysis was

> performed using Image Pro Analysis Software using

> percent of saline treated controls as the standard.

> Statistical analysis: All behavioral analysis were

> performed using a repeated measures ANOVA

> including both group, day and sex as main factors, with

> the exception of the mid-air righting response which

> was analyzed using Chi-Square and Fisher's Exact Test.

> RESULTS AND DISCUSSION

> Negative geotaxis: Control mice and those treated with

> vitamin E alone were able to perform the reflexive

> negative geotaxis response, reorienting their head to

> point upward when placed on an inclined plane with

> their head facing down. This reflex improved across

> development, as the latency to re-orient improved

> across testing (F (6, 312) = 5.4, p<0.001). VPA-treated

> mice displayed an increased latency to perform this reorientation

> response (F (1, 52) = 10.0, p<0.005). Posthoc

> analysis revealed that following day 14 treatment

> with VPA, there was a significant regression in the

> performance of this response, which reached statistical

> significance on days 16 and 17. Importantly, this VPAinduced

> regression was blocked by vitamin E

> pretreatment, such that pretreated mice were able to

> perform this response with a similar latency as controls

> on P16 and P17 (F (1, 52) = 5.3, p<0.05). Finally, VPAtreated

> mice regained their ability to perform this

> behavioral response similar to controls by the

> completion of testing on P19 (Fig. 1).

> Negative geotaxis

> Day of testing

> 12 13 14 15 16 17 18 19 20

> Latency to re-orient 180 & #61616;C

> 0

> 5

> 10

> 15

> 20

> 25

> 30 Corn oil/saline

> Corn oil/VPA 400

> Vit E/saline

> Vit E/VPA 400

> *

> Fig. 1: Negative geotaxis: Latency to reorient from

> head down to head up on a 45 & #61616;C incline for

> groups of pups treated with VPA (400 mg kg & #61485;1)

> or saline on P14. Some groups received vitamin

> E pretreatment while others received corn oil.

> *: p<0.05 compared to corn oil/saline

> Mid-air righting

> Day of testing

> 12 13 14 15 16 17 18 19 20

> Percent mice able to right in mid-air

> 0

> 20

> 40

> 60

> 80

> 100

> 120 Corn oil/saline

> Corn oil/VPA 400

> Vit E/saline

> Vit E/VPA 400

> *

> Fig. 2: Mid-air righting: Number of pups successfully

> engaging in mid-air righting (expressed as a

> percent of pups mid-air righting on 2 out of 3

> trials/day) for groups of pups treated with VPA

> (400 mg kg & #61485;1) or saline on P14. Some groups

> received vitamin E pretreatment while others

> received corn oil. *: p<0.05 compared to corn

> oil/saline

> Mid-air righting: Before any treatment was

> administered, less than 20% of the pups were able to

> engage in mid-air writing on P13 but this improved to

> about 75% by P14. This observation is interpreted to

> indicate that cerebellar and general muscular maturation

> have matured by P14. & #61539;2 analysis revealed that

> following VPA-treatment given after behavioral testing

> on P14 caused a regression in mid-air righting on P15

> [ & #61539;2 (3) = 39.8, p<0.0001] when compared to saline

> controls. This regression was still observed on P16 in

> the VPA-treated animals. However, the VPA-induced

> regression was eliminated by pretreatment with vitamin

> E (Fig. 2).

> Motor activity: Mice engaged in a similar amount of

> locomotor activity at the start of testing. This activity

> significantly increased across post-natal development

> [F (1, 38) = 80.2, p<0.001]. Interestingly, there was a

> trend for mice treated with VPA to engage in intrusive

> behaviors following P14 treatment. This was evidenced

> by increased levels of activity across testing, beginning

> on P16 through P18. However, this hyperactivity did

> not reach statistical significance. In addition, this

> hyperactive behavior was blocked by pre-treatment

> with vitamin E (Fig. 3).

> GFAP and synaptophysin: Previous work in our lab

> revealed retarded neurobehavioral development in mice

> treated with 600 mg kg & #61485;1 VPA on embryonic day 13[1].

> In order to determine whether we could detect a

> Am. J. Biochem. & Biotech., 4 (2): 218-225, 2008

> 222

> Motor activity:

> Day of testing

> 13 14 15 16 17 18 19 20

> Number of horizontal beam breaks

> 0

> 200

> 400

> 600

> 800

> 1000

> Corn oil+saline

> Corn oil+VPA

> Vitamin E+VPA

> Fig. 3: Motor activity: Horizontal beam breaks for

> groups of pups treated with VPA (400 mg kg & #61485;1)

> or saline on P14. Some groups received vitamin

> E pretreatment while others received corn oil.

> *: p<0.05 compared to corn oil/saline

> Prenatal saline

> Sacrificed day 4

> Prenatal saline

> Sacrificed day 5

> Pprenatal VPA

> Sacrificed day 5

> Glia-fibrillary acidic protein

> 0

> 1000

> 2000

> 3000

> 4000

> 5000

> 6000

> 7000

> SALINE (PND4) SALINE (PND5) VPA (PND5)

> Protein level (arbitrary units)

> *

> Animals treated with VPA 600 mg kg & #61485;1 or Saline on E13

> Fig. 4: GFAP: Pups received either VPA (600 mg kg & #61485;1)

> or saline in utero on E13 and were sacrificed on

> either P4 or P5. *: p<0.05 compared to corn

> oil/saline

> biological marker of VPA-induced neurobehavioral

> retardation, we examined early postnatal levels of

> GFAP and synaptophysin in the cerebellum following

> in utero exposure to VPA. Mice treated with

> 600 mg kg & #61485;1 VPA on E13 and sacrificed on P5 showed

> decreased concentrations of GFAP in the cerebellum

> compared to saline treated controls sacrificed on both

> P4 and P5 (F (2, 8) = 12.3, p<0.01) (Fig. 4). Likewise,

> the concentration of synaptophysin was significantly

> decreased in the cerebellum of E13 VPA-treated mice

> compared to both P4 and P5 saline controls

> Prenatal saline

> Sacrificed day 4

> Prenatal saline

> Sacrificed day 5

> Prenatal VPA

> Sacrificed day 5

> 0

> 1000

> 2000

> 3000

> 4000

> 5000

> 6000

> 7000

> 8000

> SALINE (PND4) SALINE (PND5) VPA (PND5)

> Protein level (arbitrary units)

> *

> Synaptophysin

> Animals treated with VPA 600 mg kg & #61485;1 or Saline on

> E13

> Fig. 5: Synaptophysin: Pups received either VPA

> (600 mg kg & #61485;1) or saline in utero on E13 and

> were sacrificed on either P4 or P5. *: p<0.05

> compared to corn oil/saline

> (F (2, 6) = 10.0, p<0.01) (Fig. 5). During this period of

> postnatal development, GFAP and synaptophysin levels

> increase to promote normal astrocytes-neuron

> interactions and synaptogensis, respectively[37].

