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Note by D. Grant, Ph.D., (Research Chemist) Turriff, Scotland, United
Kingdom (U.K.) AB53 6SX 3/11/09-14/2/10 a
This document is not intended as a neat finished scientific discourse, it is more of a preliminary rough listing of ideas
gleaned from reading the literature on the internet. Some of these may be of interest to others.
c

.
This article is not intended as a substitute for medical advice

it is however intended to stimulate debate and identify possible ways of combating illness in the future

., it is for similar purpose as the author¶s hypothesis building articles which have previously appeared in peer-
reviewed scientific journals.
The need for open discussions of this kind in regard to MNDs especially
arises 
. from the statement below
³V

 
V

V

V
V ‰.
which appears on the Motor Neuron Information Page of the US National Institute of
Neurological Disorders and Stroke (NINDS).

[While the U.S. Food & Drug Administration has currently approved Riluzole for the
treatment ALS*/MND,
it should be noted this drug has been found to offer only very modest benefit].

Considerations of literature available on possible tissue protective functions

of heparin/heparan sulfate leads to the questions:
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The scientific literature accessed from the perspective of polysaccharide
biochemistry could suggest that sporadic ALS*, as well as other neurological
degenerative disease processes occur, at least in part, as a consequence of
deficiencies in linked antioxidant/nitrant defense, where protein tyrosine
nitration occurs and this arises, at least in part, as a consequence of
deficiencies in heparan sulfate (HS) ±related wide range tissue protection
system status.
A recent report (M.L.T. Teoh c., Cancer Res., 2009, 69, 6355-6363)
suggested the existence of a linkage between redox status (oxidant and nitrant
overload) and HS activity.
These authors found that overexpression of the HS-binding SOD attenuated heparanase
expression (which further inhibited breast cancer cell growth and invasion) this could also
indicate that oxidative stress (
., following superoxide dismutase (SOD) deficiency)
would likely augment heparanase expression.
c

ALS, however, generally is believed to arise from glutamate excitotoxicity

arsing from a general nitric oxide and antioxidant deficiency dependent
selective motor neuron death.

The historical breakthrough in the understanding that ALS was the discovery
that Cu,Zn SOD (intracellular SOD-1) underwent mutations in familial ALS
(fALS).
The (SOD deficiency) mouse model of fALS is, however associated with the
occurrence of widespread protein nitration (F. Casoni, J. Biol. Chem., 2005
280 (16) 16295-16304).
A (seven fold) increase in the presence of 3-nitrotyrosine had been earlier
reported (H. Tohgi c., Ann. Neurol., 1999, 46, 129-131) to occur in the
cerebrospinal fluid of sporadic ALS patients relative to that of healthy
controls.
Pathological nitration following nitrant overload in ALS can, it should also be noted,
explain why tamoxifen (which forms potent antioxidant metabolites) is apparently able to
prolong the lives of ALS victims who are also receiving tamoxifen therapy for breast
cancer (internet report 
c ) and probably a similar mechanism enables estradiol
(which can mop up nitrite) to protects cultured spinal neurons from glutamate and nitric
oxide induced selective motor neuronal death (this was reported by T. Nakamizo c.,
Neuro Report, 2000, 11 (16) 3493-3494).

The abundant serum proteins which may be nitrated under oxidative stress include alpha-
2-macroglobulin (a2M) (which with apo-lipoprotein A-1) seem to serve as scavengers of
reactive oxygen and nitrogen.
(
B. Ghesquiere c., 2009 MCP Ms M900259-MCP200).

The anticoagulant and putatively also the cytokine regulatory roles (which includes
heparin/HS regulation of thrombin and a2M of transforming growth factor beta-1 (TGFi-
1)) of a2M might be compromised by excessive nitration.
The control systems for nitric oxide formation, including the signaling for augmented inducible nitric
oxide synthase (iNOS) activity and its modulation by 
. by actions of transforming growth factor beta
(TGF-i) may therefore be defective in ALS. [The findings of J. I. Zecka c. (Cytokine, 2002, 20 (5)
239-243) suggest that in ALS an altered TGFi-1 activity is associated with the neurodegeneration
process, being higher in patients with a longer duration of ALS].
ALS can apparently be distinguished from other neurodegenerative disease processes from cytokine
biomarker profiling;
ALS seems to be most characteristically associated with an elevation of IL-17 (›
, R.M. Mitchell c.,
Neurology, 2009, 72, 14-19); also by IL-8 (which is induced by nitric oxide).

While the kind of SOD mutations originally identified as occurring in fALS are now
apparently generally thought to create proteins which are more prone to misfolding and
resultant pro-oxidant fibril formation, there is still confusion about the possible key role of
diminished antioxidant protection in ALS; it remains highly likely that this is a principal
defect leading to neurodegeneration (›. H. Ischiropoulos and J.S. Beckman, J. Clin.
Invest., 2003, 111 (2) 163-169).


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Perhaps the administration of SOD in the form of a heparin-SOD conjugate
(J. Lui c., Mol. Cell. Biochem., 2009, 327 (1-2) 219-228) which
attenuated mRNA expression of TGFi-1 and is thought to be useful for the
treatment of acute liver failure and hepatic fibrosis, should also be considered
of relevance to the possible therapy of ALS.
c
Illnesses where TGFi is thought to be centrally implicated include
rheumatoid arthritis and cancer but could perhaps also include ALS.
The upregulation of the glutamate transporter GCT-1 may occur in the
context of neuroprotective actions of TGFi-like factors.

1-1 Note Relating to the Belinda Cupid (Research Development MND Association England Wales and
Northern Ireland
[Posted on the internet by the Eleanor and Lou Gehrig MDA/ALS Research Center]
Report on the International Symposium, Milan, November 2003
Discussing the use of the mutated human SOD-1 gene in mice, it was noted that ³one of the conundrums of
understanding this form of ALS/MND is why these mutations are only toxic to motor neurons, when they are present
in every cell in the body‰,
[D.G. note: one possibility is that HS is involved in the targeting of this SOD; HS microstructure is believed to differ
for each cell type and HS is known to tether extracellular SOD. Dyshomoestasis of nitric oxide resulting from defects
in antioxidant protection leads to active nitrogen species generation (
. peroxynitrite) and such reactive chemical
moieties could be part of the causative agent cocktails responsible for in tissue damage associated with MNDs and
other degenerative diseases].

To attempt to unravel the specific targeting of the ALS-related

SOD-1 effect it was reported that D. Cleveland had bred mice that containing different proportions of cells that
contained normal and mutated SOD-1 and found that when the amounts of these cells were 35% or more the mice did
not develop ALS/MND, indicating that unmutated SOD-1 can mop up damaging superoxide from neighbouring cells
lacking functionally active SOD-1).
1-2  c#c
c#c
  c#
c& 2
Cytokines involved in nitric oxide regulation are in general subject to
modulation of their activities by HS signaling.


 TGFi
andi (but not i3) activities are potentiated (M. Lyon c.,
J. Biol. Chem., 1997, 27 (29) 18000-18006) by heparin-like HS molecules;
this seems to occur  interference with 2macroglobulin binding.

Numerous academic researchers over the last seventy years have build up an
information database browsing of which can suggest that the heparin/HS-
heparanome, (the system of HS proteoglycansa# which occur in all multi-
cellular animals) is involved widely directly or indirectly in the modulation of
perhaps all animal biochemical systems including a variety direct and indirect
of anti-pathogen actions and furthermore the efficiency of endogenous HS
tissue protection can be simply augmented by (
. dietary) inputs which
seems to promise a revolutionary new approach to science-based healthcare.
HS can be fitted in the centre of a web of influence which radiates to all
animal biochemistry.
An especially potentially important aspect of HS biochemistry is its apparent
involvement with the modulation of nitric oxide actions.
HS is now believed to be the highest level animal biochemistry system
manager which can  ccinterface the genome, the proteome, the
metallome with the environment and so directly affect human and animal
health. This circumstance further suggests that in the future, highly effective
medical interventions will be achievable by clearly defined scientific methods
based on servo feedback alteration of HS microstructures to meet individual
pathological circumstances, so allowing for improved targeted therapeutic
manipulation protocols. This paradigm seems to promise novel therapeutic
methods of highly effectively combating a wide range of illnesses.

The therapeutic use of glucosamine for osteoarthritis stemmed originally from

the discovery of its apparent effectiveness by health food enthusiasts but its
employment as a drug by alternative medicine was not originally well
received by mainstream scientific opinion (›., M.F. McCarty Med. Hypoth.,
1994, 42 (5) 323-327).
It is of some interest, however, in this context, that glucosamine has been
identified much earlier as a potential anti-cancer agent by a highly esteemed
Canadian academic scientist J.H. Quastel, who with A. Cantero (in Nature,
1953, 171, 252-254) reported that glucosamine exhibits significant anti-
tumour activity in a mouse model. (That his finding seems not to have been
followed up could perhaps have been because no credible mechanism of this
action was apparent at that time).
It should be noted that HS is an alternating copolymer of glucosamine and
iduronic or glucuronic uronic acid, so that some of the effects of dietary
glucosamine might be now be attributable to an augmentation of HS
biosynthesis (›., M.F. McCarty, Med. Hypoth., 1998, 51, 11-15).

The possible wide-spectrum therapeutic benefit of the present day (but which
still relatively poorly optimized, 
., as regards which microstructures,
inorganic cofactors ›. which are critically required to enable the specific
bioactivities of heparin-like drugsa#, (
. the list of such drugs must logically
include pentosan polysulfate (PPS)1 ›., as well as glucosamine) can
tentatively be suggested to provide for a major new pro-health concept.

1-2-1 Dysfunctional interactions beween HS and SOD may promote

degenerative disease processes; regulation of heparanase expression (which
gives rise to small HS oligomers) can be achieved by the modulation of the
redox status achievable 

c  the administration of exogenous SOD.
M.L.T. Teoh c
creported their discovery in Cancer Res. 2009, 69, 6355-
6363; doi:10.1158/0008-5472.CAN-09-1195 that the biosynthesis of
heparanase is closely determined by oxidative stress (and related SOD
activity).c
Since heparanase activity can also regulate the production of pro-inflammatory cytokines
and nitric oxide, 
., by T-cells (›., M Bitan c, Diabetes Metab. Res. Rev., 2008, 24
(5) 413-421) the ability of SOD to control heparanase then also further implicates SOD
activity and nitric oxide production.
This indicates that defective SOD (
. as in fALS) will perturb the HS as
well as the nitric oxide signaling systems.
This also leads to neurologically damaging protein misfolding consequent of
pathological nitration (and deactivation of) of protein disulfide isomerase
(PDI).

1-2-1-1 HS Oligomers (or Mimetics Thereof) as Therapeutic Agents

It should also be noted that both nitric oxide metabolites and heparanase actions can depolymerise
HS to yield PPS like molecules which putatively have the abilities to enter into servo feedback
control loops allowing exogenous application of heparin, PPS, ›. to enter into such control loops
to achieve amelioration of a range of tissue damaging impacts.
It should be noted that heparanase action produces HS oligomers which are
approximately mimicked by PPS.
The exogenous application of PPS seems to be uniquely effective for
combating prion diseases, and putatively other illnesses including AD.
Perhaps also ALS.

The above are only two of the many examples which can suggest that unique
health benefits which putatively can to be derived from the HS paradigm.

HS biochemistry can be harnessed to allow the highly effective inhibition of

numerous viral (including HIV/AIDS), bacterial and protozoal infections and
suggest novel, more effective anti-cancer strategies, as well to combat
atherosclerosis.

HS-based therapies promise to show less unwanted side effects than are
apparent with many currently used drugs.

1-2-1-2 Heparanase and Cancer

Cancer mortality is believed eventually by an increased tumor burden
following by tumor metastasis: the virulence of different kinds of tumors in
this respect seem to be correlated with their heparanase activites (›
c
.,
E.A McKenzie, Br. J Pharmacol., 2007, 151 (1) 1-14).
Manipulation of redox status by the administration of SOD can, however, in principle,
inhibit such heparanase-related metastasis (this notion logically follows from the concepts
discussed in the above-mentioned report by M.L.T. Teoh c., of c   studies which
had suggested that low molecular weight heparin (LMWH) administered together with
SOD from which the HS binding site had been deleted (Ec-SOD ˜HBD) is especially
effective at inhibiting tumour cell viability, metastasis and invasiveness).

Augmentation of SOD activity, achieved by the exogenous administration of extracellular Ec SOD attenuated pro-
metastasis heparanase expression (SOD caused an inhibition of the steady state heparanase mRNA was observed c
  which showed that. Interventive manipulation of such SOD actions might then be used to inhibit tumour
metastasis. It was further found that deleting the HS binding residues from extracellular SOD produced a more potent
anti-cancer agent. Since heparanase activity is unexpectedly found to be regulated by oxidative stress this provides
new insight into the dual roles of HS and oxidative/nitrosative stress in degenerative disease processes.

PPS can also act to inhibit cancer-related angiogenesis activities (›., S.

Zaslau c., Amer. J. Surg., 2006, 192 (5) 640-643); this activity was
considered to be of especial relevance to the therapeutic intervention in
prostate cancer.
c
(((c
c  cccc
 c› c c
cc 
 
c
 c
It is of some interest that overexpression of SOD is believed to be an
important part of the etiology of Down¶s syndrome

. the discovery of mutation of SOD was an important breakthrough for the
fuller understanding of the etiology of fALS.

