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Convergence
Olly
Posted: Tuesday, June 12, 2012 7:20:23 AM

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Convergence
A reasonable definition would be:
'the degree or point at which lines, objects, etc., converge.'

Why is it so important in ALS?
Well though many things may possible start off ALS there should be a stage where the causes and effect of genetic and sporadic ALS converge to cause destruction of the neurons.
That sounds logical and if true would go a long way to helping with developing a downstream therapeutic for ALS, if we can find the convergent point.

But is it true and can we find any level of convergence or is it just a mirage?

For instance there are many ways I can stop an electrical wire from working; by cutting, using, acid, electrical overload, fire etc and could the same be true in ALS?

I am ignoring inflammation which seems to be common in all cases of ALS and is a reaction to system dysfunction and injury.

I want to start this thread to see if we can identify common pathways which arise in all the different types or subsets of ALS, other than inflammation and will add to it when I can.

Unlike Captain Oats in the Antarctic I may be gone some time but will be back to post in this thread.

Please feel free to post if you find any convergence or commonality in the pathological processes of ALS.


Into the heart, an air that kills, from yon far country blows.
What are those blue remembered hills, what sphires what farms are those.
That is the land of lost content,I see it shining plain,
The happy highways where I went and cannot come again
DeeBee
Posted: Tuesday, June 12, 2012 8:30:31 AM

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My suspicion is that MND is a 'genetic weakness' because most of us are too closely related and therefore lack hybrid vigor. The faulty genes are probably from the distant past and 'well up' when the time is right. The symptoms of sporadic and familial ALS are more or less the same so the root must also be very similar?

( It would be a shame if 'enthusiasts' do not take up one of the many 'collaborative research' offers being advertised by labs and universities at the moment ).
Olly
Posted: Tuesday, June 12, 2012 9:35:48 AM

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The following has been cut and pasted from other threads but may show convergant processes in ALS concerning VEGF, ANG by upregulating Bcl-2, which inhibits caspase-3.

Vascular endothelial cell growth factor (VEGF) requires ANG rRNA transcription to function correctly to protect neurons. Bcl-2 and Nf-κb pathways are also involved, so are all these pathways common to most ALS cases?


Molecular biology of amyotrophic lateral sclerosis: insights from genetics

Complex genetics in ALS.
There have been few studies of genetic variants that modify ALS susceptibility or phenotype.

In DNA sets from Sweden, Belgium and Birmingham (UK)44, but not London (UK)44, Utrecht (The Netherlands)45 or Boston (USA) (R.H.B.,unpublished data), promoter polymorphisms that reduce the expression of vascular endothelial growth factor (VEGF) are associated with an increased risk of disease.

Variants predicted to reduce the activity of another vascularizing factor, angiogenin (ANG), increase ALS risk selectively in Irish and Scottish DNA sets46.

These observations support the view that vascularization and, indirectly, blood supply and cellular oxygen pressure are important determinants of motor neuron viability,
or that VEGF and ANG exert direct neurotrophic influences on motor neurons.

http://www.researchals.org/uploaded_files/pasinelli_nns_rev_0906_844ub6.pdf

ANG moves to the nuclear in endothelial cells where it binds a to a region of rDNA which then stimulates rRNA transcription. Vascular endothelial cell growth factor (VEGF) requires ANG rRNA transcription to function correctly to protect neurons. VEGF protects motor neurons from ischemic injury, death and if disrupted motor neurons degenerate.
...............................
Angiogenin is found in a variety of cells throughout the body and circulates in the bloodstream. When angiogenin binds to receptors on the surface of endothelial cells, it triggers a series of reactions that brings angiogenin inside the cell.

Once inside endothelial cells, angiogenin moves to the nucleus where it stimulates the production of ribosomal RNA (rRNA), a chemical cousin of DNA. Ribosomal RNA is required for assembling protein building blocks (amino acids) into functioning proteins.

Angiogenin stimulates the production of additional ribosomal RNA when there is an increased demand for proteins, such as for the growth and division of endothelial cells. Angiogenin may also be involved in other steps of angiogenesis, such as helping to break down the tissue that surrounds existing blood vessels to allow room for the growth of new blood vessels
Angiogenin is a liver-derived component of normal serum the concentration of which can increase in various disease states and its expression is regulated in vivo in a manner that is characteristic of acute phase proteins (Olson et al, 1998).

In the current work, Li and Hu lay out the biochemical pathway linking angiogenin and apoptosis inducing factor (AIF). AIF is normally located in mitochondria, but when it moves to the nucleus—as it does in ALS model mice (Oh et al., 2006)—it chews up DNA, contributing to apoptosis.
The following indicates that angiogenin stops AIF moving to the nucleus by up regulating Bcl-2 which inhibits caspase-3 ......Therefore giving an explanation of the death or destructive pathway.

Critically is this common in most ALS cases albeit by slightly different but related pathways and the cause of neuron death?