> Therefore, mice treated with VPA in utero show

> immature neural development, since the levels of GFAP

> and synaptophysin observed on P5 are much lower than

> those found in mice from an earlier postnatal period

> (P4). This suggests that the behavioral retardations seen

> in our previous study are influenced by retarded neural

> development.

> The etiology of autism is thought to involve early

> exposure to ROS-generating toxicants acting upon

> genetically-sensitive individuals. We have developed a

> new strategy to assess the detrimental effects of early

> toxicant exposure on neurobehavioral development,

> classifying the behavioral deficits as retardations,

> regressions or intrusions. In previous studies we

> demonstrated that early exposure to VPA or MeHg

> results in behavioral deficits in the maturation of social,

> cognitive and motor skills[1,8]. Furthermore, we

> demonstrated that our behavioral model was useful in

> demonstrating that pretreatment with an antioxidant

> protected mice against the behavioral deficits induced

> by early exposure to MeHg[8]. In the present study, we

> demonstrated that vitamin E was capable of protecting

> mice against VPA-induced regression in negative

> geotaxis and mid-air righting as well as against

> intrusive VPA-induced hyperactivity. Collectively, the

> present data together with our previous MeHg study

> indicate that the generation of ROS may be a common

> Am. J. Biochem. & Biotech., 4 (2): 218-225, 2008

> 223

> factor mediating toxicant-induced neuronal damage

> associated with autism and that neurobehavioral

> assessments provide an important functional measure of

> the potential benefits of antioxidants.

> A second objective of the present study was to

> develop a biological marker of the VPA-induced

> damage. Toward this end we used our initial model,

> delivering the VPA prenatally on E13[1]. We had

> demonstrated that this prenatal VPA treatment resulted

> in later behavioral deficits as assessed in the surface

> and mid-air righting tests, negative geotaxis and in

> water maze. Furthermore, this prenatal VPA treatment

> resulted in sex-dependent differences in these

> behavioral deficits with males more affected than

> females. In the present study, we found that both

> cerebellar GFAP and cerebellar synaptophysin were

> reduced postnatally following the prenatal VPA

> administration. GFAP is a marker of astroglia in the

> brain and is involved in astrocyte-neuron interactions.

> GFAP mutant mice have abnormal structure and exhibit

> deficient long-term depression in cerebellar Purkinje

> cell synapses[37]. Therefore, major alterations in GFAP

> may alter Purkinje cell communication that, in turn,

> may alter behavior. Synaptophysin is a widely used

> marker for nerve terminals and can indicate

> synaptogenesis. Therefore, a reduction in

> synaptophysin in the cerebellum could signify a

> reduction in synatpogenesis in that region. More

> generally, it is intriguing that these biological markers

> may reflect the behavioral deficits of cognitive and

> motor retardation caused by the early VPA exposure.

> Future studies are designed to determine if the

> antioxidant pretreatment also protects the mice against

> these neurological changes induced by the VPA.

> There is ample evidence that ROS are involved in

> human autism. Free oxygen radicals could result from

> ingested or inhaled environmental toxins, food or food

> additives, inflammation or infection (overt or occult).

> The interaction of free oxygen radicals and

> polymorphic oxidative genes during gestation or

> postnatally could disrupt neurogenesis in developing

> brain at multiple time windows, eliciting immediate

> stage-dependent effects in specific systems that

> influence subsequent ontogenetic processes, leading to

> the phenotype of autism. Indeed an exacerbated

> oxidative stress response has been implicated in autism.

> Specifically lower glutathione peroxidase (GPX) and

> superoxide dismutase (SOD) activity were found in

> children with autism[38-40]. An increase in body burden

> of various toxins was reported in autism[41,42]. In

> addition, provoked urinary mercury excretion is found

> to be higher in autism[43]. These toxins could generate

> oxidative stress in children with autism. Elevated nitrite

> and nitrate in plasma[44] and red cells[38] have been

> reported in children with autism. This elevation

> indicates excess generation of nitric oxide free radicals.

> In addition, two independent double blind placebo

> controlled clinical trials of antioxidants (vitamin C or

> carnosine) showed beneficial effects in autism[45,46].

> Finally, we conducted a study of oxidative stress

> biomarkers in children with autism and age matched

> healthy controls. Our results showed that urinary

> excretion of 8 isoprostane F2_ was significantly higher

> in children with autism as compared to healthy

> controls[47]. There was also a trend of increased 8-

> OHdG urinary excretion in autistic subjects. These

> results suggest that oxidative stress is exacerbated in

> autism and are consistent with the present results of

> antioxidant protective effects against VPA-induced

> behavioral deficits in mice.

> In summary, we have developed a comprehensive

> neurobehavioral model in which mice are exposed to

> candidate toxicants during critical periods of neural

> development. The mice may have altered expression of

> genes thought to be associated with autism and/or to

> confer increased sensitivity to the toxicants. The mice

> are then assessed in a battery of tests designed to assess

> behavioral maturation of skills in the social, cognitive

> and motor domains. Toxicant or genetic-induced

> deficits in the behavioral maturation are classified as

> retardations, regressions or intrusions. In the present

> studies, we further demonstrate that pretreatment with

> an antioxidant protects the mice against the toxicantinduced

> behavioral deficits. We conclude that our

> model is useful for evaluation of the theory that

> oxidative stress may play a role in the etiology of

> autism.