----------------------------------------------------------------------------------------------
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., Multiple Sclerosis (MS) may arise from possible effects of toxic Ba2+ ion perturbation of HS actions
have been proposed, based on epidemiological, data in the etiology of MS.
The binding of FGF-II growth factor to its receptor is thought to require bivalent cations as a essentially
required cofactor; substitution of such cofactors by Li+ and toxic metal ions such as Pb2+, Ba2+ (and
perhaps Al3+) may, however, promote neurodegenerative disease.c., M. Purdey, Med. Hypoth., 2004, 62
(5) 746-754 (›
c
., 2000, 54, 278-306); cf also
A. Visconti c., Ann Ist Super Sanita, 2005 41 (2) 217-22.c
Asbestosis (an Inorganic-Fibre-Induced Disease)
A example how alteration of HS (the principal ligand for extracellular SOD [SOD-3]) can induce the formation of
insoluble fibres arises from studies of toxic asbestos fibre intoxication (
. of the airways which leads to pulmonary
fibrosis and associated cancer). Degradation of HS under these conditions can apparently normally be partly
countered by the presence of SOD, but when HS is also degraded (by the chronic pro-inflammatory actions of
asbestos fibres) this HS no longer binds SOD (›., C.R. Kliment c., Antioxid. Redox Signal, 2008, 10(2) 261-268)
leading to an exacerbation of the inflammatory process.
Gulf -War Syndrome (GWS) is thought to be, at least partly, caused by some kind of environmental-factor-induced
oxidative stress.
One possible toxin is hexachlorbenzene (used as an insecticide, in smokescreen, also formed easily during
combustion processes where any organic molecule is burned in contact with inorganic chloride or organochlorine
containing substance such as polyvinyl chloride, a circumstance which might commonly occur during warfare);
(hexachlorobenzene metabolises to pentachlorphenol, a substance which is known to produce neurological
intoxication (

c›., J. Folch c., NeuroTox., 2009, 30 (3) 451-458).
[
(Insulin growth factor IGF-1) plus Riluzole Therapy for GWS-like Illnesses

., IGF-1 therapy for ALS has been attempted (based on promising results from animal experiments)
There are reports of on-going studies aimed at using IGF-I for the treatment of GWF.
(IGF-1 seems to be reduced in the blood serum of at least a sub-group of such patients).
Perhaps this deficiency of IGF in both ALS and Gulf-War syndrome and related illnesses could further
suggest that similar types of environmental triggers may initiate both disease processes. Studies of the
possible role of depleted uranium induced oxidative and the effectiveness of Riluzole in rat model of Gulf
War Syndrome has been reported to be underway
Y. Watanabe c. reported in Kidney Int. 1992, 41, 1262-1273, that IGF-1 boosted glycosaminoglycan
contents of kidney (especially the amounts of chondroitin and dermatan sulfates present); this research
group also found that serum IGF-1 levels correlated with the cognitive ability of elderly subjects (Dement.
Geriat. Cogn. Disord. 2004, 18, 67-74 [These findings could further suggest that neurotrophic factors,
including those which are implicated in the etiology of ALS may beneficial alter extracellular matrix
glycosaminoglycan (supramolecular structures) thereby greatly upgrading their tissue protective
functions].
It might be suggested that successful IGF-1 treatment for GWS and ALS might be achieved  formation
increased amounts of specific sulfated glycosaminogycans in selected tissues.
It should be noted that ascorbate is known to produce a similar boost in HS sulfation which can be
rationally suggested to be is the likely origin of the Linus Pauling ascorbate dietary supplement method of
combating viral infections and cancer. [ .,D. Grant at web.ukonline.co.uk/dgrant/dg4].
There have been suggestions that aluminium intoxication might also alter zinc biochemistry (
. create
physiological conditions resembling zinc deficiency).
Zinc status has been related to AD (putatively in a HS-determined manner ›., A.J. Busch c., J. Biol
.Chem., 1994, 269 (43) 26618-26621; ›
, also F.M. Corrigan c. BioMetals, 2005, 6 (3) 149-152..
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., aluminium can also directly deactivate superoxide dismutase (SOD). Environmental factors including
aluminium and neurotoxins (
. in drinking water) have been furthermore suspected to have been the cause of the
Guam, ALS-like illness.}.
[
 the first clue as to the origin of familial ALS came with the finding that this was associated with a genetic
alteration in SOD (as discussed below)].
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. mitochondrial Mn SOD becomes deactivated (F. Yamakura c., 
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1998, 273 (23) 14085-14089) following tyrosine 34 nitration.
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c  Protein misfolding induced cnitrosative stress may be the key event in the
etiology of AD and ALS h

This idea that ALS might results from nitrosative stress induced protein misfolding is
strongly confirmed by a recent report (M. Basso c., PLoS ONE, 2009, 4 (12) e8130
doi: 10.1371/journal.pone0008130) which showed that nitration augmented the amount of
detergent insoluble proteins associated with ALS.

The progression of ALS might then be inhibitable by the therapeutic use of anti-nitrants or
inhibitors of inducible nitric oxide synthase (NOS). The usefulness of this idea is
confirmed by a study of an animal model of ALS (K. Ikeda c., J. Neurological Sci.,
1998, 160 (1) 9-15) in which the neuronal NOS inhibitor 7-nitroindazole delayed motor
dysfunction (K. Ikeda c., later reported that oral administration of creatine
monohydrate retarded the progression of motor neuron disease in a mouse model
[Amyotrophic Lateral Sclerosis, 2000 1 (3) 207-212]. (It should be noted, however, that
these finding were not supported by the later researches of F. Facchinetti c., Neurosci.,
1999 90 (4) 1483-1492 who failed to confirm this idea in the SOD defect mouse model of
familial ALS and J.A. Martinez c
c
., 2008, 157 (4) 908-925 also failed to confirm
the any potential benefit from intranasal delivery of insulin and a nitric oxide synthase
inhibitor in the SOD-defect animal model of ALS).
The notion that the biochemistries of nitric oxide and HS are central keys to a fuller
understanding the etiology of ALS is however, supported by later studies reported by
R.M. Mitchell c. (Neurology, 2009, 72, 14-19) which demonstrated that alterations in
the tissue contents of the nitric oxide related cytokines are good biomarkers for the
diagnosis of ALS and, using a list of altered cytokines associated with ALS,
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 which
was the most useful age-related diagnostic.
IL-17 is also known to decrease proteoglycan synthesis (
. putatively upon
overexpression, leading to the diminution of HS tissue protection), but in a   c


 manner (further useful, but less prominent, biomarkers for ALS identified by
Mitchell c. were IL-13 (a suppresser of nitric oxide production and basic fibroblast
growth factor FGF (FGF-II) also a HS-dependent system.

1-1 More on Heparin-Like Drugs

 
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Heparin/HS have a general chaperone functions which can inhibit protein
misfolding and the consequent seeded generation of toxic fibrils therefrom.
This ability may cross-react with how heparin-like molecules can beneficially
modulate nitric oxide biochemistry and allow heparin-like drugs to both
contribute to the induction as well as to the alleviation of nitrosative stress a-1.
The apparently uniquely effective therapeutic potency of heparin-like drugs
for the inhibition of scrapie-like prion infections (including new variant
Creutzfeldt-Jacob disease [nvCJD] ®›
, internet search term ³CJD Alliance‰
accessed files} arises, it can be suggested, at least in part, because of the
ability of heparin-like drugs to simultaneously combat nitrosative stress as
well as to afford protection against a wide ranges of other pathological
insults.
Dual roles of heparin-like molecule signalling related to nitrosative stress in the induction of misfolded protein in the
aetiology of ALS which could suggest that future developments of varieties of heparin-like drugs might in the future
be harnessed to alleviate the circumstances which seem to promote ALS ( !
chronic inflammation induced in
selected cells by the seeded polymerisation of misfolded proteins arising in them by defects in protein disulphide
isomerase (PDI) activity caused by excessive nitrosative stress which leads to the nitration of tyrosine residues
adjacent to the active site of PDI).
Inappropriate amounts of toxic trace metals (
. lead, cadmium and mercury and perhaps aluminium) could
putatively create major perturbation of critical-to-health HS-related tissue protection mechanisms 
. by promoting
the formation of de-N sulphated HS residues (similarly to the alteration in HS microstructure associated with amyloid
precursor protein of AD (›
 I.B. Bruinsma c., Acta Neuropathologica., 2010, 119 (2) 211-220); it also can be
predicted that improved understanding of HS structure and improved methods of sequencing this polysaccharides
might also make possible to µrepair¶ the defective HS in AD by the exogenous application of heparin-like drugs which
seem to enter feedback loops within the heparanome in order to augment a
   biosynthesis of correctly
substituted HS.
Further researches aimed at advancing the application of the above ideas might also be helpful for furthering the
development of more effective therapeutics for ALS a-1.
 c cc c& &cc  c !'c&ccccccc&c
&c cccc c !'c&c&ccc c "
.
[Heparin-like molecules are known to inhibit HIV viral attachment to host cell surfaces and further inhibit of
downstream membrane penetration by a process in which joint HS PDI actions are subducted to achieve viral cell
entry by a mechanism which allows exogenously applied heparin-like drugs to block both mechanisms ®›., R.
Barbouche cl., Molecular Pharmacol., 2005, 67 (4) 1111-1118}].

(HDL-linked) Apolipoprotein E (ApoE) is involved in normal catabolism of

triglyceride-rich lipoproteins; although ApoE4 is thought not to be a
determinant of Alzheimer¶s disease, ApoE variant E4 is the largest known
genetic risk factor for late-onset Alzheimer¶s disease. (a2M genetic alteration
is also a similar risk factor).
Studies by L. Paka c., J. Biol. Chem., 1999, 274 (8) 4816-4823 have
demonstrated how ApoE can combat atherosclerosis  the stimulation of the
biosynthesis of specific anti-plaque heparin-like HS molecules by vascular
endothelial cells; (
. the presence of ApoE signals for increased sulfation
of the HS side chains of the endothelial HS proteoglycans without altering the
core protein production rate). This phenomenon seems to be relevant to how
HDL is a suppressor but low density lipoprotein (LPL) is a promoter of
atherosclerosis. This scenario agrees with the effects of the opposite effects
of HDL and LDL on HS production. LDL was found to reduce HS
proteoglycan (HSPG) synthesis in porcine endothelial cells (B. Olgemoller c
., FEBS Lett., 1990, 264 (1) 37-39) which suggests that elevated LDL
promotes atherosclerosis  the kind of non-denuding blood vessel injury
caused by HSPG depletion.
Exogenous heparin can also boost tissue protective (anti-scale) anticoagulant
HS biosynthesis and therefore putatively can be used a direct method of
combating atherosclerosis. (This function of exogenous heparin was originally
reported to require the presence of an additional ³substance Y‰ by M.A.
Pinhal c., Braz. J. Med. Biol. Res., 1994, 27 (9) 2191-2195; (›. also
Table 3 in the review by D. Grant (2010)
www/scribd.com/doc/26994439/Publication-2-web where some of the reports
which suggest that a variety of dietary and other environmental factors which
can input into altered HS biosynthesis are collected together).
P. Vijayabaskar c., Afr. J. Biochemistry Res., 2008, 2 (5)120-127 have
confirmed in a rat model that exogenously administered standard heparin
produces highly beneficial (statin-like) anti-atherogenic effects. [It should be
noted that possible adverse effect problems have been discussed with statin
therapy (›., B. Golomb c., Drug Safety, 2009 32 (8) 649-661) which may
arise from the induction of oxidative stress which, in small numbers of
vulnerable subgroups, could putatively promote ALS-like illnesses; hence for
such subgroups an alternative use of alternative heparin like lipid status
modifying drugs might be suggested to be more appropriate. [A problem here
is that in general, whereas heparin has been in use for some seventy years, it
can still be argued that there remains an unresolved quality control problem
with this class of drug, 
. unfractionated heparin contains more than a
hundred individual molecules, the amounts of which as well as the associated
inorganic components may vary according to the details of purification
procedures].
An important tissue protective function in the blood circulation system is
however apparently provided by circulating endogenous heparin (., H.
Engelberg, Circulation, 1961, 23, 573; Clin. Appl. Thromb. Hemostasis,
1996, 2 (2) 83-93, andcPharmacol. Rev., 1996, 48, 327-352) which seems, at
least in part, to provide for an up-regulation of the synthesis of endothelium-
located more highly sulfated anticoagulantly-active HS molecules where the
role of µendogenous circulating heparin for limiting the onset of
atherosclerosis is discussed. Endogenous circulating heparin 
., HS
activity, might also similarly inhibit the progression of cancer (›., H.
Engelberg, Cancer (New York) 1999, 85 (23) 257-272) and it might further
be presumed other degenerative diseases. These might include ALS. It
would be worthwhile to attempt to correlate the incidence of ALS with
endogenous heparin status as defined by Engelberg. The role of diet in
augmenting endogenous heparin levels is suggested to be a worthwhile
research topic.
Heparin-like drugs also warrant re-evaluation for AD in the light of the above
scenario since LDL status has now been correlated with the propensity
towards AD.
It should be noted that the HS mimetic PPS has been similarly indicated
provides potential a multi-anti-disease combating drug
(›
capart from inhibiting prion diseases, PPS (Elmiron) appears to be
beneficial and well tolerated when used for the treatment of interstitial cystitis
(V.R. Anderson & C.M. Perry, Drugs, 2006, 66 (6) 821-835); PPS is also
known to inhibit Complement-mediated myocardial injury (this was studied in
a rabbit model bycK.S. Kilgore cl., J. Pharmacol. Exp Ther.,1998, 285,
987-994; ›
 A. Klegeris c., Immunology, 2002, 106 (3) 381-388) (›
,
Complement activation is also believed to negatively impact on the outcome
of neurodegenerative disease processes). PPS is a general mimetic of low
molecular weight heparin which has the ability, unlike unfractionated heparin,
to enter cells. PPS has also been found to inhibit angiogenesis and be a
potential therapeutic agent for prostate cancer (›. Section 1-1-1-2).
----------------------------------------------------------------------------------
1-3
³Ultra low molecular weight heparin‰ (ULMWH)
Low molecular weight heparin, especially ultra low molecular weight heparin
(ULMWH) is believed to be more able to enter cells than is unfractionated
heparin. It should also be noted that ULMWH  c has also been reported
to confer neuroprotection by a similar mechanism to that proposed for lithium
carbonate, namely the modulation of glutamate-induced apoptosis (this
conclusion is however based on studies which were conducted using cortical
cells of rats). It has also been suggested that heparin-like drugs might also
affect a related aspect of mitochondrial activity  a direct effect on
intracellular Ca2+ release (›. T.-G. Yu c., Yonsei Med. J., 2008 49 (3)
486-495; 
., this research group had previously established (›., Pharmacol.
Res., 2007, 56 (4) 350-355) that heparin can protect rats against cerebral
damage caused by ischemia/reperfusion. Such tissue protection seemed to
occur by mechanisms additional to those enabling blood anticoagulation
effects; these may include the inhibition of intracellular Ca2+ release,
antioxidant, anti-inflammatory anti-nitrant, anti -plaque surfactant actions and
other neurotrophic effects.
c
1-3-1 [ 
c" #›
c
ccc$cc 
c c c
Lithium carbonate, studied in animal models of ALS seems to beneficially modulate glutamate activities relevant to
the kind of neurological mitochondrial dysfunction which is thought to contribute to the aetiology of ALS/MND.
Recruitment of ALS/MND patients for a clinical trial of lithium carbonate has been reported to be currently underway in England and
Wales.
A recent Canadian clinical trial of co-administration of the putative anti-ALS drugs, lithium plus Riluzone was,
however terminated, apparently [according to a public announcement] because of a lack of any demonstrable benefit
to patients].