(In addition to angiogenin and AIF, the players include the apoptosis dampener Bcl-2 and the pro-apoptosis proteins caspase-3 and serum polymerase-1 (PARP-1).

The researchers delineate a pathway in which apoptosis normally proceeds from activation of caspase-3, to cleavage of PARP-1, to nuclear translocation of AIF.

Angiogenin alters the situation by upregulating Bcl-2, which inhibits caspase-3 and thus the rest of the downstream pathway. Much of this pathway was already known, noted Piera Pasinelli of Thomas Jefferson University in Philadelphia, Pennsylvania, but “they really teased out the mechanism by which angiogenin can be anti-apoptotic…in a Bcl-2-dependent manner,” she said.

To study apoptosis, the researchers starved P19 cells of serum. This treatment normally causes PARP-1 cleavage within a few hours and nuclear translocation of AIF by 24. But with angiogenin in the culture media, PARP-1 stayed whole and AIF remained mitochondrial, confirming that angiogenin blocks apoptosis via this pathway. In a previous study, the authors confirmed that angiogenin also prevents activation of caspase-3 (Li et al., 2010).

The researchers already knew Bcl-2 was upregulated by angiogenin (Li et al., 2010), and suspected it would be crucial to angiogenin’s effects. If so, then removing Bcl-2 should allow apoptosis to proceed unhindered, even in the presence of angiogenin. Li used RNA interference to knock down Bcl-2. In the knocked down, serum-starved cell cultures, angiogenin was less effective at preventing caspase-3 activation, PARP-1 cleavage and AIF translocation.)
Notice in the above the researchers said, in the last paragraph, that angiogenin was less effective which implies that it did not totally stop caspase-3 activation so is there another factor at work?

Angiogenin upregulates anti-apoptotic genes, including Bag1, Bcl-2, Hells, Nf-κb and Ripk1, and downregulates pro-apoptotic genes, such as Bak1, Tnf, Tnfr, Traf1 and Trp63.

Knockdown of Bcl-2 largely abolishes the anti-apoptotic activity of angiogenin, whereas the inhibition of Nf-κb activity results in a partial, but significant, inhibition of the protective activity of angiogenin.

Thus, angiogenin prevents stress-induced cell death through both the Bcl-2 and Nf-κb pathways.


From the above it looks like upregulating Bcl-2 and Nf-kb through some other method may also help us besides increasing angiogenin levels?


Into the heart, an air that kills, from yon far country blows.
What are those blue remembered hills, what sphires what farms are those.
That is the land of lost content,I see it shining plain,
The happy highways where I went and cannot come again
lochnerd
Posted: Wednesday, June 13, 2012 4:01:38 PM
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More biochemistry complexity. I hope you find this useful, and perhaps relevant to your thoughtful threads.


Protein Interaction Profiling of the p97 Adaptor UBXD1 Points to a Role for the Complex in Modulating ERGIC-53 Trafficking*

Dale S. Haines‡§,**,
J. Eugene Lee¶,
Stephen L. Beauparlant‡,
Dane B. Kyle‡,
Willem den Besten¶,
Michael J. Sweredoski‖,
Robert L. J. Graham‖,
Sonja Hess‖ and
Raymond J. Deshaies¶

+ Author Affiliations

From the ‡Fels Institute for Cancer Research and Molecular Biology and
§Department of Biochemistry, Temple University School of Medicine, 3307 North Broad Street, Philadelphia, 19104 Pennsylvania;
¶Howard Hughes Medical Institute and
‖Proteome Exploration Laboratory; Beckman Institute, California Institute of Technology, 1200 E California Blvd. Pasadena, 91125 California

↵** To whom correspondence should be addressed: Fels Institute for Cancer Research and Molecular Biology Temple, University School of Medicine, 3307 North Broad Street, Philadelphia PA 19104. E-mail: dhaines@temple.edu.

Abstract

UBXD1 is a member of the poorly understood subfamily of p97 adaptors that do not harbor a ubiquitin association domain or bind ubiquitin-modified proteins. Of clinical importance, p97 mutants found in familial neurodegenerative conditions Inclusion Body Myopathy Paget's disease of the bone and/or Frontotemporal Dementia and Amyotrophic Lateral Sclerosis are defective at interacting with UBXD1, indicating that functions regulated by a p97-UBXD1 complex are altered in these diseases. We have performed liquid chromatography-mass spectrometric analysis of UBXD1-interacting proteins to identify pathways in which UBXD1 functions. UBXD1 displays prominent association with ERGIC-53, a hexameric type I integral membrane protein that functions in protein trafficking. The UBXD1-ERGIC-53 interaction requires the N-terminal 10 residues of UBXD1 and the C-terminal cytoplasmic 12 amino acid tail of ERGIC-53. Use of p97 and E1 enzyme inhibitors indicate that complex formation between UBXD1 and ERGIC-53 requires the ATPase activity of p97, but not ubiquitin modification. We also performed SILAC-based quantitative proteomic profiling to identify ERGIC-53 interacting proteins. This analysis identified known (e.g. COPI subunits) and novel (Rab3GAP1/2 complex involved in the fusion of vesicles at the cell membrane) interactions that are also mediated through the C terminus of the protein. Immunoprecipitation and Western blotting analysis confirmed the proteomic interaction data and it also revealed that an UBXD1-Rab3GAP association requires the ERGIC-53 binding domain of UBXD1. Localization studies indicate that UBXD1 modules the sub-cellular trafficking of ERGIC-53, including promoting movement to the cell membrane. We propose that p97-UBXD1 modulates the trafficking of ERGIC-53-containing vesicles by controlling the interaction of transport factors with the cytoplasmic tail of ERGIC-53.
Footnotes