> ACKNOWLEDGEMENT

> Supported by ES11256, EPAR829391, New Jersey

> Governor's Council on Autism and Autism Speaks.

> REFERENCES

> 1. Wagner, G.C., K.R. Reuhl, M. Cheh, M., P. McRae

> and A.K. Halladay, 2006. A new neurobehavioral

> model of autism in mice: Pre- and Postnatal

> exposure to sodium valproate. J. Autism Dev. Dis.,

> 36: 779-793.

> 2. Bernard, S., A. Enayati, L. Redwood, H. and

> T. Binstock, 2000. Autism: A novel form of

> mercury poisoning. ARC Res., 1-12.

> 3. Dawson, G., 1996. Neuropsychology of autism: a

> report on the state of the science. J. Aut. Dev. Dis.,

> 26: 179.

> Am. J. Biochem. & Biotech., 4 (2): 218-225, 2008

> 224

> 4. Chauhan, A. and V. Chauhan, 2006. Oxidative

> stress in autism. Pathophysiol., 13: 171-181.

> 5. Chauhan, A., V. Chauhan, T. Brown and I. Cohen,

> 2004. Oxidative stress in autism: increased lipid

> peroxidation and reduced serum levels of

> ceruloplasmin and transferin-the antioxidant

> proteins. Life Sci., 75: 2539-2549.

> 6. , S.J., P. Cutler, S. Melnyk, S. Jernigan,

> L. Janak, D.W. Gaylor and J.A. Neubrander, 2004.

> Metabolic biomarkers of increased oxidative stress

> and impaired methylation capacity in children with

> autism. Am. J. Clin. Nutr., 80: 1611-1617.

> 7. Wagner, G.C., K.R. Reuhl, X. Ming and

> A.K. Halladay, 2007. Behavioral and

> neurochemical sensitization to amphetamine

> following early postnatal administration of

> methylmercury. NeuroToxicol., 281: 59-66.

> 8. Cheh, M.A., A.K. Halladay, K.R. Reuhl,

> M. Polunas, X. Ming and G.C. Wagner, 2007.

> Trolox, a Vitamin E derivative, protects against

> persistent neurobehavioral disruption induced by

> neonatal methylmercury exposure. Soc. Toxicol.,

> NC.

> 9. Rodier, P.M., J.L. Ingram, B. Tisdale, S.

> and J. Romano, 1996. Embryological origin for

> autism: developmental abnormalities of the cranial

> nerve motor nuclei. J. Comp. Neurol.,

> 370:247-261.

> 10. Ingram, J.L., S.M. Peckham, B. Tisdale and

> P.M. Rodier, 2000a. Prenatal exposure to valproic

> acid reproduces the cerebellar anomalies associated

> with autism. Neurotox. Teratol., 22, 319-324.

> 11. Sobaniec-Lotoweska, M.E., 2001. Ultrastructure of

> purkinje cell perikara and their dendritic processes

> in the rat cerebellar cortex in experimental

> encephalopathy induced by chronic application of

> valproate. Int. J. Exp. Pathol., 82: 337-348.

> 12. , S.J., P. Turnpenny, A. Quinn, S. Glover

> and D.J. Lloyd, 2000. Montgomery and

> J.C.S. Dean, A clinical study of 57 children with

> fetal anticonvulsant syndromes. J. Med. Genetics,

> 37: 489-497.

> 13. , G., J. King, M. Cunningham,

> M. Stephan, B. Kerr and J.H. Hersh, 2001. Fetal

> valproate syndrome and autism: additional

> evidence of an association. Dev. Med. Child

> Neurol., 43: 202-206.

> 14. Chapman, J.B. and M.G. Cutler, 1989. Effects of

> sodium valproate on development and social

> behaviour in the Mongolian gerbil. Neurotoxicol.

> Teratol., 11: 193-198.

> 15. Voorhees, C.V., 1987. Behavioral teratogenicity of

> valproic acid: selective effects on behavior after

> prenatal exposure to rats. Psychopharmacol.,

> 92: 173-179.

> 16. Schneider, T. and R. Przewlocki, 2005. Behavioral

> alterations in rats prenatally exposed to valproci

> acid: Animal model of autism.

> Neuropsychopharmacol., 30: 80-89.

> 17. Altman, J. and S.A. Bayer, 1978. Prenatal

> development of the cerebellar system in the rat.

> Cytogenesis and histogenesis of the deep nuclei

> and cortex of the cerebellum. J. Comp. Neurol.,

> 179: 23-48.

> 18. Inouye, M. and U. Murakami, 1980. Temporal and

> spatial patterns of purkinje cell formation in the

> mouse cerebellum. J. Comp. Neurol.,

> 194: 499-503.

> 19. Bachevalier, J. and M. Beauregard, 1993.

> Maturation of medial temporal lobe memory

> functions in rodents, monkeys and humans.

> Hippocampus, 3:191-202.

> 20. Rice, D. and S. Barone, 2000. Critical periods of

> vulnerability for the developing nervous system:

> Evidence from human and animal models. Environ.

> Health Per., 108: 511-533.

> 21. Murugesan, V. and P. Subramanian, 2003.

> Enhancement of circulatory antioxidants by & #61537;ketoglutarate

> during sodium valproate treatment in

> wistar rats. Polish J. Pharmacol., 55: 31-36.

> 22. Tong, V., T.K.H. Chang, J. Chen and F.S. Abbott,

> 2003. The effect of valproic acid on hepatic and

> plasma levels of 15-F2t-Isoprostane in rats. Free

> Radical Biol. Med. 34: 1435-1446.

> 23. Sobaniec-Lotowska, M.E., 1997. Effects of longterm

> administration of the antiepileptic drugsodium

> valproate upon the ultrastructure of

> hepatocytes in rats. Exp. Toxicol. Pathol.,

> 49: 225-32.

> 24. Myers, G.J. and P.W. son, 2000. Does

> methylmercury have a role in causing

> developmental disabilities in children? Environ.

> Health Per., 108: 413-420.

> 25. Rice, D.C., 1996. Sensory and cognitive effects of

> developmental methylmercury exposure in

> monkeys and a comparison to effects in rodents.