., lithium-(
. as lithium carbonate) finds wide medical use for the treatment of bipolar neurological dysfunction
disorders and these drugs seem to be well tolerated by such patients.
Another form of well-tolerated therapy uses the lithium salt of heparin which as an approved form of anticoagulant
for human use (administered 
., by injection subcutaneously or into the blood stream).
There has been suggestions that oral administration of heparin for pharmaceutical purposes could possibly be
achieved by co-administration of sodium N-[8-(2-hydroxybenzoyl)amino] caprylate which facilitate the gastric
absorption of heparin
(›
 F.G.P. Welt c., Circulation, 2001, 104, 3121-3124).
Fast track trials of this form of such oral lithium administration might be worth considering as potential therapeutic
agent for the treatment of ALS/MND by analogy with how PPS was tested in human subjects for the treatment of
vCJD 2###].

(  ( (c
cc› 
 cc!c 
c
c
c  %c›c  cc
 cc›c
cc  ccc
It is now suggested that such drugs could also be of interest for the development of novel ALS therapies.
The use of the lithium salt of (ULMWH) also be warranted for the alleviation of the symptoms of neurological
diseases in human subjects.
Both anticoagulant and non-anticoagulant lithium heparin but especially ultra low molecular weight forms of heparin
ULMWH) or the heparin-like drug lithium PPS, can be suggested to offer an improved method of delivery of lithium
to appropriate neurological sites of action.

1-3-2 Putative Insulin

Growth Factor-1 (IGF-1)ctherapy for ALS
The report that the progression of MND in a mouse model is greatly inhibited
by an insulin-like growth factor-1 (IGF-1) + heparin-like drug therapy
(A. Gorio c., J. Neurochem., 2002, 81, 194-202)2a could suggest that a
similar benefit might be achievable in human subjects.
The difficulty of adequate delivery may, again, be why IGF-1 alone failed to demonstrate any benefit in ALS patients despite earlier
indications using cellular models, which had indicated that IGF-I promotes motor neuron survival (›
I Nagano c., J Neurolog. Sci.
2005 235 (1-2) 61-68) .

cc
c

-------------------------------------------------------------------------------
(c
ccc  cccccc
c c
)c&c c
cc
*cc
c
(!(c+ ,  c c

(., Glutamate-Related Oxidant Stress)
A glutamate-related activity (oxidative glutathione toxicity) is currently
believed to contribute in a major way to the neuron apoptosis processes
which are apparently associated with ALS. The most-studied pre-oxidant in
this context is glutamate (also thought to be the key chemical agent involved
in the aetiology of ALS/MND) which is thought to block cystine entry to cells
and so inhibit the production of the key antioxidant molecule glutathione
(glutamate is also thought as discussed above to promote mitochondrial
apoptosis and thereby to cause MNDs).
[Glutamate andcc
c 
 of Riluzole (Rilutek®).
This drug is believed to function by reducing the release of glutamate within the nervous system. It is thought to inhibit cell death induced
by the above glutamate associated toxicity mechanism.
A lower-than-predicted effectiveness of this drug has, however, been thought to be caused by a failure of its effective delivery to the
appropriate neurological sites].

( (!(cc)!(
c cc

c
Copaxone (glatiramer acetate, Cop-1) has been reported (D.N. Angelov c
., PNAS USA, 2003, 100 (8) 4790-4795) to protect motor neurons,
including in a mouse model of ALS, against motor neuron degeneration.
It is thought that this intervention counters T-cell-mediated (oxidant and key nitrant) damage. Such damage likely
includes de-N-sulphation of HS.
A high extracellular glutamate presence appears to inhibit the import of
cystine, which results in the depletion of the critical glutathione antioxidant
protection system. This is thought to be part of the mechanism by which cell
death is associated with (
., N-methyl-D-aspartic acid NMDA) receptor-
initiated excitotoxicity (›. D. Schubert & D. Piasecki, J. Neurosci., 2001, 21
(19) 7455-7462).
It is conceivable that improved delivery of IGF-1, Riluzole and other drugs (especially by the co-administration of
heparin) might improve their therapeutic effectiveness for ALS/MND and other diseases 2##.
-----------------------------------------------------------------------------------------
------------------------------------------------------------------------------------------

3 Unconventional Ideas with Supposed Therapeutic Potential

K!(c)&!-c c
Possible Role of Cerium in the Aetiolgoy of Neurological Diseases
(Could Suggest a Li/Ce-heparinate Therapy for ALS/MNDs).

. cerium occurs in human bone.
Nanoparticles consisting of either cerium or yttrium oxide were found to protect rat nerve cells from oxidative stress
caused by glutamate excess. This type of stress is also thought to be the ultimate cause of ALS/MND.

D. Schubert c. (Biochem. Biophys. Res. Commun., 2006, 342 (1) 86-91) observed that synthetic cerium (and
yttrium) oxide containing nanoparticles can confer neuroprotection (in cellular models of nervous tissue) by acting as
an efficient antioxidants (this activity is thought to be dependent on the presence of a mixed oxidation state Ce4+/Ce3+
in the cerium oxide nanoparticles).

Evidence for the requirement for unconventional inorganic elements to enable normal neurological activity to occur
comes from the observed difference in the amounts of cerium in autopsy samples of human brains of patients who
suffered from neurological dysfunctions compared with normal controls.
Cerium may be depleted in those dysfunctional neurological tissues which also benefit lithium therapy (and which
have also been suggested to occur because of a disturbance in cellular redox balance so as to promote the presence of
reactive oxygen free radicals).
(Partly unpublished work from a former Aberdeen University research group suggests that cerium nanoparticles may
occur naturally in association with heparin-like molecules [which appear to bind cerium ions more strongly than other
inorganic ions]; that these exist c   is not out of the question, but requires further research to confirm).c
c
K!(c
&cc .c c

& containing molecules are known to have a prime importance for
antioxidant defence system since glutathione peroxidase activity depends on
the presence of selenium; (

this enzyme is used for recycling glutathione,
a prime antioxidant in animal cells). Impairment of the synthesis of
glutathione peroxidase (under conditions of selenium deficiency) could
therefore conceivably promote ALS/MND.
Selenium dietary supplementation might therefore potentially inhibit these
diseases. A critical review of the information available on vitamin E and
selenium (and other antioxidants) supplementation for ALS, however,
appeared to show little statistically relevant benefits (R. Orrell, University
Coll. London). Other reviewers seem to disagree with this conclusion,
however, and perhaps the overall conclusion form the available literature is
that dietary antioxidant and other vitamin supplementation is beneficial to
ALS/MND patients.
A high profile patient whose disease seems to have remained stable for some
forty years is the physicist Stephen Hawking who is reported to take several
vitamins and minerals each day which he apparently believes to be
responsible for helping to successfully manage his symptoms.
[An AD patient, Louis Blank discussed what seemed to be similar scenario relating to AD in [c
! &c
c
c' 
(].
If the current findings (Solomon c
c›
c›.) relating to µbad¶cholesterol throughout-life effects
promotion of AD are correct and the further correlation between such µbad cholesterol¶ and
endogenous blood heparin status (as indicated by H. Engelberg, ›
c›.) is also a general correctly
identified phenomenon for the general human population, then exogenous heparin administration
should tend to counter AD. Exogenous heparin equivalents might further be afforded by ingestion
of dietary factors such as ascorbate and good lipids which promote blood vessel wall heparin-like
molecule synthesis. This procedure is obviously much more convenient rather than by intravenous
administration or related procedures. Studies of blood heparin should be conducted from the
perspective of confirming the above scenario by the collection of epidemiological evidence for the
existence in human population of interactions between HS status and AD as well as ALS (and
other illnesses).

3.3 A Selenium-Containing Antioxidant which might be useful for the treatment of MNDs ›. :
c(2phenyl-benzo[d]isoselenazol-3-one)
(a hetero-ring selenium containing molecule) of an of approximately similar
chemical structure to Riluzole;
(
. this is also hetero-ring but now sulphur atom containing molecule)
might be useful as an ALS therapeutic agent. This was suggested by
preliminary tests using the mutated SOD-1 mice model of ALS for screening drugs licensed
by the FDA in a program during which 75% of these drugs were investigated. Ebselen appeared to affect
glutamate uptake via the glutamate transporter but perhaps its additional mitochondrial targeted tissue
protective antioxidant/antinitrant properties, which arise from the unusual chemical structure of this
substance, might also offer additional neurological protective pathways of putative benefit to ALS/MNS
patients.

c
c
/cccccc% ccc
& c
c#
*c%.c c
c
c
/!(c c
&c) c
Ascorbate very efficiently reverses the oxidation of nitric oxide to nitrous acid
(nitrite) and therefore is a (major) anti-nitrant.
Ascorbate is also known to upregulate HS tissue protection (by increasing

  biosynthesis of more highly sulphated HS molecules.
(The use of hydroxyurea and arginine as therapeutic agents (mentioned below) could have similar re-balancing effects
on nitric oxide redox dependent balance and therefore also indirectly beneficially modulate HS tissue protection).
Antinitrant action could underpin the key tissue protective role of ascorbate
and be the reason why a dietary supplementation with ascorbate or a diet rich
in fruit and vegetables can apparently slow down the development of AD and
other disease processes
(›., D.K. Lahiri, J. Alzheimers Dis., 2006, 10 (4) 359-361).
The major damaging agent responsible for nitrosative damage/de-polymerisation of HS seems most likely to be (not
peroxynitrite) but nitrite or another derivative of nitrite (
. nitrite is the major primary metabolite of nitric oxide.

R.E.Vilar c., Biochem. J., 1997, 324, 473-497 showed that nitrite can de-polymerise HS. This finding was
contrary to conventional belief that nitrosative scission of HS cannot be accomplished under physiological conditions
at pH 7.4 (previously it being thought that non-physiological acid pH values were required); evidence for the further
requirement of redox metal ions for this nitrosative HS scission process includes the results reported by Vilar c.,
›
c›., who found that the reaction occurred under physiological conditions in the presence of a phosphate buffer
[which is known to contain traces of iron and copper impurities] but not when the reaction was carried out in the
presence of an imidazole buffer). [Cf., A Becaria c., J. Neuroimmunol., 2006, 176 16-23 noted that Al and Cu
ions in drinking water could possibly cause enhanced inflammation and oxidative stress in the brain].

. ascorbate and other natural antioxidants putatively also more generally counter a range of
neurodegenerative and auto-immune disease processes.
(. D. Grant ³Ascorbate and Cancer‰ and
http://web.ukonline.co.uk/dgrant/dg4/
and related sites where the dg4 is replace by dg5 and dg8 (also dg1, dg2 and dg3) reached 

c  search term ³Masler heparan‰ click on ³References‰ which is
µdg8¶, then alter to dg4 ›
in the address box.

More direct evidence for the possible role of nitric oxide balance in neurodegenerative disease process is
that the formation of nitric oxide by astrocytes has been suggested to contribute to the impairment of
mitochondrial functions which lead directly to neurodegenerative processes (

c›., S.J.R. Heales c.,
Biochim. Biophys. Acta-Bioenergetics 1999, 1410 (2) 215-228).
c
c
/!(c cc
&cc0  c ccEFcKccHcc c
cc&cccc!&cc cccK!
 ccc c ccccc'c!1 cc
K!c ccc2!cK!  ccc
cc cc
This suggests that a prime role of h-tocopherol could be to provide antinitrant
protection (., K.S. Williamson c., Nitric Oxide, 2002, 6 (2) 221-227
who reported that the nitration product 5-nitrohtocopherol is increased in the
Alzheimer brain)
During studies (reported by R.J. Singh c., PNAS, USA, 1998, 95 (22)
12912-12917) which had suggested that nitrite provided a suitable probe for
the investigation of the peroxidase activity of the familiar ALS mutated SOD,
nitration of h-tocopherol was observed to occur.
This could suggest that h-tocopherol is the prime lipid-based anti-nitrant
substance.
Hence h-tocopherol dietary supplementation is potentially of major interest
for ALS as well as for AD ›., therapeutic use.
This idea, however, requires further research. [., also K. Hensley c.,
Free Rad. Biol. Med., 2004 36 (1) 1-15)].

 tocopherol (commonly widely available as a dietary supplement which

have been indicated to contain little or none of the K form) had traditionally
been regarded as the most important for tissue antioxidant protection since
this is the believed to be most abundant form present in human tissue;

-tocopherol dietary status or its supplementation in an animal model

seemed, however, not to have any antioxidant protective therapeutic benefit
for AD (as reported by S. Gaedicke c., Br. J. Nutr. 2009, 102 (3) 398-
406).

-+#+'
.V
V/
0
V V
V

Nitrolinoleic is believed to be a highly effective endogenous peroxisome
proliferator-activator receptor (PPAR) gamma ligand.

Nitrolinoleic acid exhibits superoxide generation, degranulation and integrin

expression by human neutrophils (B. Coles c
 Circ. Res., 2002, 91, 375-
381; ›. also, 
., D.G. Lim c., PNAS USA, 2002, 99 (25) 15941-
15945) and potently activates endothelial heme oxygenase (HO-1) expression
(›. M.M. Wright c
c
., 2006, 103 (11) 4299-4304). [HO-1 is
upregulated as an adaptive response to inflammatory stimulae].

Nitration can (perhaps unexpectedly) lead to the formation of tissue

protective nitrated lipids including nitrolinoleic acid. This study of possible
therapeutic effect of this substance and other nitrated lipids against
neurodegenerative diseases is warranted.

-+#+#
V
." 
V 
V

VV

Other dietary components which have been suggested to be of value for the
alleviation of the symptoms of ALS/MND are polyunsaturated fatty acids
(PUFAs) plus Vit. E (J.H. Veldink c., J. Neurol. Neurosurg. Psychiatry,
2007, 78 (4) 367-371.
PUFAS may beneficially augment HS tissue protective functions by boosting
correct HS biosynthesis.
PUFAS might conceivable also diminish chlorinated organic molecule (


HCB) load.

c
/!Kc#.c
This is known to be a useful therapeutic agent which offers benefit to another
disease process induced by the formation of misfolded proteins: sickle cell
anaemia. The 
c 
 of this drug is thought to be  the beneficial
alteration of nitric oxide balance.
It should be noted that hydroxyurea has also been noted to benefit ALS
patients (but modestly).
[. internet site ³Pythagoras‰ ®This is a well stocked scientific scrapbook
with sources of articles about ALS and includes sources of information on the
attempts to discuss the outcome of studies of the use numerous types of drugs
for the treatment of ALS and other diseases including AD} accessed 2/11/09
at
http://www.disabled.gr/forum/archive/index.php/t-11352.html].
This suggests that ALS victims might benefit from other chemically simple
(and inexpensive) therapeutic agents which can be used to affect nitric oxide
biochemistry. These include L-arginine and L-arginine heparin (hydrated)
complexes (
., the delivery of drugs as such complexes is thought to
increase their bio-availability).