↵* DSH acknowledges support from the Fels Institute for Cancer Research and Molecular Biology and the Temple University Faculty Senate Seed funds. JEL was supported by the Ruth L. Kirschstein NRSA fellowship CA138126 whereas WBD was supported by the Ruth L. Kirschstein NRSA fellowship GM088975. The Proteome Exploration Lab was supported by the Beckman Institute at Caltech and an award from the Gordon and Betty Moore Foundation. RJD is an Investigator of the Howard Hughes Medical Institute and this work was supported in part by HHMI.

↵Graphic This article contains supplemental Tables S1 to S5.

Received December 12, 2011.

© 2012 by The American Society for Biochemistry and Molecular Biology, Inc.
is relevant. I get many articles sent to me and just want to share. Mark
DeeBee
Posted: Thursday, June 14, 2012 5:40:33 AM

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DeeBee wrote:
My suspicion is that MND is a 'genetic weakness' because most of us are too closely related and therefore lack hybrid vigor. The faulty genes are probably from the distant past and 'well up' when the time is right. The symptoms of sporadic and familial ALS are more or less the same so the root must also be very similar?

( It would be a shame if 'enthusiasts' do not take up one of the many 'collaborative research' offers being advertised by labs and universities at the moment ).


i.e.

http://www.mndassociation.org/research/MND+research+and+you/Get+involved+in+research

(copy&paste)
David Hicks
Posted: Sunday, June 17, 2012 2:58:38 AM

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ALS/MND is a multifaceted disease with multiple causes. Many people have proposed this many times, while others persue the idea that there must be one single cause. My personal research has led me to believe that there are multiple causes for ALS and all of these causes are linked by one "COMMON EVENT" (or a convergence point). Now don’t go getting excited that a common event has finally been discovered. Without proof, no one is going to believe it unless it is backed up with maybe a hundred examples to show how the common event is the start of ALS. I have been working on compiling such a list over the last two and a half years and I am still working on it. This involves many new discoveries and I expect to write over 100 pages of detail, so it is not going to be fully explained at this time in this post. You should notice that the common event not only helps explain different phenotypes of ALS, it is also the explanation of many other diseases.

My proposal on “the common event” and “the chronic failure of cellular regeneration theory” is given below in the hope that ALS TDI may at least start to seriously look at the possibility of the COMMON EVENT. I would like to use this post to ask ALS TDI if they could list say three different things about sporadic ALS that at present cannot be explained or has conflicting outcomes in research. I will take it as a challenge and try to explain one or all three while showing how they would connect back to the common event.

If you are just looking at ALS, the common event guides you to the realization that all ALS is caused by a chronic failure of the first growth phase of cellular regeneration (G1/S), from progressing to the second growth phase (G2/M).

CHRONIC FAILURE OF CELLULAR REGENERATION! THE COMMON EVENT.

Chronic failure of cellular regeneration is just five words; but the study of cellular regeneration can lead to the understanding that CHRONIC FIRST PHASE cellular regeneration failure (G1/S) can lead to dozens of diseases including cancer, diabetes and neurological diseases. It can also exacerbate other diseases like malaria and HIV. The solution to understanding the differentiation of different diseases is reliant on knowing which type of cell is in need of regeneration. Every different somatic cell requires a different regeneration “recipe”. The difference in the cell type, and therefore the "recipe" can cause different diseases. Chronic regeneration failure leads to cellular degeneration. “Regeneration failure” is presently mistakenly defined as degeneration, but that definition misleads researchers into looking at degeneration instead of cellular regeneration failure.

HYPOTHESIS NO.1 OR “CHRONIC FAILURE OF CELLULAR REGENERATION THEORY”.

Many diseases including cancer, diabetes and neurological diseases, and others that are currently thought of as inflammatory-like conditions are the result of A CHRONIC FAILURE OF CELLULAR REGENERATION. The chronic failure acts as a cell-specific precursor causal agent.