> Neurotoxicol., 17, 139-154.

> 26. Dey, P.M. and K.R. Reuhl, 1999. Developmental

> methylmercury administration alters cerebellar

> PSA-NCAM expression and Golgi sialyltransferase

> activity. Res., 845:139-151.

> 27. Yee, S. and B.H. Choi, 1996. Oxidative stress in

> neurotoxic effects of methylmercury poisoning.

> Neurotoxicol., 17: 17.

> Am. J. Biochem. & Biotech., 4 (2): 218-225, 2008

> 225

> 28. Park, S.T., K.T. Lim, Y.T. Chung and S.U. Kim,

> 1996. Methylmercury-induced neurotoxicity in

> cerebral neuron culture is blocked by antioxidants

> and NMDA receptor antagonists. Neurotoxicol.,

> 17: 37.

> 29. Usuki, F., A. Yasutake, F. Umehara, H. Tokunaga

> and M. Matsumoto et al., 2001. In vivo protection

> of a water-soluble derivative of vitamin E, Trolox,

> against methylmercury-intoxication in the rat

> Neurosci Lett., 304: 199-203.

> 30. Sanfeliu, C., J. Sebastia and S.U. Ki, 2001.

> Methylmercury neurotoxicity in cultures of human

> neurons, astrocytes, neuroblastoma cells.

> Neurotoxicol., 22: 317-327.

> 31. Aschner, M., C.P. Yao, J.W. and K.H. Tan,

> 2000. Methylmercury alters glutamate transport in

> astrocytes. Neurochem. Int., 37: 19.

> 32. Stohs, S.J. and D. Bagchi, 1995. Oxidative

> mechanisms in the toxicity of metal ions. Free

> Radical Biol. Med., 18: 321.

> 33. Sorg, O., B. Schilter, P. Honegger and

> F. Monnet-Tschudi, 1998. Increased vulnerability

> of neurones and glial cells to low concentrations of

> methylmercury in a prooxidant situation. Acta

> Neuropathol. (Berl), 96: 621.

> 34. Mundy, W.R. and T.M. Freudenrich, 2000.

> Sensitivity of immature neurons in culture to

> metal-induced changes in reactive oxygen species

> and intracellular free calcium. Neurotoxicol,

> 21: 1135.

> 35. Altman, J. and K. Sudarshan, 1975. Postnatal

> development of locomotion in the laboratory rat.

> Animal Behav., 23: 896-901.

> 36. Petrosini, L., M. Molinari and T. Gremoli, 1990.

> Hermicerebellectomy and motor behavior in rats.

> II. Effects of cerebellar lesion performed at

> different developmental stages. Exp. Brain Res.,

> 82: 483-492.

> 37. Shibuki, K., H. Gomi, L. Chen, S. Bao, J.J. Kim,

> H. Wakatsuki, T. Fujisaki, K. Fujimoto, A. Katoh,

> T. Ikeda, C. Chen, R.F. and S. Itohara,

> 1996. Deficient cerebellar Long-term depression,

> impaired eyeblink conditioning and normal motor

> coordination in GFAP mutant mice. Neuron,

> 16: 587-599.

> 38. Sogut, S., S.S. Zoroglu and H. Ozyurt et al., 2003.

> Changes in nitric oxide levels and antioxidant

> enzyme activities may have a role in the

> pathophysiological mechanisms involved in

> autism. Clinica Chimica Acta, 331: 111-117.

> 39. Golse, B., P. Debray-Ritzen and P. Durosay et al.,

> 1978. Perturbation de deux enzymes, la

> superoxyde-dismutase I et la glutathioneperoxydase

> dans la psychose infantile de

> developpement (autisme infantile). Rev. Neurol.

> (Paris), 134: 699-705.

> 40. Yorbik, O., A. Sayal and C. Akay et al., 2002.

> Investigation of antioxidant enzymes in children

> with autistic disorder. Prostaglandins Leukot

> Essent Fatty Acids, 67: 341-343.

> 41. Edelson, S.B. and D.S. Cantor, 1998. Autism:

> xenobiotic influences. Toxicol. Ind. Health, 14:

> 799-811.

> 42. Edelson S.B. and D.S. Cantor, 2000. The

> neurotoxic etiology of the autistic spectrum

> disorder: a replicative study. Toxicol. Ind. Health,

> 16: 239-47.

> 43. Bradstreet, J., D.A. Geier and J.J. Kartzinel et al.,

> 2003. A case-control study of mercury burden in

> children with autistic spectrum disorders. J. Am.

> Phy. Sur., 8: 76-79.

> 44. Zoroglu, S.S., M. Yurekli and I. Meram et al.,

> 2003. Pathophysiological role of nitric oxide and

> adrenomedullin in autism. Cell Biochem. Function,

> 21: 55-60.

> 45. Chez, M.D., C.P. Buchanan and M.C. Aimonovitch

> et al., 2002. Double-blind, placebo-controlled

> study of l-carnosine supplementation in children

> with autistic spectrum disorders. J. Child Neurol.,

> 17: 833-837.

> 46. Dolske, M.C., J. Spollen and S. McKay et al.,

> 1993. A preliminary trial of ascorbic acid as

> supplemental therapy for autism. Prog. Neuro-

> Psychopharm. Biol. Psychiat., 17: 765-774.

> 47. Ming, X., P. Stein, M. Brimacombe, W. ,

> G. Lambert and G.C. Wagner, 2005. Increased

> excretion of a lipid peroxidation biomarker in

> autism. Prostaglandins, Leukotrienes and Essential

> Fatty Acids, 73: 379-384.

> ~~~~~~~~~~~

>

> =====

>

February 23, 2008

Certainly relevant to this discussion.... i'm ALL FOR giving weight

to all sides before jumping on any band wagon.

I HIGHLY suggest everyone reading this line of posts take a peak at

this article. Rather, read the whole thing. Very interesting

http://www.ageofautism.com/2008/02/the-aap-knows-v.html

In fact, www.ageofautism - in my book simply tells it like it is,

everyday there is something new to ponder.

Happy reading.