4-4c)
(Available as a dietary supplement)

Can Nitric Oxide Brian Tissue Damaging Effects be Countered by Carnosine [i-alanyl-L-histidine] (Perhaps optimally
carnosine + ascorbate + h-tocopherol ?) ?
Carnosine is reported to be a normal component of human brain, and it can further be suggested on general chemical grounds to possibly
function to counter nitrosative stress (including that indicated by the occurrence of neurological tyrosine nitration, an well-established
indicator of the occurrence of nitrosative stress (›., M. Fontana c., Cell. Mol. Life Sci., 2002, 59, 546-557).


 V 



VV


/



V
V



(›
, H. J. Stuerenburg, Biochemistry (Moscow) 2000, 65 (7) 862-865; ›.,
Arch. Gerontol. Geriatr., 29 (2) 107-113).
Carnosine apparently protects against nitrosative stress induced in astrocytes
by lipopolysaccharides and TNF-(›., V. Calabrese c., Neurochem.
Res., 2005 30 (6-7) 797-807) and increases the efficiency of DOPA therapy
in Parkinson¶s disease patients (A. Boldyrev c., Rejuvenation Res., 2008,
11 (4) 821-827).

The origin of degenerative diseases such as ALS might be suggested to partly

arise from environmental intoxication combined with failure in susceptible
individuals of the natural defence mechanisms against such intoxication.

4-5



The possible benefit to ALS patients of tamoxifen therapy was hinted at by B.
Brooks, Madison USA who had found that patients who had ALS as well as
breast cancer, and who were receiving large doses of this drug, seemed to
show a diminished ALS progression which was however not observed in a
cohort of patients who had been treated with a lower dose of tamoxifen.
It might be suggested that 4-hydroxy tamoxifen, a principal metabolite can act
as an effective anti-nitrant as well as being an antioxidant when present in
sufficiently high dose.
Direct use of this (or perhaps another, yet to be identified) metabolite of
tamoxifen is suggested for ALS therapy.
[., H. Wiseman c. (with B. Halliwell), have indicated in Biochem. J.,
1993, 292 (3) 635-638 that cardioprotection is achieved by tamoxifen / 4-
hydroxy tamoxifen by the inhibition of a Cu-ion-dependent lipid peroxidation
process which is thought to occur during oxidative damage to human low-
density lipoprotein LDL (a major mechanism of
promotion of atherosclerosis). Since nitration/nitrosation is now also believed
to be of importance in this disease (›
c

 I Parastatides et al Circulation
Res., 2007, 101, 368-376) protection afforded by 4-hydroxy tamoxifen may
be at least partly due to the anti-nitrant activity of this drug].
Another mechanism by which drugs could afford tissue protective benefit is
by their actions on HS biosynthesis. It should be noted that a beneficial effect
of dietary lipid content can be putatively achieved by this mechanism (›
c
.,
L. Paka c., J. Biol. Chem., 1999, 274 (6) 4816-4823)].

5. Protective Role of
&
This may function to protect against the toxic actions of environmental toxins
including µpesticides¶ of which hexachlorobenzene (HCB) [›
Section 6]
seems to be of especial concern.
Neuromelanin was found to be substantially depleted in patents with
Parkinson disease (›. F.A. Zuccha c., Pigment Cell Res., 2004, 17 (6)
610-617 who discuss the striking affinity of this polymer with metals, lipids,
drugs and pesticides).
A possible correlation has also been suggested between the occurrence of
malignant melanoma and ALS which could implicate a role for the protective
functions afforded by melanin-like polymers in the aetiology of ALS.

5-1

Potential Peroxisome Proliferator Activated Receptor gamma

(..1

) Ligand ±Related Therapy

&&cccc ccc  which
current research suggests such agents could counter inflammatory processes,
improve cognition. These drugs therefore are also of interest for the
treatment of ALS and other neurodegenerative disease processes (›. also, G.
Landreth, Curr. Alzheimer Res., 2007 4 (2) 159-164).
It should also be noted thatc+V

"
V+ "

V 1 (which
can apparently inhibit the progression of AD) are believed to suppress beta
secretase gene promotion activity by the activation of PPARgamma (›., M.
Sastre c., PNAS USA, 2006 103 (2) 443-448).

( cc!c".ccc) c%c

.c
6-1 #. 3 (HCB)
HCB intoxication is known to disturb haemoglobin biosynthesis, induce porphyria and augment
nitrosative stress.
HCB is an abundant environmental contaminant arising from industrial combustion processes,
(
. it seems to abundantly arise from poorly regulated industrial and municipal incinerators); it
becomes transported inter-continentally in the stratosphere and (although the manufacture of this
substance is now prohibited by The Stockholm Convention it has apparently become the principal
dioxin-like endocrine disrupter now present world-wide in human tissue
(including milk ) [The UN sponsored Stockholm Convention does not, however, take account of the
unique route by which HCB is efficiently produced during chlorocarbon pyrolysis c›., N.E.
Aubrey and J.R. Van Wazer, J. Amer. Chem. Soc., 1964, 86, 4380-4383 and D. Grant, J. Appl.
Chem. Biotechnol., 1974, 24, 49-58; a fundamental chemical process which occurs during
numerous industrial processes when organic matter is subjected to pyrolysis (this includes the
incineration of mixtures of molecules containing carbon and chlorine atoms (or choride ions) which
causes HCB to be efficiently µsynthesised¶ or µself-assembled¶ , bio-concentrated up the animal
food chain and stored in adipose tissue]. HCB has been reported (M.D. Stonnard c., Arch.
Toxiciol., 1998, 72 (6) 355-361) to stimulate heme-oxygenase-1 (HO-1) activity and hence this
pollutant can be predicted to perturb nitric oxide biochemistry and thereby, also perturb HS
biochemical processes.
It seems possible that all of the dioxin-like (normally pro-oxidant) environmental toxins will both
create oxidative and nitrosative stress which can progress to ALS and other neurodegenerative
diseases.
[Motor neurons from HO-1-null mice showed elevated levels of nitrated proteins; HO-1 was indicated to provide
front-line defence against nitric oxide toxicity in neuronal cells (A. Bishop c. Biochem. Biophys. Res. Commun.,
2004, 325 (1) 3-9). This indicates that HO-1 activities are probably relevant to the etiology of ALS].

6.1-1
Maternal Intoxication by HCB & the Global Epidemic of Obesity
Obesity is a major world health problem. It is evident that many individuals are affected but not all. It is also evident that genetic factors are implicated. Since this
is a new problem and affects hundreds of millions of individuals it might have been deduced c   that it might be caused by some new generally widely
distributed (most likely by airborne anthropogenic highly toxic particulate chemical substance or substances). A review of the problem drawing on the conventional
biochemical models draws attention to the role of lipostatic mechanisms involving leptins (J.R. Speakman, J. Nutr. 2004, 134, 2090S-2105S) in a wide-ranging
review of the dilemma relating to the scientific explanation of the global obesity epidemic.
The possible role of the HS system as a regulator of lipid turnover is however not generally appreciated outside HS research circles.
The basic knowledge of highly toxic anthropogenic input from combustion of organic substances would again suggest that substances classified as dioxins or dioxin
like substances are prime candidates. The most abundant is hexachorobenzene (HCB). This is the most abundant dioxin-like substance in human milk. Although
HCB industrial production has been banned by the Stockholm Convention it is still being produced as a byproduct of other chemical manufacturing and processing.
The knowledge is there to prevent this but there may either be ignorance of relevant science by lawmakers or effective counterarguments forwarded by industrial
interests.
It now has been confirmed that maternal intoxication by HCB can apparently subtly alter foetal development to pre-dispose offspring to obesity (›., A. Smink
c., Acta Paediatr., 2008, 97 (10) 1465-1469).
This new epidemiological finding promises to account in an intellectual and scientifically sound manner for the current global epidemic of childhood obesity
[and also the inexplicably increasing incidence of AD and ALS which are both linked to obesity and also to intoxication by inflammatory agents; studies by R
Whitmer c. (BMJ 2005 doi:10.1136/bmj.38446.466238.EO) indicated that obesity in middle age increases the risk of future dementia independently of
comorbid conditions].
[Other mystery illnesses including chronic fatigue syndrome (CFS) could also be a consequence of global intoxication by HCB (›
D. Grant file accessed 
browser search term ³Amey Bishop hexachlorobenzene‰)].
On the other hand, arguing against any other than minor input from such an environmental toxin as HCB, a population survey by J Wardle (vide infra) suggested
that genetic factors were almost wholly involved in obesity in general.
6,1-1-1 While genetic factors were argued to be responsible for some seventy percent or more of the variance in weight and body mass index in a large scale
population study of identical and non-identical twins raised in the same or separate childhood environments and therefore environmental factors such as
pollution could not be responsible for the current obesity epidemic (›., J. Wardle c., Amer. J. Clin. Nutr., 87 (2) 398-408).
There is however disagreement about how these findings should be interpreted. Dr. Helen Wallace of GeneWatch UK has been quoted (BBC News Feb. 2008,
internet) as saying that ³twin study findings depend on the assumption that one made about how genes and lifestyle conclusions about the importance of genetic
effects.

It is impossible to draw reliable conclusions from twin data alone‰.

6.1-1-2 Genes need HS to get to work. Studies of twins suggest a dominant role of genetics but this seems at odds with other facts.
The argument about the possible dominance of genes to the current obesity epidemic seems to be quite simply faulted by a consideration that there is a high
likelihood that the human genes were the same some thirty years previously prior to the general increase in childhood obesity.
There is also evidence that modern children take similar dietary intake in calorific terms as their thirty years ago counterparts (›
 ³we have found that people
have not reduced their energy expenditure over the same periods that obesities have increased enormously (Mail Online article 1024159 [interview with Carol
Sparling (believed to be a colleague of Professor J. Speakman, University of Aberdeen who is apparently wrongly named as the source of this article]); modern
children take similar amounts of exercise to children some thirty years. However they may overeat. (This however is also perhaps questionable).
The solution to the enigma surely is that while the genes are the same the genetic expression, RNA processing or HS growth factor control of embryogenesis
(
. that involved in brain development has been affected downstream of the genome by the causative environmental factor(s), so what must be driving the
obesity in the modern child is environmental including global atmospheric contamination. This notion seems to be supported by animal studies. The key
ingredient in solving this dilemma can be suggested to be HS biochemistry. Obesity increases the risk of diabetes and this feeds into altered HS biosynthesis
(which is sensitive to the presence of altered glucose)
The role of genes in human health is of course a much wider issue.
The developing ideas of the role of essentiality of co-operation between genetic and HS systems could in the near future require a complete re-thinking of the
earlier ideas that genes alone determine much of human health and that simple genetic engineering can benefit it except in the case where a single gene defect
leading to a defect in a key protein is responsible. A useful slide discussion of the current ³quackery‰ of some apparent mis-use of genetic arguments has been
posted on the internet by H Wallace, ›
 also www.tbioimed.com/content/3/1/31.

('+#
V
 2"
Stored HCB can apparently be eliminated by dietary fish oil
(as indicated by animal studies, ›
c
., K Rozman c., Toxicology 1981,
22 (1) 33-44 and K. Umegaki & S. Ikegai, J. Nutr. Sci. Viraminol. (Tokyo)
1998 44 (2) 301-311)
If HCB is a promoter of ALS, then dietary fish oil might be beneficial for
ALS victims.


7 Combatting Al3+ Intoxication
V
+3
4""
 5
,
Linked HS Al3+ Induced Dysfunction and ALS?

Although sporadic ALS, as well as familiar ALS could be an antioxidant and

nitric oxide dysfunctions involving protein misfolding and fibrillation, there is
also thought to be some contribution to the etiology of ALS from intoxication
by, 
. some toxic metal environmental factor(s).

Intoxication with Al3+concentrations similar to those present in uremic blood

can apparently efficiently inhibit SOD activity, as indicated by c  
experiments reported by R. Shainkin-Kestenbaum c., Clin. Sci., 1989, 77
(5) 463-466; PMID 2582719.

The existence of geographical hotspots for neurodegenerative diseases (and
their reduction with improved environmental awareness) has suggested that
high Al3+ concentrations, combined with low Ca2+ and low Mg2+ presence in
the environment could be involved in the induction of ALS and other
neurodegenerative diseases (
., J. Durlack c., Magnes. Res., 1997, 10
(4) 339-353; ›., also the historical induction of neurodegeneration in kidney
dialysis patients prior to the realisation that this was caused by the small
amounts of Al3+ present in the tap water. (., The Royal Society of
Chemistry special publication No. 73 ³Aluminium in Food and the
Environment‰ Ed. Robert C. Massey and David Taylor, 1988).

The dyshomeostasis of redox metal ions which has been implicated in the
pathogenesis of AD and other diseases may be the reason why complexing
agents capable of reducing the amounts of such metal ions in the brain seem
to have therapeutic usefulness for the treatment of AD (›. E. Ferrada c.,
Neurotoxicology, 2007, 28 (3) 445-449).
Metal chelators (DP-109 and DP-460) were also found to be neuroprotective
in a transgenic mouse model of ALS (S. Petri c., J. Neurochem., 2007,
102 (3) 991-1000). That such chelating agents are apparently (slightly)
effective in slowing the ALS-like illness in such mice could confirm the
possible general usefulness of some kind of chelation therapy for ALS (›. the
possible use of EDTA for this purpose). There are pitfalls here of course
with this procedure involving unknown side-effects of chelation therapy; a
safer way forward may be to use those chelators which are more in tune with
natural molecules present c  , such as the anionic polysaccharides
(including those of the HS family) or the commercial bisphosphonates, which
may also be in this category). Fulvic acid is a natural chelator (derived from
microbial processing of soil organic matter) which has apparently found use
in European folk therapy as well as in oriental and native American medicine.
A problem, however, with this chelator is uncertainty about µquality control¶
aspects of preparations, !. likely batch to batch variation in the chemical
structures present causing a variation in potency.

8.
The apparent
 ccc
c cccc
(›. Air Line Pilot March 2001 p.16) is something of a mystery, but might be
due to the greater exposure of pilots than to the general population, to
electromagnetic fields.
This accords with the ability of such fields to induce mitochondrial apoptosis.
It is known that pulsed electromagnetic fields can induce apoptosis (this is of
current interest for cancer therapy using high energy pulsed electromagnetic
fields).
The key role of "i 6 V
V

in mitochondrial activity (an activity which is believed to become dysfunctional in ALS) might suggest
that ALS patients should be shielded from excessive electromagnetic energy
(
. especially from (older) mobile phones/ analogue systems).
The polyihydroxybutyrate complex with inorganic polyphosphate shows up at membranes throughout all speciesj and
in animals seems to be important for mitochondrial function and this is now postulated to likely be of major relevance
to the aetiology of ALS/MND. Perhaps the unusual physical chemistry of polyihydroxybutyrate is of relevance in
this context. This was of interest to industrial chemists as it was found easy to manufacture in quantity by
biotechnological means. This polyether was found to possesses piezoelectric properties (somewhat unusual for a
biopolymer but much more common for inorganic crystals); this suggests a mechanism by which cell surface,
including mitochondrial cell surface ion channels, might interact with electromagnetic fields.