The above is a cell-specific vulnerability rather than a propensity to cause a specific disease. This covers the actions of chronic cellular regeneration in both familial and sporadic conditions. Every altered action can be explained and traced back through their pathways to the start or failure of this repair mechanism. This hypothesis covers all causes and all diseases mentioned above. It includes physical and non-physical chronic cellular insult as well as the failure of cellular repair due to familial defects. Diseases such as type 2 diabetes, some cardiovascular disease and stroke can eventuate by a singular action of a failure of chronic cellular regeneration. Other diseases such as cancer and AIDS require a concomitant or 2 hit cause. Pathogenic diseases can be compromised by a chronic regeneration failure of the repair mechanism that leads to a down regulated immune system.

Every changed setting required for the start of a quick cellular injury repair will produce the exact requirement for cancer growth. This in itself will not result in cancer, but; for specific cell types; if you introduce a chronic insult, familial mutation, virus, mutagen etc., it can result in cancer if these settings become chronic or near permanent over a long time frame of years. In adult conditions it will give the body time to produce a mutated cell. DNA errors are produced in about one in each 10-100 billion chromosome base pairs. These mutations will escape the immune system in adult/sporadic conditions when the early regeneration settings down regulate apoptosis, activates angiogenesis and promotes motility and cell growth (for G1 repair). A long time period will also allow further mutations of an already mutated cell.




David Hicks.
Olly
Posted: Wednesday, June 20, 2012 12:23:59 PM

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Looking at VEGF again and convergant processes in ALS ..

Downregulation of genes with a function in axon outgrowth and synapse formation in motor neurones of the VEGFδ/δ mouse model of amyotrophic lateral sclerosis

Abstract

Background

Vascular endothelial growth factor (VEGF) is an endothelial cell mitogen that stimulates vasculogenesis.

It has also been shown to act as a neurotrophic factor in vitro and in vivo.

Deletion of the hypoxia response element of the promoter region of the gene encoding VEGF in mice causes a reduction in neural VEGF expression, and results in adult-onset motor neurone degeneration that resembles amyotrophic lateral sclerosis (ALS).

Investigating the molecular pathways to neurodegeneration in the VEGFδ/δ mouse model of ALS may improve understanding of the mechanisms of motor neurone death in the human disease.

Conclusions

Reduction in expression of VEGF, through deletion of a regulatory promoter region of the gene, results in adult-onset motor neurone degeneration that resembles human ALS.

We have presented evidence here that this phenotype is accompanied by reduction in expression, from symptom onset, of the cholesterol synthesis pathway, and genes involved in nervous system development, including axonogenesis, synapse formation, growth factor signalling pathways, cell adhesion and microtubule-based processes.

These findings raise the possibility that VEGF is required for the maintenance of distal neuronal processes in the adult animal, perhaps through promotion of remodelling of distal processes and synapses in the face of the demands of neuronal plasticity.

A reduction in VEGF expression in VEGFδ/δ mice may lead to failure of the maintenance of neuronal circuitry, causing axonal retraction and cell death.


http://www.biomedcentral.com/1471-2164/11/203




Into the heart, an air that kills, from yon far country blows.
What are those blue remembered hills, what sphires what farms are those.
That is the land of lost content,I see it shining plain,
The happy highways where I went and cannot come again
Nemesis
Posted: Wednesday, June 20, 2012 5:29:33 PM

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Don't want to be too "von oben", but I can't help to notice that ALS has already coverged on vascular rarefaction.

The issue at hand is not to re-converge, but rather how to address the actual problem.

So far, I only see Ichisan trying, which is somewhat strange.

How hard can it be, focus is on;

A) Address acute neuroinflammation.

B) Address revascularization.
.
Let's focus at that.


Don't just ask what scientists can do to speed up the solution for ALS or when they will do it, instead ask yourself what you can do right now to solve ALS asap.
Olly
Posted: Wednesday, June 20, 2012 6:07:19 PM

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No that is not what this thread is about.

If you re read the opening posting this thread states many things may possible start off ALS there should be a stage where the causes and effect of genetic and sporadic ALS converge to cause destruction of the neurons.

You may have bought into vascular rarefaction as a common factor but that doesn't mean it's true, especially given the different subtypes of ALS.

As you state:

'How hard can it be, focus is on;

A) Address acute neuroinflammation.

B) Address revascularization;

It must be very difficult given that you have not solved that problem but I leave that to you and another thread.

By trying to push any research, by others, into what you regard or think as what is required closes down any other approaches to dealing with ALS and is not in the spirit of this site.


Into the heart, an air that kills, from yon far country blows.
What are those blue remembered hills, what sphires what farms are those.
That is the land of lost content,I see it shining plain,
The happy highways where I went and cannot come again
ImInAwe
Posted: Wednesday, June 20, 2012 8:25:11 PM
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Olly wrote:


By trying to push any research, by others, into what you regard or think as what is required closes down any other approaches to dealing with ALS and is not in the spirit of this site.