> > From the study I just posted:

> > " Nonetheless, it is important to note that autism was not found

to be

> > associated with either pre- or neonatal exposure to

> > organic mercury. "

> Included in the cited report or not - this is NOT a true

statement....no

> 100% conclusive causal link has been found NOR HAS IT BEEN

DISPROVED at

> this point by anybody that organic mercury (Thimerosal) ALONE

triggers

> or doesn't trigger autism/autoimmune problems...but studies are

still in

> progress for both sides of the discussion...and the " causes " are

not

> focused on a single environmental contaminant (i.e. organic

> mercury/thimerosal)...there is agreement that it is a NEUROtoxin

and

> what damage a NEUROtoxin does to the head of the NEUROlogical

system is

> pretty obvious...it kills NEUROns and does create a damage to the

> NEUROlogical system. It is only one of many toxins in pre- or

neonatal

> exposures that contributes to lots of health issues.

>

> What gets said publicly and by who depends on who's reporting it

and who

> is getting paid by or receiving research grants or advertising

monies

> from the very pharmaceutical companies who continue to produce

vaccines

> with thimerosal (organic mercury)...leaving even traces (while

> officially not using it labeled as a " preservative " but it is still

used

> during the production phase and " filtered " out leaving levels which

are

> still 1000 times more than the EPAs minimum allowed ppm (parts per

> million) for drinking water. (anyone new who wants to know more

can

> check out the group EOHarm but warning that it is much more

high

> volume than this apraxia group). It's just ONE facet of how

> environmental toxins in general are messing with humans.

>

> If the door is opened to discuss this subject - be prepared before

> presenting it as a " fact " especially if you have not yet read up

on ALL

> the research on both sides currently out there. I'm sure other

biomed

> moms will reply publically to this...so that all the other members

don't

> get misled. Those of us who've been researching all of this

stuff

> for our own kids for years will beg to differ based on validity of

the

> statement considering the 100's of scientific research reports

> (including published ones in scientific journals) and ongoing

> investigative reports we've made time to locate and read in their

> entirety...oh and the documented medical problems, including

apraxia,

> that our kids are forced to overcome.

>

> What you pointed out, truth or mistruth, is a subject much larger

and

> controversial than you may have possibly imagined and

> is probably not the best venue to continue it,

but

> will probably become yet another " hot topic " while things get

clarified

> yet again for the new families coming into the discussion. " Hot

> topics " seem to be the trend this weekend already though.

>

> ____________________________________________________________

> Receive Notifications of Incoming Messages

> Easily monitor multiple email accounts & access them with a click.

> Visit http://www.inbox.com/notifier and check it out!

>

February 23, 2008

Any study in this area helps because studies take funding. We need

to get funding for validation to prove what we already know so that

others can't continue to misinform the public. Imagine years ago

with people that had scurvy that some felt they needed to give a host

of supplements when all they needed was the vitamin C.

Not sure why some of you area jumping on the fact that it may

not 'just' be mercury that's an issue. I'm very aware that I pulled

it out of context which was in direct response to the (?) message

from Liz that mercury causes autism so what causes apraxia. Anyone

who doesn't know that apraxic children were also looked at but not

mentioned is not in reality. Carolyn below in the archives for

example was one of Dr. Ming's patients and I know that they did check

at that time cheek cell scrapings from all -but in the end the

funding reasons typically 'only' mentions autism which is very sad for the

even larger number of apraxic children. Also... I left the entire

published paper intact right underneath where one can learn that

there are other toxins to worry about. In this group the toxin issue

has been up for conversation for years

http://www.cherab.org/news/Save.html -and Dr. Xue Ming from this

study was one of the MDs I was asked to present to at UMDNJ back in

2002 on my theory of how toxins cross the placenta while we are

pregnant. Here's just one post about it from then:

Re: brain & efa's!

Caroline that's amazing!!! I know you were planning to -did you see

Dr. Ming yet at UMDNJ and what did she have to say about this? I

just posted about her/this re: MS and said how Dr. Ming was studying

the

remyelination for MS patients. I know this is one of the areas in

autistic/apraxic children that she/UMDNJ was interested in exploring

more...looking into the remylenating properties of the EFAs. I'm

curious to see

if there is some type of environmentally based myelin damage -

even if it's subtle - that is creating the dramatic rise in children

presenting with communication impairments. I know UMDNJ was even

looking at those children who's myelin problem was not severe enough

to even show up on an MRI. However with a child like yours who has

documented myelin challenges -this is such incredible news -you must

be thrilled! How is your daughter doing? I know improvements were

coming -do you still have surges and where is she now?

I miss talking to you since I've moved to Florida -I'll try to give

you a call -or you can still reach me on my NJ cell phone number if

you have that.

=====

February 23, 2008

So pure apraxia, what you have seen mostly over the years, starts in

utero typically? I ask this because during our long and winding

journey over the past three years one of the many things I saw in my

son that never matched up anywhere, was sort of this " failure to get

out of the gate " completely, from day one but his only true delays

were speech and tone. He is a puzzle for sure and thankfully a

healing one at that but I guess what I am asking is, when you talk

about apraxia, true apraxia apart from any other condition, comorbid

or not, is the deal that it was there from the start as opposed to

other things that are seen in other dx, like regressive autism? late

forming CP? etc. or traumatic brain injuries causing speech and other

delays.

>

> Any study in this area helps because studies take funding. We need

> to get funding for validation to prove what we already know so that

> others can't continue to misinform the public. Imagine years ago

> with people that had scurvy that some felt they needed to give a

host

> of supplements when all they needed was the vitamin C.

>

> Not sure why some of you area jumping on the fact that it may

> not 'just' be mercury that's an issue. I'm very aware that I pulled

> it out of context which was in direct response to the (?) message

> from Liz that mercury causes autism so what causes apraxia. Anyone

> who doesn't know that apraxic children were also looked at but not

> mentioned is not in reality. Carolyn below in the archives for

> example was one of Dr. Ming's patients and I know that they did

check

> at that time cheek cell scrapings from all -but in the end the

> funding reasons typically 'only' mentions autism which is very sad

for the

> even larger number of apraxic children. Also... I left the entire

> published paper intact right underneath where one can learn that

> there are other toxins to worry about. In this group the toxin

issue

> has been up for conversation for years

> http://www.cherab.org/news/Save.html -and Dr. Xue Ming from this

> study was one of the MDs I was asked to present to at UMDNJ back in

> 2002 on my theory of how toxins cross the placenta while we are

> pregnant. Here's just one post about it from then:

>

> Re: brain & efa's!