There have been hints that polysaccharides can also provide radiation shields
against damaging high energy particles.
This prompts the idea that a similar polysaccharide-based mechanism might
also be capable of shielding tissue against deleterious effects of low energy
electromagnetic fields.
Defects in such extracellular polysaccharides (

their diminution due to
excessive nitric oxide plus oxidative stress) might, it is suggested, diminish
such protection.
Toxin filtration and abilities of heparin-like molecules to promote the removal
of misfolded proteins from cells are activities were they to become impaired,
would be expected to promote diseases which are promoted by prions,
pathological crystals and heavy metal intoxication. This list of such diseases
could include ALS/MND. This prompts the further corollary that
degenerative diseases may occur as a consequence of defects in
polysaccharide biosynthesis or a pathological augmentation of their de-
polymerization. Such pathological de-polymerization of glucosamine-based
polysaccharide systems can in principle (as suggested by c  
experiments) follow from altered redox status and especially from the
presence of excess amounts of reactive oxygen and perhaps more pertinently
reactive nitrogen agents (
. those derived from nitrite plus unliganded redox
metals

9
Other Mitochondrial Manipulations (
. i hydoxybutryate-related) method for beneficial
intervention in ALS/MND? Is this the basis of some dietary programmes?

., the reported
c4!(cc
) cccAmelioration of MND Symptoms
This method is claimed to be highly successful, ›.,
(http://www.tcmtreatment.com/images/ALS-Report.htm),
but the 
c 
 is difficult-to-understand from the perspective of conventional Western biochemistry,
(but not if it can be shown that the recommended dietary input is the major factor influencing HS or i hydroxybutyrate system).
This procedure points to a possible, perhaps rationally acceptable, simple, and low cost therapeutic approach to MNDs.

9-2
A diet-based therapy method for MNDs was also suggested by a report that

 1 
"
V may protect against the progression of mouse-model-ALS
(Z. Zhao c., BMC Neuroscience 2006 7: 29 doi:10.1186/1471-2202-7-
29).
ALS mice were fed a ketogenic diet based on known formulae for humans. This diet seemed, in the animal model, to
improve the mitochondrial dysfunction associated with ALS. It is thought that a key factor was the augmentation of
D-ihydroxybutyrate at the mitochondrial surface. This substance is also available naturally from the cell surface
store of poly-ihydroxybutryate. Defects in the biosynthesis assembly and depolymerization enzymes involved here
can therefore obviously be suggested to be useful targets for researchers aiming to further the knowledge of the
aetiology of MNDs.
Z. Zhao c., claim that therapeutic dietary input method (by administering elevated amounts of ³ketone bodies‰) was the first to show a
potential dietary therapeutic protocol for treating MNDs. This diet can be suggested to modulate the activity of mitochondrial
polyihydroxybutyrate as well as boosting primary biosynthesis of correct HS.
[The former also depends on the presence of inorganic high molecular weight polyphosphate to create the apoptosis ion channel (suggested
to be the ion conducting molecule of the permeable transition pore which is thought to be susceptible to reactive oxygen moieties, ›. at
mitochondrial surfaces [E. Pavlov c
 (with R.N. Reusch) Biophys. J., 2005 88 (4) 2614-2625].

10 Man-Made Polymer Models of Biological Antioxidant Defence Systems.

An enormous amount of information is available from the commercial use of antioxidants needed as essential
additives for rubber and some plastics 
. polypropylene; a test of the role of antioxidants upon the lifetime of
mouldings was given by a comparison of mouldings with . without antioxidants. That without had completely
disintegrated after storage in the air in the dark but that with a synergistic mixture found to produce an optimum
preservation of this organic polymer had not changed. The presence of antioxidants is essential for meaningful
lifetime both with organic man-made polymers and the polymer controlled biological cellular assemblies of living
organisms.
An important aspect of such protection seems to be the existence of a critical synergy between several mechanisms of
protection. For plastics this needs 
., a hindered phenolic plus a divalent sulphur based metal (especially iron)
chelator. The same principle is likely to apply to biological systems.
This suggests that for design of an antioxidant-based ALS/MND therapy the design of a suitable synergistic mixture
of anti-hydroxyl radial phenolics plus iron chelators may also be essential.
This suggests that the efficiency of such agents as Riluzole might be dramatically improved by the co-
administration of an iron chelator, e.g. that identified by
Kupershmidt c., FASEB J., 2009, 23, 3766-3779

------------------------------------------------------------------------------------

O

V










V
OV

VV
 ! (heparin-like drugs). These are
semi-synthetic mimetic of heparin prepared from marine algae or for SP54
from beech wood shavings,
which can be suggested on the basis of published and unpublished information to allow, especially following further optimization of the
chemical structure of this type of anion (which apparently can enhance the anti-viral HIV-1 activities of the normally obtained mixtures of
PPS-like molecules addendum) the dawn of a new kind of highly effective therapy for a wide range of degenerative diseases (›., N.V. Todd c
., J Infect. 2005, 50, 394-396, ›., I. Zerr, Infectious Disorders Drug Targets, 2009, 9, 92-99 for discussion of preliminary studies of the
possible unique therapeutic benefits which may afforded by this class of drug for the treatment of variant CJD (vCJD) a (Mad Cow
Disease) victims;
in this context it should be noted that the metabolic fate of prions is believed, c   to at least in part, be determined by PPS-like
(heparin-like) substances (›
c

R. Gabizon c., J. Biol. Chem., 1996, 271, 16856-16861, 

 2005, 280, 17057-17061 and V.A.
Lawson c., (Abstr. P29, Proc. III Int. Symp., New Prion Biol.. Venice, Apr., 2009) who confirmed the notion that ³subtle changes in the
glycosaminoglycan repertoire‰(
.heparin like structures) influence the pathogenesis of prion diseases}. That scrapie infection of
neuroblastoma cells precludes nitric oxide production when the cells are challenged with lipoprotein (H. Lindgren c., J. Neurosci. Res.,
2003, 71, 291299; ›., K. Mani c., Glycobiology 2004, 14, 599-603 ) further suggests that the aberrant deaminative cleavage of
heparin-like molecules (which can produce small membrane entering PPS-like entities c  ) may be central to the pathogenesis of
scrapie and other neurodegenerative diseases.

A possible role of Ca PPS in the regulation of proteolysis activities which thought to offer possible cancer
therapeutic benefit, has been indicated by H. P. Sorensen c., J. Biol. Chem., 2008, 283, 31920-31932.

--------- --------- --------- --------- --------- --------- ---------

2
 c#c
c#
c &ccc
&c&

It can be suggested from the published literature in HS in peer reviewed

journals that HS exerts numerous potential pro-health actions.
(›., www.scribd.com/doc/2694439/Publication-2-web)
It should be noted that the link noted between blood µbad¶ cholesterol lipid and HS status could also
underpin the recently discovery (A. Solomon c., Dement. Geriat. Cognit. Disorders, 2009, 75-80) of an
epidemiological link between cholesterol status and AD which can be alternatively be rationalized in
terms of the status (
. the integrity) of endogenous HS-related tissue protection mechanisms.
It should also be noted that, an apparent pro-ALS side effect of statin therapy for atherosclerosis, which is
now apparently coming to light (›., B.A. Golomb c., Drug Safety, 2009, 32 (8) 649-661) could
indicate that manifestations of the underlying dysfunction responsible for these and other degenerative
diseases is some HS biochemistry-defect related process. Such defects seem, however, putatively be
repairable by the exogenous administration of HS-containing drugs, heparin, PPS and perhaps also by
glucosamine [›. M.F. McCarty Med. Hypoth. 1997, 48, 245-251] (and also, putatively, a list of beneficial
dietary components discussed by Pillarisetti ›
c›. which must also include ascorbate, which boost HS
biosynthesis). This unexpected benefit seems to derive from the servo control feedback information
processing nature of the heparanome which apparently employs low molecular weight HS fragments as
signaling units and small molecule dietary components as adjusters of HS biosynthesis. The possibility
also exists of an adverse effect of exogenous toxic substances on HS biosynthesis including those known to
be induced by toxic metals (
. lead which has been proposed to promote cardiovascular disorders via the
negative effect on HS proteoglycan biosynthesis) and putatively also, by certain man-made organic
substances including those which have been employed for human drug use, but the fuller understanding of
such effects requires further research.

The pro and anti-HS proteoglycan effects of pro and anti atherogenic
molecules (›., Pilarisetti ›
c›.) is but one example of an range of similar
indications of how the tissue protective functions of HS are modified by
environmental factors which can enable a rational system of intervention to be
achieved by the modulation of HS-directed positive and negative feedback
stimuli which impact on health. HS seems to be the only biopolymer which
changes in structure according to age in a manner which can be further
suggested to diminish its protective functions (or even engage in tissue
damaging functions). HS can however also be altered by dietary factor input
(
., ascorbate and retinoic acid are beneficial but such inorganic ions as lead
cadmium and mercury as well as fluoride when present in large amounts are
detrimental). It seems that simple regulation of dietary factors can, at least
partly, reverse the natural age-determined diminution of HS tissue protective
function but that the ingestion of toxic lead, mercury and cadmium and
perhaps also aluminium ions could putatively diminish the tissue protective
functions afforded by HS.
HS Cross Talk (Relevant to Tissue Protection and Inhibition of TSEs?)
It was found by L. Jacobbson c. (Dev. Cell, 2006, 10, (5) 625-634) that HS from neighbouring cells can similarly
substitute for defective HS for provision of VEGF growth factor receptor activity.
This surprising discovery was announced in a hyped Uppsala University press release, ³New Discovery About Role of
Sugar in Cell Communication‰ (www.medicalnewstoday.com/articles/43115.php). The academic report, however,
was more muted in its discussion of the relevance of this finding].
D. Cleveland was reported to have mentioned that his finding was of major therapeutic relevance to the design of
ALS therapies as ³the delivery of non-neuronal cells to spinal cords could be completely protective, without replacing
a single neuron‰.
While he was thinking of stem cell based therapy, the boosting of antioxidant defence  HS-based medication,
might also be indicated.

2a
An example of how HS proteoglycans and their polysaccharide-based tissue
regulation and protection functions become disturbed under diabetic
pathological conditions is illustrated by how the kidney filtration function
becomes diminished in diabetes. This apparently occurs  a direct effect of
elevated glucose concentration which feeds back to adversely alter primary
the HS biosynthesis (›., N. Parthasarathy c
Mol. Cell Biochem., 2000,
213, (1-2) 1-9 (
. this seems to be how excess blood glucose in diabetes
diminishes the critical anti-atheroma N-sulphonate group content of HS).
. also Note g

2##
#777
A low molecular weight heparin-minetic PPS1 is known to inhibit
neurological degeneration in transmissible spongiform encephalophathies
(induced by misfolded prions). This conclusion is derived from tests
conducted on human subjects. There is some confusion as to the ability of
this drug to prevent damage already caused by prions prior to the
administration of the drug.
The results suggest that further development of this kind of drug for nvCJD
and other diseases could be warranted.
[The use of this drug in the UK was permitted following legal challenge by a
patient-directed interest group (Graham Steel, Glasgow, former member of
the CJD alliance, personal communications)
›., Roger Highfield, HYPERLINK
http://www.telegraph.co.uk/science/science-news/3313916/Dramatic-results-
of www.telegraph.co.uk/science/science-news/3313916/Dramatic-results-of -CJD-treatment.html
The apparently unique effectiveness of PPS against vCJD may depend on
multiple synergistic activities including, a wide-range ³surfactant‰-like (as in
traditional colloid science) anti-plaque potency of heparin like drugs which
enables them to prevent, by the blocking of rate-determining seeding
processes, the detrimental formation of ³infectious‰ sparingly soluble
inorganic organo-metallic and organic phases created by misfolded proteins.
This scenario could suggest that studies of a directly targeted neurological site
infusion of heparin-like drugs (including PPS) for the therapeutic intervention
in MNDs, could also be warranted].

3
On p. 2 of the document ³Amyotrophic Lateral Sclerosis A Guide for Patients
and Families‰ (2001, available in abbreviated form on the internet) by H.
Mitsumoto & T.L. Munsat) it is noted that the prevalence of this disease is 1-
7/100000 but that clusters with a higher prevalence had been reported in three
islands in the Western Pacific including Guam (however for the past 40 years
new cases have declined steadily in these regions).
[The presence of U.S. military bases in this region and intoxication by metals (
. aluminium as well as highly toxic
beryllium from bearings from the servicing of aircraft and soluble forms of aluminium) getting into drinking water
may be part of the reason for this.
[It has also been suggested that the neurotoxinimethylaminoalanine (a
substance of cyanobacterial origin present in cycad tissues formerly used for
food) can induce an ALS/Parkinsonism].
On p. 5, it is noted that ³Charcot in 1875 may have been the first to question
the relationship between remote poliomyelitis infection and the subsequent
development of ALS«this question remains open‰. It was also mentioned
that: ³Interest in heavy metal intoxication as a cause of ALS has been high
for nearly a century. Wilson in 1907 is credited as being the first to raise the
question as to whether lead poisoning causes ALS‰.
The reviewer concluded however that ³it is likely that heavy metals play only
a marginal role, if any, in the development of this disease‰.
On p.6 the mutation in the superoxide dismutase connection was noted.

It should be noted that B. Bocca c., (Ann. Ist. Super Sanita, 2005 41 (2)
197-203) have confirmed the likelihood of a contribution of Al intoxication in
AD by a study of the amounts of 26 metals in the whole blood and serum in
AD patients . controls. Serum aluminium was also found by N.B. Roberts
c
(in J. Inorg. Biochem., 1996, 69 (3) 171-17) to be significantly raised in
dementia patients including those with AD.
The Birchall hypothesis of aluminium intoxication, that sufficient poly-silicic
acid, present in drinking water or in the diet, prevents aluminium intoxication,
has also been general confirmed by epidemiological studies. An extension of
the Birchall hypothesis can however be suggested: that the silicon in HS plays
a protective role in the overall tissue protective actions of HS.
The ability of silicic acid to deactivate neurotoxic aluminium might suggest
that some form of silicic acid might have therapeutic benefit for ALS victims.

Are all ALS victims genetically pro-disposed to elevated metal

hypersensitivity?
And if so they should obviously avoid exposure to such metals.