Actually, I would disagree with this comment. I've read most of the microvascular ischemic/hypoxic threads that Nemesis has written, and I've not ever interpreted his hypotheses and research as being pushed on to anyone. Not ever.

Unfortunately, he comes across harsh and unkind at times to certain (awesome) members of this forum.

But his bottom line should not be missed. Inflammation and revascularization. This does not mean that everyone will or even should buy into this idea. It means that among many, many, many potential therapies and mechanisms, it is only one. And it is one that is shared (so it seems) by very, very, few people.

There are 3 reasons why I still come back to read this forum. One, is the curiosity of whether this hypothesis is shared by ALS researchers, and if so, are there are any treatments being pursued to that regard?

Regarding these ideas (microvascular ischemia/hypoxia/revascularization), Nemesis is pretty much stand alone. The only other person that appears to buy into these ideas is Louis. (I've bought into these ideas for the past 17 years, but certainly not with the understanding and scientific precision as described on this site. My opinion doesn't count anyway since I'm not a PALS and no longer a CALS).

When Nemesis expresses his surprise that there are no others (at least that post here, or that post on PLM) doing what Louis' wife is doing: ALCAR, Taurine, Magnesium, hormone therapy, L-arginine, L-lysine, Phosphatidyl choline, Niacinamide, oxygen therapy (hopefully supported and monitored by a primary care physician), and the avoidance of vasoconstrictors, I have to concur with Nemesis. I am also surprised by this. I don't see this particular combination of amino acids/nutrients on PLM either.

That doesn't mean that this particular cocktail will help anyone other than Louis' wife. It doesn't even mean that the (wonderful) stability that Louis' wife is experiencing is due to what he is administering to her. For all we know, it may be a “coincidental” 5 month plateau in her disease progress that has nothing to do with the present treatment regime that Louis has her on. But in a disease where it must feel like your back is up against a wall at times, I am surprised that this combination is only being done by Louis (that we know of).

Many people here are always trying new things. How wonderful for Ben and ENV that Nuedexta seems to be helping them. May the medicine continue to help them.

The second reason I read this forum, is that I learn a lot. In fact, I would have to thank Nemesis for his blurb on angiotensin II inhibitors and his comparison between diabetic retinopathy and the similiarity between it and the hypothetical idea of microvascular ischemia in ALS. I saw a 32 year old woman with severe proliferative diabetic retinopathy. She was having severe retinal bleeds every 1 – 2 weeks with complete vision loss followed by slow recovery for the past 2 years. Three opthamologists, an ocular surgeon, laser treatments, bilateral vitrectomies, and multiple Avastin injections later, her condition had not improved and continued to deteriorate. After reading Nemesis' ideas, I switched her ACE inhibitor to Cozaar, and used therapies that improved microcirculation like very high doses of proanthocyanidins, flavonoids, etc., etc., etc. Since she started this, she has had no retinal bleeds for 4 months. Sure, it could just be coincidence. It could also be coincidental that her mother is a diabetic and her maternal grandmother had died of ALS.

All anybody can ever do is experiment and share. That appears to be the spirit of this site and people can take or leave what they choose.

My fervent hope, is that IF there is indeed something to this hypothetical microvascular ulcerative ischemic syndrome hypothesis, that the brilliant minds of ALS reasearch will explore its possibilities.
Olly
Posted: Wednesday, June 20, 2012 9:03:55 PM

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I would disagree with you ImInAwe with your posting.

This thread was about convergence or commonality in the pathological processes of ALS.

Nem indicate that we should concentrate on addressing acute neuroinflammation and address revascularization and concentrate on those two.

There are plenty of threads on those subjects so why tell me basically to just concentrate on them unless it's pushing his own agenda?

I stated in the first post that I was ignoring inflammation which seems to be common in all cases of ALS and is a reaction to system dysfunction and injury.

I want to start this thread to see if we can identify other common pathways which arise in all the different types or subsets of ALS, other than inflammation and will add to it when I can.

You stated his bottom line should not be missed, inflammation and revascularization, I agree but this thread is not about inflammation and not solely on vasculation.


Into the heart, an air that kills, from yon far country blows.
What are those blue remembered hills, what sphires what farms are those.
That is the land of lost content,I see it shining plain,
The happy highways where I went and cannot come again
Olly
Posted: Wednesday, June 20, 2012 9:51:48 PM

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Bringing SOD 1 into the fold

The study of neurodegenerative diseases, including ALS, has benefited greatly from genetic models that are based on inherited mutations in disease-associated genes. However, for several disorders, including Alzheimer’s disease, Parkinson’s disease and ALS, familial syndromes account for only a minority of the total disease burden13.