>

> Caroline that's amazing!!! I know you were planning to -did you see

> Dr. Ming yet at UMDNJ and what did she have to say about this? I

> just posted about her/this re: MS and said how Dr. Ming was studying

> the

> remyelination for MS patients. I know this is one of the areas in

> autistic/apraxic children that she/UMDNJ was interested in exploring

> more...looking into the remylenating properties of the EFAs. I'm

> curious to see

> if there is some type of environmentally based myelin damage -

> even if it's subtle - that is creating the dramatic rise in children

> presenting with communication impairments. I know UMDNJ was even

> looking at those children who's myelin problem was not severe enough

> to even show up on an MRI. However with a child like yours who has

> documented myelin challenges -this is such incredible news -you must

> be thrilled! How is your daughter doing? I know improvements were

> coming -do you still have surges and where is she now?

>

> I miss talking to you since I've moved to Florida -I'll try to give

> you a call -or you can still reach me on my NJ cell phone number if

> you have that.

>

> =====

>

February 23, 2008

<<Not sure why some of you area jumping on the fact that it may

not 'just' be mercury that's an issue. I'm very aware that I pulled

it out of context which was in direct response to the (?) message

from Liz that mercury causes autism so what causes apraxia.>>

This must be a product of all the quick reading and posting today,

but I neither know to what you are referring in your first statement

above ( " jumping on the fact that it may not 'just " be mercury... " )

nor did I understand that to be what Liz was saying or asking. :-)

But, anyway...

This new study does appear to be leading to more new autism studies -

- it actually suggests as much as it holds up its animal model as an

effective way to replicate " autism " in animals for research

purposes.

>

> Any study in this area helps because studies take funding. We need

> to get funding for validation to prove what we already know so that

> others can't continue to misinform the public. Imagine years ago

> with people that had scurvy that some felt they needed to give a

host

> of supplements when all they needed was the vitamin C.

>

> Not sure why some of you area jumping on the fact that it may

> not 'just' be mercury that's an issue. I'm very aware that I

pulled

> it out of context which was in direct response to the (?) message

> from Liz that mercury causes autism so what causes apraxia. Anyone

> who doesn't know that apraxic children were also looked at but not

> mentioned is not in reality. Carolyn below in the archives for

> example was one of Dr. Ming's patients and I know that they did

check

> at that time cheek cell scrapings from all -but in the end the

> funding reasons typically 'only' mentions autism which is very sad

for the

> even larger number of apraxic children. Also... I left the entire

> published paper intact right underneath where one can learn that

> there are other toxins to worry about. In this group the toxin

issue

> has been up for conversation for years

> http://www.cherab.org/news/Save.html -and Dr. Xue Ming from this

> study was one of the MDs I was asked to present to at UMDNJ back in

> 2002 on my theory of how toxins cross the placenta while we are

> pregnant. Here's just one post about it from then:

>

> Re: brain & efa's!

>

> Caroline that's amazing!!! I know you were planning to -did you see

> Dr. Ming yet at UMDNJ and what did she have to say about this? I

> just posted about her/this re: MS and said how Dr. Ming was

studying

> the

> remyelination for MS patients. I know this is one of the areas in

> autistic/apraxic children that she/UMDNJ was interested in

exploring

> more...looking into the remylenating properties of the EFAs. I'm

> curious to see

> if there is some type of environmentally based myelin damage -

> even if it's subtle - that is creating the dramatic rise in

children

> presenting with communication impairments. I know UMDNJ was even

> looking at those children who's myelin problem was not severe

enough

> to even show up on an MRI. However with a child like yours who has

> documented myelin challenges -this is such incredible news -you

must

> be thrilled! How is your daughter doing? I know improvements were

> coming -do you still have surges and where is she now?

>

> I miss talking to you since I've moved to Florida -I'll try to give

> you a call -or you can still reach me on my NJ cell phone number if

> you have that.

>

> =====

>

February 23, 2008

I just checked in to see if I was missing anything, so I'm not up to

speed on this thread.... but I wanted to mention that I had my hair

and my daughter's hair tested, expecting mercury because my mom had

high levels of mercury (I figured she could have passed it to me).

Anyway, turns out my Uranium (from Radon probably) was off the charts,

and my daughters was 1 1/2 times mine. So you might be right, and it

might not be mercury, but that doesn't mean it's not something else!

It seems like different things are the triggers in different cases,

but if you look it seems like it's usually some sort of toxin built

up. Just my .02

Crystal

>

> I'm not sure whether my thoughts mean much or anything-- but I've

never been

> exposed to mercury during pregnancies-- we don't vaccinate, and I

personally

> was never exposed to any mercury, so for MY son's pregnancy-- this

theory of

> mercury causing Apraxia-- wouldn't be valid.

>

> Becky

>

February 24, 2008

RE: Uranium

There is a little girl on the board, globally apraxic, who came up

high in uranium, as did her house water.

RE: Mercury

I have never heard of mercury causing apraxia and am not sure that is

what was said here. As for mercury exposure...depends what you mean.

If you have mercury amalgams, went to the dentist, live in NJ (LOL)

there was likely some exposure. It is in the air. I live down the

street from a pharmaceutical plant that manufactures vaccines so it

is here and to be truthful I do not even think mercury is our big

concern.

Whatever this is that has hit our kids is multifaceted, nature,

nurture, and the rest and each is a puzzle...likely the reason we

fight over remedies.

>

> I just checked in to see if I was missing anything, so I'm not up to

> speed on this thread.... but I wanted to mention that I had my hair

> and my daughter's hair tested, expecting mercury because my mom had

> high levels of mercury (I figured she could have passed it to me).