It should be noted that it is a commonly held belief that aluminium

intoxication is little concern for the general population (this idea was
supported, 
., by the careful studies of H.-A. Hellstrom c
, (Age and
Ageing, 2008, 37 (2) 217-2200) which reported their finding s that the
aluminium contents of orthopaedic patients¶ bones clearly indicated that the
increased presence of aluminuim during lifetime in this (the principal storage
site) was not associated with any increased mortality.
This finding might, however, indicate that some other anthropgenically
enhanced toxic metal, not aluminium (but perhaps, 
., lead or beryllium, or
lack in the diet of some conventional or some unconventional essential metal
(
. some of the rare earth elements) should now be the focus of attention for
the elucidation of the nature of a metal-related intoxication which could be an
environmental trigger of neurodegeneration.

Heparin has intrinsic SOD activity of interest for the direct ability of heparin
to influence nitric oxide stress, but this may be dependent on the occurrence
in heparin of traces of redox metals. Some heparins are known to contain
appreciable amounts of aluminium (

 Bohrer c., RBAC (Brasil) 2004,
36, 99-103). The report that Al3+ inhibits enzymic SOD activity might also
apply to heparin-like SOD activity; those heparins which contain additional
aluminium would therefore be less active as anti-degenerative disease
therapeutic agents than those which contain less aluminium.
Intrinsic superoxide dismutase (SOD) activities of heparin were reported in a
preliminary report by D. Grant c., Biochem. Soc. Trans., 1988, 16, 1030-
1031; ›., Inflammation Res., 1994, 41 (2) DOI 10.1007/BF01987642). It
should be noted that heparin is also a multi-counterion (multi-element) ligand
(D. Grant, CPS 2000 biochem/001/0002; ›., D. Grant, HYPERLINK
http://www.scribd.com/doc/26994439/Publication-2-web
www.scribd.com/doc/26994439/Publication-2-web (2010)) so that different batches
might contain different amounts of (SOD-activity promoting) metal ions;
some of these could be essential to create a suitable therapeutic agent to
combat MNDs. To upgrade this concept into useful therapeutic agents,
further research is required.







------------------------------------------------------------------------------------------
4
 V
  V
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8

It should be noted that





VVV

V

which can occur in renal insufficiency in experimental animals has been associated with nitrosative stress
and tyrosine nitration (G. Deng c
c›
c›.)
V


 

V
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VV

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V



 V
"



V


"

V

V
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$%V

V (while normally this form of SOD occurs extracellularly, in the brain SOD-3 apparently also has an
intercellular function).
That this mechanism could be relevant to the role of nitrosative stress in the aetiologies of
brain dysfunction diseases is indicated by the generation of impaired memory and
enhanced injury in stroke models in SOD-3-null mice (›., S.L. Marklund in
 
  c Ed. M. Flint Beal c., Cambridge University Press, 2005).

Overexpression of SOD-1 can, perhaps unexpectedly, also lead to oxidative and probably more
importantly, nitrosative stress.
Experiments with cultured neurons from transgenic Cu/Zn SOD mice,
producing elevated Cu/Zn SOD activity. showed higher susceptibility than
did non-transgenic neurons to kainic acid mediated excitotoxicity (O. Bar-
Peled c., PNAS, USA, 1996, 93(16) 8530-8535).
There is an apparent need for an exact level of SOD activity, as this protein is believed to enter
feedback loops which involve other antioxidant systems, so that if SOD becomes elevated, these
other systems become impaired. This scenario involving the over-expression of SOD-1 was
originally thought to occur in Down¶s syndrome where it generated oxidative stress (the apparent
origin of AD-like symptoms) but later work to test the prime role of dysequilibrium of SOD and
glutathione in Down¶s syndrome affected and normal siblings did not substantiate these earlier
suggestions that this disease arose from a defect in the antioxidant enzyme activities (›., P.M.
Neves c., Down Syndrome Res. Practice, 2002, 8 (2) 79-82).

The putative biochemical association of misfolded SOD-1 dysfunction with

the occurrence of ALS can also be relevant to the fuller understanding of how
neurodegenerative diseases can be promoted in diabetes (
. the key PDI
protein can cause reductive degradation of insulin; diabetes is also known to
cause,  glucose effects, a direct primary biosynthetic alteration in the
microstructure of HS)g.
--------------------------------------------------------------------------------

, 

 

-  

Footnote a
., articles by D. Grant c. listed at HYPERLINK
"http://www/abdn.ac.uk/~bch118/publications2003.doc"
http://www/abdn.ac.uk/~bch118/publications2003.doc ; cc
  c
›c›c%cc
cc%c
cc ›c›c µ) c
&cccc c
c›  c›c c›

ccccc
c
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+
c&c %
c
. also www/scribd.com/doc/26994439/Publication-2-web.

This document arose out of ideas gained during the nineteen eighties and first the part of the nineties during researches commenced
in 1975 (with Professor J. Tinsley) at the Department of Soil Science, Kings College Aberdeen and from 1981 in a biochemical
polysaccharide research laboratory at Marischal College, Aberdeen and by knowledge gained during personal employment as a research
chemist (including at International Synthetic Rubber Co Ltd., Hampshire, UK) where knowledge was gained on the ability of phenolic plus
bivalent sulphur molecules to synergistically confer antioxidant protection and controlled lifetimes on man-made organic polymers. Earlier,
during a tenure of a research fellowship with J.R. Van Wazer (who headed an academic style international-recruited open-information-
publishing laboratory hosted by Monsanto Hq., 800 North Lindberg Bvd., St Louis, USA) it was observed that divalent sulphur-sulphur
clusters seemed to engage in self assembly (

cstarting with organic disulphides); this was suggested by the initially produced NMR
spectra of organic disulphide mixtures briefly mentioned in the J. Amer. Chem. Soc., 1964, 86, 3012-3017 article which described the
subsequent structural reorganisation behaviour of bivalent sulphur monomers and polymers  S-S bond interchange.
D. Grant c. later noted in Med Hypoth., 1989, 28, 245-253 in an article entitiled [c› ccc
c ' cc
 cccc›c ccc
c›c c› cc ›c
c c ›c
c›
c 
( that a wide range of proteins seemed capable of forming the suggested types of through space divalent sulphur based
clusters including putative examples where the bivalent were juxtaposed to aromatic residues, especially to tyrosines, for which their
occurrence and variation in protein amino acid sequences and the conservation between species throughout evolution, suggested the need
for the attainment by biota of a similar antioxidant protection system to that which had been independently arrived at by industrial chemists
for the preservation of 
. polypropylene man-made polymers in oxygen-containing environments. In the final paragraph of this article,
protein disulphide isomerase (PDI) is briefly noted to be an example of the highlighted type of cysteine tyrosine structure. The general
cysteine tyrosine system was later suggested (D. Grant and F.B. Williamson, unpublished) to be involved in nitric oxide metabolite control

. for the inhibition of unwanted protein nitration by providing a scavenging system for the active nitric oxide metabolites which are
responsible for this. This suggests that proteins of the type highlighted in this article might potentially be able to protect against the
suggested principal disease promoting pathway in ALS. This is especially because t

"V V
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V"1 
While our original idea was that the cysteine-aromatic cluster system afforded the traditional kind of antioxidant protection (but wider and
of a more general nature than the accepted textbook systems animal antioxidant protection provided by superoxide dismutatse, catalyse,
glutathione, ›.) later studies and discussions extended the notion of proposed antioxidant tissue-protective reactivity of cysteine-tyrosine
clusters to include activities relevant to nitrosative stress. (This included laboratory researches conducted in support of the postgraduate
researches of Gillian Mackintosh (Ph.D. 1995) into iron heparin and heparin-like molecule interactions and extension of these ideas with
Jeanette Grant, Prof. K.E.L. McColl and colleagues at Glasgow University in 1999 (›., http://web.ukonline.co.uk/dgrant/dg2/) and later
during debates with F.B. Williamson and V. Spence (in connection with The Royal Society of Edinburgh/Wellcome Trust Workshop ³
New developments in the biology of Myalgic Encephalomyelitis/Chronic Fatigue Syndrome held at the West park Conference Centre,
University of Dundee on 3rd October, 2003) and with Graham Steel (formerly of the CJD Alliance) relating to the
mechanisms by which heparin-like molecules offered therapeutic potency for the inhibition of prion diseases
(including in humans).
A further possible relevance of the antinitrant/nitrosative stress hypothesis is that it might be relevant to the aetiology
of HIV-AIDS which is currently thought to arise  viral attachment to host cell heparan sulfate and another heparan
related PDI-dependent mechanism of cellular entry. It should be noted that heparin-like drugs are highly effective at
combating HIV infections (putatively by a range of mechanisms including interaction of heparan sulphate and PDI
actions).
$
Unpublished researches from Marischal College Aberdeen, contributed to by the author, had also confirmed by c   assays of that HIV
is inhibited to a comparable degree of efficiency to AZT by heparin and heparin-like substances including pentosan polysulfate (the specific
drug which also uniquely seems able to inhibit prion diseases). Studies which seem of relevance to this scenario include investigations of
the physiochemical interactions (including metal ion binding behaviour) of beech wood prepared pentosan polysulfate as well as the
preparation of µimproved forms¶of pentosan polysulfate from other sources; that from a marine algal source was found to be a uniquely
highly potent inhibitor of HIV viral infection in an c   cell model studies and might therefore might also be useful for future researches
into the design of improved therapeutic intervention procedures for prion and other protein misfolding diseases, including ALS and AD).
Extensive studies of how Al3+ ions bind to heparin also conducted by the author indicated that the production of an acidic buffer system
observed c   could if also produced c   could be of relevance to the fuller understanding of the mechanism of toxicity of Al3+.



Heparin-like drugs can inhibit TSEs. This, at least in part, could depend on a general
ability of heparin-like segments in heparan sulfate to specifically inhibit the formation the
infectious prion infective agent aggregates; the formation of such aggregates seems to
occur by a mechanism akin to how these polysaccharides can non-enzymically peptize
fibrin or hydroxyaptatite a behaviour which seems to be utilised by animals to control
pathological inorganic solid phase formation at blood and urinary vessel walls (N.R.
Deleault c., PNAS USA, 2007, 104 (23) 9741-9746; ›
 also, the possible role of iron
ion determined aggregate formation K. Nishina c., J. Biol. Chem., 2004, 279, 40788-
40794; cthe inorganic plaque inhibitory activity of heparin-like substances, which may
have relevance for a fuller understanding of the mechanism of the inhibition of prion-
inorganic adduct aggregates by heparin-like substances was also discussed by D. Grant c
., Med. Hypoth., 1992 38 49-55).c
It should be noted that healthy animal organisms seem to have the ability to adapt to threats from common
hazardous such as calcium oxalate crystals  the ability of nascent crystals to signal for the biosynthesis
of specific heparan sulpfate molecules to stop pathological crystals from being formed from such seeds,
›., F.T. Borges c., Kidney Int., 2005, 68, 1630-1642).
This could suggest that in general the formation pathological relevant solids especially those with a high
inorganic content is dependent on the efficiency of the heparin/heparan sulfate anti-plaque mechanism.


7
Originally conceived as simply being scaffolding in the extracellular matrix,
HS polysaccharides are now viewed as high ranking systemic directors of
animal biological activities; this includes development (
. enabled by the
dynamic biosynthesis of HS sequences in wound healing and in the
development organs [including the brain] and organisms) (The heparanome
(›., S. Guimond c., Biochem. Soc Trans., 2002, 29 (2) 177-181; PMID:
11356149) is a new concept in which dynamic changes in biosynthesis of
different HS sequences [which contain information processor provide a bar-
code-like signals which are modulated by essentially required inorganic
cofactors ›
c
. T.R. Rudd c., Carbohydr. Res. 2008, 343, 2184-2193;
Glycobiology, 2007, 17 (9) 983- 993; 
., 2009, 19 (10 52-57 ] create
distinct cellular HS repertoires which occur in specific spatio-temporal
patterns; this endows organisms with a master regulatory mechanism which
directs the activities of the vast number of HS binding proteins as well as
critically influencing inorganic biochemistry. This HS servo feedback system
process seems also to enable evolution, there being likely analogous
polysaccharide-based systems in other multi-cellular organisms. While in no
way downgrading the centrality of nucleic acid information storage (DNA and
RNA) to life processes, the heparanome seems essentially now, especially in
the post-human-genome era, to possibly be about to replace the genome as
the principal focus for frontline biochemical researches.
While eventually it might be hoped that directed alteration of specific HS
signals might be harnessed for therapeutic purposes, at the present time the
usefulness of heparin-like polysaccharide-based drugs more depends upon
(except for the widespread medical use of heparin as an anticoagulant for
control of blood coagulation) upon relatively poorly understood wide-ranging
tissue protective roles which include µprimitive¶ anti-nitrant and antioxidant
actions as well as the general ability to inhibit the formation of the kinds of
solid phases which occur in amyloidoses (

AD and vCJD, and putatively,
also, ALS). [
, heparin-like molecules have been confirmed by independent
investigators to inhibit amyloidosis in cellular models thought to be relevant
to Alzheimer¶s disease (R. Kisilevsky c., Nature, Medicine, 1995, 1, 143-
148; ›., FASEB J., express article 10.1098/fj.03-1436fje, 2004 (E. Elimova
c.) and H. Zhu c
 Mol. Med., 2001, 7 517-522) and low molecular
weight heparin was found to reduce plaques and F-amyloid accumulation in a
mouse model of AD (L. Bergamaschini c., J. Neuroscoi., 2004, 24 (17)
4181-4186).

It should be noted that HS also promotes superoxide dismutase (SOD-3)

endothelial wall attachment, dysfunctions of which have been implicated in
the aetiologies of a range of degenerative diseases including atherosclerosis,
AD and in the human form of mad cow disease, (vCJD) and as is now
pointed out also are implicated in ALS. All of these diseases are additionally
thought to arise from damaging effects of misfolded protein aggregates which
are in part at least promoted by redox system dysequilibrium and metal ion
intoxication.
It seems evident that HS biochemistry may centrally impact on vCJD,
atherosclerosis (the published research information on HS now suggests that
atherogenic molecules negatively regulate HS but, on the other hand,
antiatherogenic molecules positively regulate HS (›., S. Pillarisetti, Trends
Cardiovasc. Med. 2000, 10 (2) 60-65).
This circumstance needs, however, to be more fully brought to the attention
of the wider µscientifically literate community¶ to, 

 stimulate discussion of
what could be related mechanisms by which pentosan polysufate (PPS1 
.
SP54), a heparin-and HS-fragment-like drug, inhibits neurodegenerative
transmissible spongiform encephalopathy (TSE) diseases, including the
human form of mad cow disease, new variant Creutzfeldt Jacob disease
(nvCJD).