Furthermore, SOD1 purified from tissue samples of individuals with sALS, but not SOD1 isolated from control samples, inhibited axonal transport. The presence of C4F6-reactive SOD1 in tissue samples of individuals with ALS and the ability of this conformer to suppress axonal transport suggest that abnormal conformations of SOD1 could indeed mediate motor neuron toxicity in sALS.
Accumulating evidence points toward abnormalities in axonal transport as a final common pathway in motor neuron disease. Deficits in axonal transport might occur through three independent mechanisms
(Fig. 1). Initially, axonal transport could be prevented by improper transcription of essential genes, abnormal processing of the gene transcripts or translational inhibition.

This mechanism likely underlies motor neuron disease as a result of mutations in nucleic acid–binding proteins, such as TDP-43, FUS/TLS, SETX and SMN7.

The intracellular trafficking of microtubules and endosomes is also essential for proper axonal transport and mutations in the genes encoding proteins important for this process (Alsin, Atlastin, VAPB and TBCE) likewise cause motor neuron degeneration8,9.

Mutations in the genes encoding kinesins (KIF1B, KIF5A and KIF21A) and the dynactin complex (DCTN1) directly suppress axonal transport by disrupting motor protein function10. Bosco et al.3 found that mSOD1, found in fALS due to SOD1 mutations,^ and misfolded SOD1, created by oxidation of SOD1, specifically inhibit anterograde axonal transport through disproportionate activation of p38 kinase.

Suppression of axonal transport is lethal in motor neurons and p38 kinase activity is strongly associated with motor neuron degeneration in animal models of ALS11, confirming the potential of misfolded SOD1 to cause motor neuron disease through the same pathway.

Indeed, small molecule inhibitors of p38 kinase show promise in animal models of ALS12, highlighting this mechanism as a major target for drug development in humans.

Are the genetic models that we commonly employ accurate representations of the pathogenesis that occurs in sporadic disease and can we use these models to identify new therapeutics?
The clinical and genetic heterogeneity of motor neuron disease implies that ALS may
not be a single disorder2.

Instead, what we call ALS may represent a collection of separate diseases that are characterized pathologically by motor neuron loss and clinically by weakness.

Nevertheless, the results of Bosco et al.3 suggest that the pathogenesis of fALS linked to SOD1 mutations is, in fact, closely linked to that of sALS.

This is reassuring evidence in support of the relevance of genetic ALS models to sporadic disease.

Inhibition of axon transport may be the final common pathway in motor neuron disease, one that is susceptible to genetic and environmental insults (such as oxidation), perhaps representing a pathogenic mechanism that is common to both the familial and sporadic forms of ALS (Fig. 1).


http://www.projectals.org/pdfs/Barmada_FinkbeinerNV.pdf

Into the heart, an air that kills, from yon far country blows.
What are those blue remembered hills, what sphires what farms are those.
That is the land of lost content,I see it shining plain,
The happy highways where I went and cannot come again
Nemesis
Posted: Thursday, June 21, 2012 8:07:45 AM

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Posts: 3,091

David calls it “the chronic failure of cellular regeneration” I would call it pathologically accelerated replicative senescence. Whatever it should be called, here is a paper related to the subject that David might be interested to read:


FEBS Lett. 2007 Jun 12;581(14):2727-32. Epub 2007 May 21.
The IGF-I splice variant MGF increases progenitor cells in ALS, dystrophic, and normal muscle.
Ates K, Yang SY, Orrell RW, Sinanan AC, Simons P, Solomon A, Beech S, Goldspink G, Lewis MP.

Abstract
The effects of muscle splice variants of insulin-like growth factor I (IGF-I) on proliferation and differentiation were studied in human primary muscle cell cultures from healthy subjects as well as from muscular dystrophy and ALS patients. Although the initial numbers of mononucleated progenitor cells expressing desmin were lower in diseased muscle, the E domain peptide of IGF-IEc (MGF) significantly increased the numbers of progenitor cells in healthy and diseased muscle. IGF-I significantly enhances myogenic differentiation whereas MGF E peptide blocks this pathway, resulting in an increased progenitor (stem) cell pool and thus potentially facilitating repair and maintenance of this postmitotic tissue.

PubMed

Don't just ask what scientists can do to speed up the solution for ALS or when they will do it, instead ask yourself what you can do right now to solve ALS asap.
Olly
Posted: Thursday, June 21, 2012 8:26:28 AM

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Here is a very good and well laid out paper on convergence:

Altered Gene Expression, Mitochondrial Damage and Oxidative Stress: Converging Routes in Motor Neuron Degeneration

Relevant chapters are:

1. Introduction

Motor neuron diseases (MNDs) are a rather heterogeneous group of diseases, with either sporadic or genetic origin or both, all characterized by the progressive degeneration of motor neurons. All MNDs are primarily axonopathies of the motor neurons in which neuromuscular synapses are early targets of damage and death of motor neurons probably occurs following loss of the neuromuscular junctions [1]. MNDs may manifest as weakness, atrophy of muscles, difficulty in breathing, speaking, and swallowing, with symptoms and severity varying as a consequence of the different involvement of upper or lower motor neurons or both.
The most common and studied form in adults is Amyotrophic Lateral Sclerosis (ALS), followed by Progressive Bulbar Palsy (PBP), the rarer forms being Progressive Muscular Atrophy (PMA) and Primary Lateral Sclerosis (PLS). These conditions seem to form a continuum of diseases since only part of patients have a “pure” phenotype, while others with PBP or PLS eventually develop the widespread symptoms common to ALS [2]. In all these MNDs, onset of symptoms occurs mainly in people aged 40–70. Life expectancy is between 2 to about 5 years after onset in ALS and 6 months to 3 years in PBP, while pure PLS patients may have a normal or near-to-normal life duration. MNDs also include Spinal and Bulbar Muscular Atrophy (SBMA), in which age of onset and severity of manifestations vary from adolescence to old age, but longevity is usually not compromised. Infantile MNDs include Spinal Muscular Atrophy (SMA) with an infantile or juvenile onset and Lethal Congenital Contracture Syndrome (LCCS), causing prenatal death and thus being the most severe form of motor neuron disease.

3. Multifactoriality of MNDs: The Role of Altered Gene Expression

4. Altered Gene Expression, Mitochondrial Damage, and Oxidative Stress in MNDs: Which Are the Links?

5. A Unifying Mechanism for MNDs?

From what summarized above, it is tempting to speculate that indeed all MNDs are mainly forms of RNA dysmetabolisms.

Motor neurons seem to be exceedingly susceptible to defects in RNA transcription or processing; one appealing explanation is that they require that RNA is not only correctly transcribed and spliced, but also correctly transported along axons to neuromuscular junctions (NMJ).

While there is no clear demonstration of the presence of mRNAs at the NMJs yet, this process (at least in ALS) might resent from the known alterations in axonal transport that precedes onset of symptoms [102].

However, one form or the other of alteration of RNA expression may have different weight in different MNDs and, most importantly, RNA dysmetabolisms may be a primary event (for instance in SMA or in TDP43- and FUS/TLS-linked ALS) or dysregulation of components of the genetic machinery (the HATs/HDACs system, transcription factors, the splicing complex) may be secondary to oxidative stress or energy failure.

In turn, which step is the primary site of damage may dictate the severity of disease (age of onset, progression), and which cell type beside motor neurons is primarily affected may dictate the form of MND.

This field surely deserves further investigation aimed to the individuation of novel therapeutic approaches for MNDs.


http://www.hindawi.com/journals/ijcb/2012/908724/

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Olly
Posted: Thursday, June 21, 2012 9:48:01 AM

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Sodium phenylbutyrate prolongs survival and regulates expression of anti-apoptotic genes in transgenic amyotrophic lateral sclerosis mice.



Abstract

Multiple molecular defects trigger cell death in amyotrophic lateral sclerosis (ALS). Among these, altered transcriptional activity may perturb many cellular functions, leading to a cascade of secondary pathological effects. We showed that pharmacological treatment, using the histone deacetylase inhibitor sodium phenylbutyrate, significantly extended survival and improved both the clinical and neuropathological phenotypes in G93A transgenic ALS mice.

Phenylbutyrate administration ameliorated histone hypoacetylation observed in G93A mice and induced expression of nuclear factor-kappaB (NF-kappaB) p50, the phosphorylated inhibitory subunit of NF-kappaB (pIkappaB) and beta cell lymphoma 2 (bcl-2), but reduced cytochrome c and caspase expression.

Curcumin, an NF-kappaB inhibitor, and mutation of the NF-kappaB responsive element in the bcl-2 promoter, blocked butyrate-induced bcl-2 promoter activity.

We provide evidence that the pharmacological induction of NF-kappaB-dependent transcription and bcl-2 gene expression is neuroprotective in ALS mice by inhibiting programmed cell death.

Phenylbutyrate acts to phosphorylate IkappaB, translocating NF-kappaB p50 to the nucleus, or to directly acetylate NF-kappaB p50.

NF-kappaB p50 transactivates bcl-2 gene expression. Up-regulated bcl-2 blocks cytochrome c release and subsequent caspase activation, slowing motor neuron death. These transcriptional and post-translational pathways ultimately promote motor neuron survival and ameliorate disease progression in ALS mice. Phenylbutyrate may therefore provide a novel therapeutic approach for the treatment of patients with ALS.