> Anyway, turns out my Uranium (from Radon probably) was off the

charts,

> and my daughters was 1 1/2 times mine. So you might be right, and

it

> might not be mercury, but that doesn't mean it's not something

else!

> It seems like different things are the triggers in different cases,

> but if you look it seems like it's usually some sort of toxin built

> up. Just my .02

>

> Crystal

>

> >

> > I'm not sure whether my thoughts mean much or anything-- but I've

> never been

> > exposed to mercury during pregnancies-- we don't vaccinate, and I

> personally

> > was never exposed to any mercury, so for MY son's pregnancy--

this

> theory of

> > mercury causing Apraxia-- wouldn't be valid.

> >

> > Becky

> >

>

February 24, 2008

Please do not misunderstand me. I have many theories behind what we

have seen in my family and I assure you metal is only one part and

may likely be a small but important part is all.

>

> In a message dated 2/23/2008 7:04:03 P.M. Eastern Standard Time,

> crystalam_@... writes:

>

> (I figured she could have passed it to me).

> Anyway, turns out my Uranium (from Radon probably) was off the

charts,

> and my daughters was 1 1/2 times mine. So you might be right, and

it

> might not be mercury, but that doesn't mean it's not something

else!

> It seems like different things are the triggers in different cases,

> but if you look it seems like it's usually some sort of toxin built

> up. Just my .02

>

> Again-- In my case-- my toxin levels were fine. No uranium

problems etc.

> I am TOTALLY hearing what you're saying though, and AGREE that many

times

> the cause of these neurological based disabilities are exposures to

metals and

> toxins. (but will say definitely NOT ALL)

>

> becky

>

> **************Ideas to please picky eaters. Watch video on AOL

Living.

> (http://living.aol.com/video/how-to-please-your-picky-eater/rachel-

campos-duffy/

> 2050827?NCID=aolcmp00300000002598)

>

February 24, 2008

I just meant generally. Apparently they tested the air where I

live...high in mercury and lead!

>

> In a message dated 2/23/2008 7:18:58 P.M. Eastern Standard Time,

> lizlaw@... writes:

>

> As for mercury exposure...depends what you mean.

> If you have mercury amalgams, went to the dentist, live in NJ

(LOL)

> there was likely some exposure.

>

> You're probably thinking with THIS current pregnancy-- and not

Asa's. ????

> With THIS pregnancy-- I've been to the dentist and the mercury

fillings on

> one side taken out, so the concern would be present NOW for THIS

pregnancy.

> With ASA though, who is my Apraxic child--- there definitely

wasn't any of

> these issues.

>

> Becky

>

> **************Ideas to please picky eaters. Watch video on AOL

Living.

> (http://living.aol.com/video/how-to-please-your-picky-eater/rachel-

campos-duffy/

> 2050827?NCID=aolcmp00300000002598)

>

February 24, 2008

Well,

my son is a Lead kid and has problems with mercury too.....

Janice

[sPAM]Re: [ ] Re: New study that validates vitamin

E!!!!!

In a message dated 2/23/2008 7:04:03 P.M. Eastern Standard Time,

crystalam_@... writes:

(I figured she could have passed it to me).

Anyway, turns out my Uranium (from Radon probably) was off the charts,

and my daughters was 1 1/2 times mine. So you might be right, and it

might not be mercury, but that doesn't mean it's not something else!

It seems like different things are the triggers in different cases,

but if you look it seems like it's usually some sort of toxin built

up. Just my .02

Again-- In my case-- my toxin levels were fine. No uranium problems etc.

I am TOTALLY hearing what you're saying though, and AGREE that many times

the cause of these neurological based disabilities are exposures to metals and

toxins. (but will say definitely NOT ALL)

becky

**************Ideas to please picky eaters. Watch video on AOL Living.

(http://living.aol.com/video/how-to-please-your-picky-eater/rachel-campos-duffy/

2050827?NCID=aolcmp00300000002598)

February 24, 2008

<<Anyone who doesn't know that apraxic children were also looked at

but not mentioned is not in reality.>>

What are you trying to say?

I came back to re-read this study and discussion this a.m., because

right now, I'm reviewing and reconsidering a number of things, most

notably Vitamin E (and The Listening Program and NACD.) (Our initial

experience with Vit. E was bad. But, we're still searching and may

re-try in the near term.)

In doing so, I saw this statement and, again, became confused.

Because your comment above was posted in response to my questions

about the applicability of the study you provided as a " New study

that validates vitamin E, " I am following up and would like to

understand what you mean.

If you have a moment, will you please explain? Thank you in advance.

>

> Any study in this area helps because studies take funding. We need

> to get funding for validation to prove what we already know so that

> others can't continue to misinform the public. Imagine years ago

> with people that had scurvy that some felt they needed to give a

host

> of supplements when all they needed was the vitamin C.

>

> Not sure why some of you area jumping on the fact that it may

> not 'just' be mercury that's an issue. I'm very aware that I

pulled

> it out of context which was in direct response to the (?) message

> from Liz that mercury causes autism so what causes apraxia. Anyone

> who doesn't know that apraxic children were also looked at but not

> mentioned is not in reality. Carolyn below in the archives for

> example was one of Dr. Ming's patients and I know that they did

check

> at that time cheek cell scrapings from all -but in the end the

> funding reasons typically 'only' mentions autism which is very sad

for the

> even larger number of apraxic children. Also... I left the entire

> published paper intact right underneath where one can learn that

> there are other toxins to worry about. In this group the toxin

issue

> has been up for conversation for years

> http://www.cherab.org/news/Save.html -and Dr. Xue Ming from this

> study was one of the MDs I was asked to present to at UMDNJ back in

> 2002 on my theory of how toxins cross the placenta while we are

> pregnant. Here's just one post about it from then:

>

> Re: brain & efa's!