A direct trial of heparin-like drugs to test their potential abilities to inhibit

misfolded protein formation using, 
., the kinds of c   seeded
crystallisation procedures as was described by D. Grant in Biochem. J., 1989,
259, 41-45, is perhaps warranted.
(Not all types of heparin and heparin-like drugs might be active for this effect
however, this activity may vary with the inorganic 
. SiO2 content of
heparins).
This approach might allow the rapid screening of potential drugs for possible
use as therapeutics for ALS, AD and other diseases.
(
., L Bergamaschini c. (›
c›.) have indicated that low molecular
weight heparin reduces plaque formation in an animal model of AD; trials of
this kind of drug with AD patients seem already to have been conducted some
two decades ago with apparently promising results having being reported 
.
for the heparin-like drug Ateroid, which as suggested by Eur. Pat. 0293974
apparently underwent a successful preliminary clinical trial for AD therapy].

Footnote a-1
Excessive nitrosative stress could be the principal origin of the pathological insoluble inclusions which
eventually kill affected neurons.
There are hints (›., G.R. Upchurch c., J. Cardiovascular Pharmacol. Ther., 2001, 6 (2) 163-173).that
heparin can inactivate nitric oxide and eliminate toxic nitric oxide metabolites and hence counter
nitrosative stress.
Excessive nitrosative stress is apparently associated with ALS which may arise mainly as a consequence
of caused by nitration/ nitrosylation processes which include the deactivation of protein disulphide
isomerase (PDI). When tyrosines adjacent to the active S-S isomerae site become nitrated, they render
PDI inactive for the prevention of protein misfolding and hence such nitration promotes the kind of fibril
and plaque formation which is known to be associated with neurodegenerative diseases including ALS.
This excessive nitration may occur following eficient endogenous heparin-like tissue protection. This
critical health-promoting action was identified by H. Engelbert (›
›,
ccc
PDI is also thought to act as a reservoir of nitric oxide which resemble the nitric oxide reservoir system
associated with the conserved cysteines in heparan sulphate core proteins which are believed to release
their nitric oxide in a zinc and copper ion plus ascorbate determined manner to provide heparan sulphate
oligomers which are thought to enter into servo feedback signalling systems which can assist with the
tissue protection mechanisms but which if defective, promote numerous degenerative disease processes.
Exogenously applied heparin-like molecules seem to by entering into such servo control loops, allow for
beneficial intervention to be achieved in degenerative disease processes.

Heparin-like polysaccharides also provide potent anti-viral actions (including those which are
neuroprotective). Heparin like drugs are known to be especially effective in combating AIDS disease
processes. While heparin inhibits key proteolytic and reverse transcriptase enzymes this drug may also
provide the optimal therapeutic approach to combating full AIDS since heparin can apparently inhibit a
range of other interactions which are thought to be responsible for AIDS. These include the ability of
heparin-like drugs to block the initial HIV viral attachment to host heparan sulphates at target cell
surfaces as well as the ability of heparin-like molecules to block subsequent endocytosis  viral
subduction of PDI-cell surface heparan sulphate cooperative interactions (in which reduction of HIV Env
by PDI appears to involve critical control actions of adjacent heparan sulphate molecules; ›., R
Barbouche c. Mol. Pharmacol., 2005, 67 (4) 1111-1118.

It is now suggested that heparin-like drugs might also be capable of blocking ALS/MND by utilising a
similar battery of protective mechanisms including PDI anti-nitration protection..

It should also be noted that heparin-like drugs are also putatively able to combat chemical hypersensitivity
(e.g. arising from the actions of 
. Al3+ Pb2+, Hg2 and Be2 and Fe3+ ions which potentially are able to
augment nitrosative and oxidant stress).
It should be noted has been reported that 12 out of 13 ALS patients tested positively for enhanced
lymphocyte responses to a range of metals (J. Stejsakal & V.D.M. Stejskal, Neuroendoc. Lett., 1999, 20,
351-364) which suggests that ALS could be, at least partly, a metal-induced hypersensitivity disorder, at
least in these 12 patients.
[The insufficiency of dietary inorganic SiO2 sols content in diet may also deplete natural polyuronate cell
surface and extracellular matrix protection systems which can putatively function to deactivate by
engulfment such potentially toxic metal ions].

Hexachlorobenzene (HCB), which can be classed as a dioxin-like substance which is now widely present in human an
animal populations, seems to induce abnormal nitrosative stress. HCB also induces the natural opponent of
nitrosative stress, the antioxidant and heat shock protein heme oxygenase±1 (HO-1) which inhibits protein nitration.
ALS like numerous other degenerative disease processes has been associated with the occurrence of nitrated proteins
in the affected tissues.

Heme oxygenase-1 (HO-1) (and an antioxidant heat shock protein HO-1 activity seems to be related to the induction
of ALS. Induced adaptive resistance (IAR) to nitric oxide in the central nervous system is apparently related to the
ability of HO-1 to inhibit protein nitration.
a-1-1
It should be noted that these properties arise from the kinds of colloid-type
phase boundary surfactant actions which had been the subject of intensive
studies by classical colloid and capillary chemists many years ago e; such
classical researches showed that the physical chemistry of polyanionic
molecules can be subducted to either accelerate or inhibit phase boundary
processes which potentially include the aggregation of misfolded proteins;
this behaviour is an example of a relatively primitive µabiotic¶ function which
seems to have evolved further in multicellular animals to create the currently
used system which affords a µsmart¶ regulation of selective protein
conformation change which is most commonly exemplified by the highly
specific heparin pentasaccharide antithrombin complexation process, a key
event in the mechanism of heparin anticoagulation action.

It has been indicated (R.I. Dima & D. Thirumali, Protein Sci. 2002, 11 (5)
1036-1049) that numerous, indeed perhaps all proteins are capable of forming
potentially disease promoting self-propagating fibrils

There seems with ALS to be a need for interest groups to be made aware of the possible benefit of heparin-like drugs for inhibiting the
formation of the general type of toxic fibril and plaque formation which might play a part in the aetiologies of ALS and Alzheimer¶s
disease (›
c
., U. Lindahl, Haemostasis, 1999, 29, 38-47; D.H. Small c., Ann. New York Acad. Sci., 777, 316-321 Z. Scholefeld c
., J Cell Biol., 2003, 163, 97-107 (›
the possibilities of novel heparin-based therapies for atheroscleosis and cancer (

c›.,
respectively
H. Engelberg, Pharmacol. Rev., 1996, 48 327-352 and, this author, Cancer (New York) 1999, 85, 257-273 (›., also H.P. Sorensen c.,
J. Biol. Chem., 2008, 283, 31920-31932).
Alzheimer¶s disease (AD) is thought to be amenable to therapeutic
intervention by exogenously applied heparin-like polysaccharide drugs [
.
PI 88].
Similarly nvCJD seems to be treatable by pentosan polysulfate [PPS SP54].
This could also indicate that ALS and nvCJD which are also associated with
the formation of toxic fibrils might be treatable by similar mechanisms.
Some doubt has, however, been cast on the primary disease-promoting role of
such fibrils following the results of clinical trials in which the apparent
removal of plaques c   was accomplished by an antibody-based
procedure which apparently did not beneficially affect the progression of AD
in the patients on whom the procedure was tested, however major side effects
were also observed suggesting that much further work is required before the
results could properly be evaluated.
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A phosphorylated form of TDP-43 (a HS-binding protein) (in addition to

misfolded SOD and other proteins) has now been reported to occur as toxic
insoluble inclusions present in the pathologically affected neurons associated
with frontotemporal lobar degeneration (FTLD) (with or without sporadic
ALS) (M. Hasegawa c., Ann. Neurol. 2008, 64 (1) 60-70).

The original notion that altered SOD behaviour could contribute to the
aetiology of ALS was first suggested by D.R Rosen. c., Nature, 1993,
362, 59-62; ›., V.L. Calder c., Neurosci. Lett., 1995, 189 (3) 143-146
who showed that SOD-1 (Cu,Zn SOD) is mutated gene on chromosome 21q
in approximately 15-20 percent of people with familial ALS.
However, rather than the ALD disease process being promoted by diminished
SOD function, as was first thought, later work established that the most likely
effect of mutant SOD in promoting ALS was its pathological misfolding and
subsequent aggregation (which inhibited its expulsion from the cell) c.
It should be noted that such misfolding and functional activity diminution can
be greatly promoted by nitration, especially by tyrosine nitration.
Such   


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3,

 

 

 
 

Nitrosative stress promotes S-nitrosylation (which, with S-glutathionylation)
can deactivate the active site cysteine residues of a key protein folding regulator protein: disulphide isomerase PDI.
Nitration may also disrupt the hydrogen bonding between proteins and water a phenomenon thought to be the
ultimate origin of protein folding and misfolding
(›
 P. Wiggins & G. Steel, Nature Precedings : hdl:10101/npre.2007.1381.1 : Posted 1 Dec 2007) a1 .
Nitrosative stress could tip the balance of PDI activity from the preservation of native state into the promotion of the
aggregation of misfolded PDI proteins in ALS, Parkinson¶s disease and AD ®›
 T. Uehara c., Nature 2006, 441,
513-517} A.K. Walker c., Brain, 2010, 133, 105-116 hinted that PDI mimetics may, by enhancingcPDI-like
activity perhaps also mop up excess nitric oxide be effective anti-ALS therapeutic agents.
Anti-nitrant function is postulated to counter the effects both of tyrosine
nitration as well as aberrant nitrosative perturbation of HS tissue protection
and signalling (perhaps these activities occur in the brain in conjunction with
the DOPA system [DOPA is formed from tyrosine by the addition of an
additional OH group to the aromatic ring to create a catechol derivative]).
Nitric oxide at low concentrations is used as a signalling molecule and at high
concentrations as a cellular immune response (
. to kill invading
pathological organisms). When the latter system gets inappropriately turned
on auto-immune tissue damage can occur.
The amyloid precursor potential plaque -forming proteins seem to be part of
the control system for nitric oxide regulated HS signalling c
-
c

 R. Cappai c. (J. Biol. Chem., 2005, 280 (14) 13913-13920) who
reported that the amyloid precursor protein (APP) of AD and its paralog
APLP2 can critically modulate the Cu/Zn-nitric oxide catalysed de-
polymerisation of HS).
The out-of-control redox metal driven HS dysfunction model could be useful
for a fuller understanding of degenerative disease processes since HS is
involved in wound control and wide-spectrum anti-pathogen activity
(including anti-viral and anti-prion activity ›.
Perturbation of HS signalling  abnormal scission of desulphonated HS is
known to occur in a copper ion and redox-status-determined manner (›., K.
Ding c., J. Biol. Chem., 2002, 277, 33353-33360) as well as, putatively
also in a redox status iron ion determined manner (›., that which was
discussed by D. Grant at http//web.ukonline.co.uk/dgrant/dg2/ where it is also
noted that cytokines which regulate nitric oxide seem to be implicated in
degenerative disease processes which involve abnormal degradation of HS.
Possible roles of trace and ultratrace inorganic elements (
. in commercial
heparin).
It is known that heparin tends to form multi-inorganic element adducts. What is labelled ³sodium‰ or
³lithium‰ heparin can also contain significant amounts of a wide range of inorganic elements. This type
of phenomenon also ion principle also applies to lithium carbonate. The actual active ingredient for some
applications may, at least in part, be due to the presence of such ³impurities‰.

Administration of any commercial heparin irrespective of the labelled designation 

as a lithium,
calcium or sodium salt form could, in principle, provide therapy based on the administration of various
kinds of inorganic ions. 
. the original form of porcine mucosal sodium heparin might be directly
useable as a source of lithium to treat ALS. (A multi-inorganic element analysis of individual heparin
would be required to determine the target inorganic element content).
An Ayurvedic medicine (³Shilagit‰) seems also to be a multi-inorganic element polyanion (it is a
geological fulvic acid multi-element salt apparently derived from similar polyanionic assemblage termed
humic acid salts present in soil organic matter which provide a reservoir of essential inorganic nutrients
required for plant growth). Here also the therapeutic benefits may depend on the co-occurrence of the
organic polymer with a range of inorganic elements.
It is conceivable that the presence in biological fluids of inappropriate amounts of Al3+ (
. aluminium overload)
could also accelerate the misfolded protein aggregation (
. by a general Hofmeister-effect) processes which could
disturb the ability of heparin-like substances to inhibit the role of PSI in promoting this aggregation as well as
augment nitrosative stress.


 cccc#cc".cc

* c
c
cc)c
c cc"c-c
&c
)cccc&cccc!.c5c
c
Lead ions are known to be neurotoxic.
Metal Intoxication May Impact on HS Biochemistry.
®Lead intoxication and HS biochemistry. Pb and Hg intoxication is known to diminish HSPG biosynthesis and
reduce the degree of sulphation. Also, Al3+ and Be2+strongly bind to heparin-like molecules}.

[
H.M. Kurlander c., Metals in spinal cord tissues of patients dying of motor neuron disease, Ann. Neurol.
1979, 6 (1) 21-24, studied photon excited energy dispersive X-ray analysis of spinal ventral horn tissue which
suggested that the degree of Pb intoxication correlated with the duration of the illness].
That anionic polysaccharides from other than animal sources perform detoxification
function c   is suggested by the ability of anionic polysaccharides from algae to behave
in this way allowing for their commercial use for the purification of contaminated water
supplies.
The animal HS proteoglycans systems which contain heparin-like polysaccharide side
chains which (based on c   studies conducted with heparin) are apparently similarly
highly effective multi-inorganic element (including toxic element) sequesters are centrally
involved in wound healing and development including in neurological systems so are
candidates for consideration in the context of unravelling the aetiology of MNDs.
Heavy metal intoxication (especially by lead) has been implicated in the aetiology of ALS and
related diseases.

Mutation in the HFE gene commonly associated with hereditary chromatosis and iron overload
diseases seems to be more prevalent in Alzheimer¶s disease and ALS patients, 

they are
especially susceptible to reactive oxygen , oxidative damage of tissues.
(. reported presentation by J. Connor at the third International Metals and Brain symposium held
in Cape Town during April 2005).

Mercury ions apparently can also enter and damage particular motor neurons.

It has been argued that organic mercury in the form of Thimerosal might contribute to GWS-like
illnesses.

In an epidemiological study of the incidence of multiple sclerosis in North East Scotland it was
proposed by D.I. Shepherd (D.Sc. Thesis University of Aberdeen, 1976) noted that mercury
intoxication should be considered as a prime factor inducing this disease since the first description
of the disease followed the start of widespread dental use of mercury amalgams. An alternative
inorganic element intoxication hypothesis was proposed later based on water, soil and plant tissue
analysis of sites identified by Shepherd. This suggested that that barium intoxication might (also)
have contributed to the environmental intoxication cocktail which may be partly responsible for
inducing this disorder (M. Purdey, Med. Hypoth., 2004, 62 (5) 746-754).

It should be noted that

, action of which is an alternative

generator of HS signalling oligosaccharides, can beneficially regulate the
production of pro-inflammatory cytokines and nitric oxide by 
. T-cells, ›
,
M. Bitan c
 Diabetes Metab. Res. Rev., 2008 24 (5) 413-421;
heparanase de-polymerises HS in a controlled manner, nitric also normally
also de-polymerises HS normally in a highly controlled manner but this non-
enzymic (more primitive) de-polymerisation route seems also to be much
more able to get out of control and thereby to cause inappropriate de-
polymerisation of HS. [The products of HS scission, 
., HS
oligosaccharides, including disaccharides, can rationally be suggested to be
the most likely active agents responsible for the heparanase modifying effect
of T-cell activities].

The cellular immune response to the presence of insoluble plaques ›

including those formed from the amyloid precursor protein of AD which are
unlikely to be normal healthy tissue components, can also, in principle, also
create inappropriate amounts of reactive oxygen and nitric oxide metabolite
levels the presence ofcwhichcis likely to be, at least partly, the origin of
neurodegenerative diseases.

c
The muscle dynein-dynactin ³motor‰ system which apparently normally acts
as a ³garbage disposal‰ system (

for removal of misfolded proteins and
perhaps also toxic metal-ion induced insolubles) could be defective in ALS
patients (›., Y. Goldman c. researches reported in Nanowerk News, July
17, 2008).

 a paper which discusses The interaction of ALS ±related mutant SOD
with the dynein-dynactin microtubule-related retrograde transport
sequestration produces insoluble inclusions in affected neurons (A.L. Strom c
., J. Biol. Chem., 2008, 283, 22795-22805).
The ³taking out the trash‰ from cells is apparently also enabled by heparin-like PPS molecules.

e
Prior to the confirmation of the uniquely effective ability of heparin-like drugs to inhibit prion
disease processes a number of dyestuffs (

Congo red) had been indicated to be of possible
therapeutic value to inhibit this seeded propagation of infective agent (›
 Y Kawasaki c., J.
Viorol., 2007, 81, 12889-12898).
The similar inhibition of solid phase formation by the presence of inhibitors such as organic
dyestuffs is a well-studied phenomenon of long standing interest to colloid chemistry scientists.
A historical textbook ³Colloid & Capillary Chemistry‰ by H. Freundlich (Methuen , London 1926)
discussed at length the role of seeding and inhibition by surface adsorption of ionic inhibitors of
phase change processes including blood coagulation and protein de-naturation.

f
Ideas for Novel Therapeutic Heparin-Related Procedures to Inhibit ALS
Heparan sulphates are heparin-like polysaccharides which are synthesised as proteoglycans in the Golgi
apparatus associated with the endoplasmic reticulum (ER). These polysaccharides apparently exert a
wide-ranging (including water activity regulating) tissue protective effects which include the ability
normalise protein folding which apparently can occur in concert, in an as yet poorly understood way, with
actions of the protein disulphide bond sorter protein disulphide isomerase (PDI) and related proteins.

g
It has also been reported that co-administration of insulin-like growth factor-1
(IGF-1) and low doses of (68% HS, dermatan sulphate mixture of similar
potency to heparin) glycosaminoglycans greatly delays MND in the 
mouse (A. Gorio c., Diabetes, 2002, 81, 194-202).
. also Note 2a

h
In addition to the established need for antioxidant protection there should logically be an additional
need for a possibly separate system of
anti-nitrants to enable the preservation of essential organic molecular
structures in living organisms.
It can be suggested that numerous diseases including ALS could arise from defects in key c  
anti-nitrant protection systems.
Putative roles of the HS system for the regulation of anti-nitrant protection could be of critical
importance to this scenario.
There is urgent need for research efforts to be directed towards finding how
natural antinitrant protection is achieved c   and how this can be corrected for by therapeutic
intervention.
Part of this research should attempt to identify the chemical natural of the most damaging active
nitrogen molecules. It can be rationally suggested that active nitrogen factors other than
peroxynitrite are of pathological significance.
It is currently believed that peroxynitrite generated by the reaction of nitrite with reactive oxygen radicals,
is the key destructive agent and hence the proposed antinitrants are agents which block peroxynitrite
actions and might be effective therapeutic agents (›., Salvemini c., Drug News Perspect, 1998,
11 (4) 204). This idea seems not to have generated the required anti-ALS agents however at the
present time.
This is somewhat surprising since nitric oxide metabolites is known to be centrally implicated in
the pathogenesis of ALS (as well as in stroke, multiple sclerosis, AD, Huntington¶s and
Parkinson¶s diseases).
In ALS, nitrosative stress, in addition to the formation of nitrated tyrosine (also as suggested by the
mouse model of fALS in proteins such as SOD-1,  and h enolase, ATP synthase i chains and
heat shock cognate 71kD protein, ›., F. Casoni c., J Biol. Chem., 205 280 (10) 16295-16304)
also leads to S-nitrosylation and S-glutathionylation of the active site cysteine residues in PDI.
This mechanism could to be how nitrosative stress tips the balance of PDI activity from the
preservation of native state into the promotion of the aggregation of misfolded PDI proteins in
ALS, Parkinson¶s disease and AD ®›
 T. Uehara c., Nature 2006, 441, 513-517}.
A recent report (A.K. Walker c., Brain 2010, 133, 105-116) suggested
that the process of mutated SOD aggregation and therefore ALS can be
inhibited by


V
$,
suggesting that modification of PDI activity so as to inhibit ALS might be
achieved by the exogenous application of PDI active site mimetics such as
V'

 

V
V

!
(such mimetics may 

create more PDI-like activity or even, it can further
be suggested, by mopping up excess nitric oxide cancact as anti-nitrants).
c

Nitrosative stress is, in general, characterized by the nitration of tyrosine amino acid moieties in proteins
but perhaps also the free amino acid. A less-well known, but perhaps critically important effect of
nitrosative stress could, however, be the inappropriate scission of HS by nitric oxide metabolites. This is
somewhat analogous to the pathological de-polymerization of HS by hydroxyl radicals (
. in
experimental nephrotic syndrome as reported by
C.J.I. Raats c., J. Biol. Chem., 1997, 272, 26734-26741).
Nitrotyrosine accumulation occurs in the aged brain and in brains of patients with AD, multiple sclerosis,
ALS, CO toxicity stroke, and head trauma; ›., G. Deng c., J. Am. Soc. Nephrol., 2001, 12, 1892-
1899, who also found that increased tyrosine nitration occurred in the brain in chronic renal insufficiency
studied in a rat model but this could be reversed by angiotensin-converting enzyme (ACE) inhibition.
(ACE inhibition is also known to preserve HS
PG in the glomerular basement membrane in rats with established adramycin-induced nephropathy (
.,
F.H. Wapstra et al., Exptl. Neprol., 2001, 9(1) 21-2 ). Recently it has been reported (M. Kawajiri c.,
Acta Neurol. Scand., 2009 (5) 341-344) that angiotensin II levels in the CSF of ALS patients is
significantly reduced and that this reduction is correlated with the severity of the disease. These findings
can support a novel stratagem, involving modulation of HS and ACE, to allow a rational treatment of
ALS.
Is blood pressure and electrolyte balance centrally involved here? [It should be noted that the efficiency of
the HS system has a large influence on these functions].
It has been indicated (Y. Iwasaki c., Neurol. Res. 2002, 24 (5) 468-472) that olmesartan a novel
ATR1R agonist protects spinal motor neurons in the rat.
While the degree of protein nitration consequent on renal insufficiency ›. might have been expected to
have been associated with the aetiology of ALS and other neurological diseases, evidence for protein
nitration or oxidative damage (
. as hydroxyl radical effects which show up as malonaldehyde) seemed
to be lacking in ALS, a circumstance which could engender doubt about the relevance of peroxynitrite
damage in this disease.
Perhaps this is because peroxynitrite is the wrong target;
[›
c
g, M.G. Espey c
, PNAS USA, 2002 99 (6) 3481-3486 and S. Pfeiffer & B. Mayer, J.
Biol. Chem., 1998, 273 (42) 27280-27285];
(›.also , W. Robberecht, J. Neurol., 2000, 247 (suppl-1)-7/1-6 .
Other workers, (L.J. Bruijn c., PNAS, USA, 1997, 94, 760-766) however, found evidence for the
occurrence of a 2-3 fold increase in nitrotyrosine formation in ALS which coincides with the earliest
pathological abnormalities and remains a feature of the spinal cord environment of ALS patients
throughout the progression of the disease. The possibility exists that unconventional tyrosine nitration as
well as the nitrosative scission of HS may underpin the aetiology of ALS.

Active nitrogen tissue damage in ALS is putatively mediated, in addition to

protein nitration (
. as discussed by F. Casoni c., J. Biol. Chem. 2005,
280 (10) 16295-16304) probably also  the formation of nitrated tyrosine
monomer. (
the additionally hydroxylated tyrosine, with an additional
adjacent OH group to the 3-OH group of tyrosine is the molecule DOPA
which when decarboxylated to dopamine apparently might act synergistically
with cysteine in neuromelanin, a substance which is thought to act as a
multiple target protectant antioxidant ›., (it could be a key µantinitrant¶) and
a sequester of 
. chlorinated pesticide pro-inflammatory agents such as
HCB. Neuromelanin seems to be an example of the wider system of
cysteinyl-tyrosine couplings in proteins which apparently confer antioxidant
(and putatively antinitrant) protection (›., D. Grant c., Med. Hypoth.,
1989, 28, 245-253). Defects in such systems could underlie the aetiologies of
numerous degenerative diseases.

Excessive Nitrosative Stress Putatively Perturbs HS Tissue Protection

It should be noted that nitrosative stress (especially coupled with copper
dyshomoestasis) is also capable of inappropriately causing nitrosative
scission of HS, a process which generates oligosaccharides which can
penetrate into cells. Nitrosative stress can also cause altered sulphation of
HS expecially a reduction of N-sulfation (  the occurrence of non-enzymic
scission of the S-N bonds). Under inappropriately prolonged nitrosative stress
episodes it is therefore likely that inappropriate scission of HS may occur.
The normal HS signalling system seems to include an important feedback
mechanism which utilises the ability of nitric oxide metabolites to cleave HS
at pre-designated sites. This system is likely to become unbalanced under
conditions of excessive nitric oxide formation.
This mechanism may be part of the complex interplay of dysfunctions
involving both nitric oxide and metal ion biochemistry which underlie the
aetiologies of ALS, other neurological diseases and autoimmune and
degenerative diseases in general (›., D. Grant,
http//web.ukonline.co.uk/dg2/)

A possible instance of the above scenario is that the kinds of HS molecules

which have been reported (I.B. Bruinsma c., Acta Neuropathol., 2009,
PMID 19636575) to be associated with the kinds of insoluble fibrils which
are present in AD brains, have an altered microstructure, having
c ccccc›
ccc .c
›cc›c c 
c c  c›

This suggests that defects of N-sulfation, 
. the occurrence of postsynthetic
desulfation which can occur non-enzymatically by the catalytic action of nitric
oxide metabolites in the presence of unliganded iron ions, H+ ions (partly
unpublished results Aberdeen University) or following N-chlorination in the
presence of myeloperoxidase (›
M.D. Rees and M.J. Davies, J. Amer.
Chem. Soc., 2006, 128 (9) 3085-3097) may counter the normal ability of fully
N-sulphated HS to inhibit the formation of various kinds of pathological
plaques. This mechanism may contribute to the process by which unliganded
iron ions and oxidative stress including where chlorination can occur, can
promote AD and other neurodegenerative diseases as well, perhaps, as some
kinds of cancer (›. D. Grant c., Med. Hypoth., 1992, 38, 49-55).
Chlorination of HS is also a possible outcome of intoxication and lifetime
increment within human tissues of man-made chlorinated organic molecules
(
. those formerly used as insecticides which are now banned by
international agreement which, however continued to be used in developing
countries and transported intercontinentally in the stratosphere as well as
synthesised
   in large quantities in municipal and medical incinerators
(›. D. Grant, internet document accessed  Google search term ³Amey Bishop hexachlorobenzene‰
which more specifically discusses how the dioxin-like substance hexachlorobenzene which is commonly
produced during combustion processes and following its bio-concentration up the food chain promotes
numerous diseases in humans).It can be also putatively suggested that the intoxication by such substances
may be implicated in the etiology of ALS as well as in the range of other diseases discussed in this
document).

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(Anionic) polysaccharides, polyihydroxybutyrates and inorganic polyphosphates are known
more generally to be associated with all types of cell surfaces throughout biota and seem to occur
together with HS polysaccharides at animal membranes (the presence and role of HS with
mitochondria is currently not well established however). One possible function of such polyanionic
substances as well as a principal function of being centrally involved in energy metabolism, is for
creating ion channels (›. for polyphosphate-related systems in mitochondrial membranes as
reported by A.Y. Abramov c., PNAS USA, 2007, 45, 18091-18096) could also be of use for
the gathering of essential nutrients as well as for the provision of a protective barrier against
various insults including the entry of toxic elements such as mercury and cadmium into organelles
as well as into whole cells).
Cell surface anionic polysaccharides and polyihydroxybutyrate and related polyesters could also
function as heavy metal intoxicant filters, and defects in such protection might promote those
diseases like ALS which are, at least in part, thought to be promoted by toxic metal ions.

Activation of a bivalent ion sensitive ion channel under oxidative stress including that which is
glutamate induced is also, however, thought to be how cellular apoptosis occurs in MNDs.
Experimental evidence has recently been obtained that mitochondrial cell membrane
polyihydroxybutyrate could also be directly involved in this process since these polymers have
been found to cause a calcium ion induced cell suicide (apoptosis) trigger in mitochondria. (It
should be noted that mitochondria are believed to be descended from early eukaryote incorporation
of symbiotic bacteria as is still the case with nitrogen fixation plants; such bacteria always contain
cell surface polyihydroxybutyrate where they analogous to cell surface polysaccharides which can
rise to give glucose, can serve as reservoirs of hydroxybutryate which can enter the energy supply
cycle for production of ATP).

* Amyotrophic lateral sclerosis (ALS)

Common MNDs include ALS, progressive bulbar palsy, primary lateral
sclerosis and progressive muscular atrophy. Other MNDs include the many
inherited forms of spinal muscular atrophy and post-polio syndrome.

*This fatal neurodegenerative disease is the most common form of MND and
is characterised by spinal and cortical motor neuron degeneration.


This document is not meant to substitute for medical advice and affected
individuals should always consult medical practitioners to obtain such advice.
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