>

> Caroline that's amazing!!! I know you were planning to -did you see

> Dr. Ming yet at UMDNJ and what did she have to say about this? I

> just posted about her/this re: MS and said how Dr. Ming was

studying

> the

> remyelination for MS patients. I know this is one of the areas in

> autistic/apraxic children that she/UMDNJ was interested in

exploring

> more...looking into the remylenating properties of the EFAs. I'm

> curious to see

> if there is some type of environmentally based myelin damage -

> even if it's subtle - that is creating the dramatic rise in

children

> presenting with communication impairments. I know UMDNJ was even

> looking at those children who's myelin problem was not severe

enough

> to even show up on an MRI. However with a child like yours who has

> documented myelin challenges -this is such incredible news -you

must

> be thrilled! How is your daughter doing? I know improvements were

> coming -do you still have surges and where is she now?

>

> I miss talking to you since I've moved to Florida -I'll try to give

> you a call -or you can still reach me on my NJ cell phone number if

> you have that.

>

> =====

>

February 24, 2008

Hey - Any new studies like in the last 2 years of your claims...because

there are a lot of parents here who can attest...just as you have on many

issues, and have brought new studies to the table, that you continue to " POO

POO " ???? Back to the " MAJORITY " conversation...who is the majority? Because

since Ive been on the list....all the new parents ARE now the

MAJORITY....your MAJORITY from back in 2001, 2002, and 2003...where are

they? We all get that your children are NOT AUTISTIC, you do not have to

continue to bang your head against the wall any longer proving it !!!

Thanks

Michele

[ ] Re: New study that validates vitamin E!!!!!

<<Anyone who doesn't know that apraxic children were also looked at

but not mentioned is not in reality.>>

What are you trying to say?

I came back to re-read this study and discussion this a.m., because

right now, I'm reviewing and reconsidering a number of things, most

notably Vitamin E (and The Listening Program and NACD.) (Our initial

experience with Vit. E was bad. But, we're still searching and may

re-try in the near term.)

In doing so, I saw this statement and, again, became confused.

Because your comment above was posted in response to my questions

about the applicability of the study you provided as a " New study

that validates vitamin E, " I am following up and would like to

understand what you mean.

If you have a moment, will you please explain? Thank you in advance.

>

> Any study in this area helps because studies take funding. We need

> to get funding for validation to prove what we already know so that

> others can't continue to misinform the public. Imagine years ago

> with people that had scurvy that some felt they needed to give a

host

> of supplements when all they needed was the vitamin C.

>

> Not sure why some of you area jumping on the fact that it may

> not 'just' be mercury that's an issue. I'm very aware that I

pulled

> it out of context which was in direct response to the (?) message

> from Liz that mercury causes autism so what causes apraxia. Anyone

> who doesn't know that apraxic children were also looked at but not

> mentioned is not in reality. Carolyn below in the archives for

> example was one of Dr. Ming's patients and I know that they did

check

> at that time cheek cell scrapings from all -but in the end the

> funding reasons typically 'only' mentions autism which is very sad

for the

> even larger number of apraxic children. Also... I left the entire

> published paper intact right underneath where one can learn that

> there are other toxins to worry about. In this group the toxin

issue

> has been up for conversation for years

> http://www.cherab.org/news/Save.html -and Dr. Xue Ming from this

> study was one of the MDs I was asked to present to at UMDNJ back in

> 2002 on my theory of how toxins cross the placenta while we are

> pregnant. Here's just one post about it from then:

>

> Re: brain & efa's!

>

> Caroline that's amazing!!! I know you were planning to -did you see

> Dr. Ming yet at UMDNJ and what did she have to say about this? I

> just posted about her/this re: MS and said how Dr. Ming was

studying

> the

> remyelination for MS patients. I know this is one of the areas in

> autistic/apraxic children that she/UMDNJ was interested in

exploring

> more...looking into the remylenating properties of the EFAs. I'm

> curious to see

> if there is some type of environmentally based myelin damage -

> even if it's subtle - that is creating the dramatic rise in

children

> presenting with communication impairments. I know UMDNJ was even

> looking at those children who's myelin problem was not severe

enough

> to even show up on an MRI. However with a child like yours who has

> documented myelin challenges -this is such incredible news -you

must

> be thrilled! How is your daughter doing? I know improvements were

> coming -do you still have surges and where is she now?

>

> I miss talking to you since I've moved to Florida -I'll try to give

> you a call -or you can still reach me on my NJ cell phone number if

> you have that.

>

> =====

>

February 24, 2008

crystal - this is a very common problem with the hair test. If you

DONT see mercury on the report - and your kid is symtomatic of course-

you can pretty much assume you kid's body is holding on to it- thus,

still could be an issue. So, this is a good reason to have a

seasoned professional interpret the hair analysis. It's really not

your " best " gage anyway- it can tell you if your kid is holding - and

the metals that are coming out - evidenced by the metals that show in

high ranges. The french urin porphyrin test is a much better gage

of mercury and lead load - it also shows pcbs, pesticides. I have 2

hair analysis - which even with a seasoned pro interpretation - I

found the French urin porphryine to be the most telling.

>

> RE: Uranium

> There is a little girl on the board, globally apraxic, who came up

> high in uranium, as did her house water.

Sign In

Re: New study that validates vitamin E!!!!!

Recommended Posts

Guest guest

Link to comment

Share on other sites

Guest guest

Link to comment

Share on other sites

Guest guest

Link to comment

Share on other sites

Guest guest

Link to comment

Share on other sites

Guest guest

Link to comment

Share on other sites

Guest guest

Link to comment

Share on other sites

Guest guest

Link to comment

Share on other sites

Guest guest

Link to comment

Share on other sites

Guest guest

Link to comment

Share on other sites

Guest guest

Link to comment

Share on other sites

Guest guest

Link to comment

Share on other sites

Guest guest

Link to comment

Share on other sites

Guest guest

Link to comment

Share on other sites

Guest guest

Link to comment

Share on other sites

Guest guest

Link to comment

Share on other sites

Guest guest

Link to comment

Share on other sites

Guest guest

Link to comment

Share on other sites

Guest guest

Link to comment

Share on other sites

Guest guest

Link to comment

Share on other sites

Guest guest

Link to comment

Share on other sites

Guest guest

Link to comment

Share on other sites

Guest guest

Link to comment

Share on other sites

Guest guest

Link to comment

Share on other sites

Guest guest

Link to comment

Share on other sites

Guest guest

Link to comment

Share on other sites

Join the conversation

Activity