PMID: 15934930 [PubMed - indexed for MEDLINE]

As a down stream factor:
Looking at the above is down regulation of bcl-2 and the cytochrome c release and subsequent caspase activation, common to most types of ALS in causing programmed motor neuron death or is there other pathways involved in different ALS subsets?


http://www.ncbi.nlm.nih.gov/pubmed/15934930

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That is the land of lost content,I see it shining plain,
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Olly
Posted: Friday, June 22, 2012 10:36:21 AM

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Table 1: Genes involved in MNDs.
________________________________________
Gene Protein MND Main known function
________________________________________
SOD1 Cu, Zn superoxide dismutase ALS1 Antioxidant enzyme
ALS2 Alsin ALS2 guanine nucleotide exchange factor for GTPases
SETX Senataxin ALS4 DNA/RNA metabolism and repair
SPG11 Spataxin ALS5 Neuron differentiation and axonal transport
FUS/TLS Fused in sarcoma ALS6 RNA binding protein
VAPB VAMP-associated protein B ALS8 Trafficking between the endoplasmic reticulum and Golgi apparatus
TDP-43 TAR-DNA-binding protein-43 ALS9 DNA- and RNA-binding protein
ANG Angiogenin ALS10 Angiogenesis in response to hypoxia; possibly RNA metabolism
FIG4 PI(3,5)P(2)5-phosphatase ALS11 Metabolism of phosphatidyl inositol bisphosphate and vesicle dynamic
OPTN Optineurin ALS12 Vesicular trafficking
nAChR Neuronal nicotinic acetylcholine receptor ALS Glutamatergic pathway
CHMP2B Charged multivesicular protein 2B ALS Chromatin-modifying protein/charged multivesicular body protein family
VCP Valosin-containing protein ALS Membrane trafficking, organelle biogenesis, maturation of ubiquitin-containing autophagosomes
DAO D-aminoacid oxidase ALS Oxidative deamination of D-aminoacid
UBQLN2 Ubiquilin2 ALS Ubiquitin-proteasome response
Sig-1R Sigma-1 receptor ALS ER chaperone, modulates calcium signaling through the IP3 receptor
C9ORF72 Unknown ALS Unknown
AR Androgen receptor SBMA Androgen receptor
SMN Survival Motor Neuron SMA RNA processing
GLE1 Nucleoporin GLE1 LCCS1 Export of mRNAs containing poly(A)
________________________________________

I have posted the above to help with identifying any commonality in the genetic processes.

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Olly
Posted: Friday, June 22, 2012 11:04:26 AM

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Listing of some known commonalities in MNDs:

Both sporadic and familial forms share a prominent role for mitochondrial damage and resulting oxidative stress.

Oxidative stress and mitochondrial dysfunction are obviously connected into a vicious cycle in which excess in ROS production may influence the functionality of the organelles, that in turn would produce excess ROS,


At the cellular level:

protein misfolding and aggregation

mitochondrial damage and energy deficit
Note that it has been reported that mitochondrial damage itself is a cause of modification in the abundance of selected gene splicing variants and that defective RNA metabolism seems to play a role also in SOD1-linked ALS and to descend directly from

mitochondrial stress

excitotoxicity,

calcium mishandling

altered control of gene expression
Note: Previous studies in ALS have revealed that most of the deregulated genes are involved in defense responses, cytoskeletal dynamics, protein degradation system, and mitochondrial dysfunction in neurons, while the insulin-like growth factor-1 receptor and the RNA-binding protein ROD1 are the most down regulated genes in glia

The pattern is altered also in muscle, in which many of deregulated genes are the same found in surgically denervated muscles, while others appear to be ALS-specific and include proteins clearly involved in the redox response (e.g., metallothionein-2 and thioredoxin-1).

Most of these proteins altered in ALS models, are involved in energy metabolism, cell stress response, protein degradation, and cytoskeleton stability.

As in other neurodegenerative conditions, alterations of transcription in MNDs may follow altered epigenetic control due to an unbalance between histone acetyl transferases (HATs) and histone deacetylases (HDACs, including sirtuins, SIRTs) activities


Into the heart, an air that kills, from yon far country blows.
What are those blue remembered hills, what sphires what farms are those.
That is the land of lost content,I see it shining plain,
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Olly
Posted: Friday, June 22, 2012 11:36:51 AM

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Heterogeneity in motoneuron disease

Recently, mutations in several genes have been
identified as primary causes for the degeneration of
motoneurons and their axons.

Strikingly, mutations in the same genes were associated with clinically different
motoneuron syndromes.

The identity of these genes also shed light on the mechanisms of motoneuron
degeneration and revealed that overlapping motoneuron phenotypes might be caused by heterogeneous molecular mechanisms.


Overall, these findings have challenged the diagnostic classification system set by
clinical judgement and triggered the notion of heterogeneity in motoneuron disease.


It will now be especially relevant to identify the mechanisms and principles that
motoneuron diseases have in common, as this will allow us to identify the most relevant therapeutic targets.

On the other hand, heterogeneity in motoneuron disease also implies that finding a monotherapy cure for motoneuron disease will be challenging and that pre-clinical
testing of therapeutic targets should not be limited to a single animal model
.

http://www.biochem.uthscsa.edu/~lafer/2008/Lambrechts-CarmelietNeurodegen07.pdf

Into the heart, an air that kills, from yon far country blows.
What are those blue remembered hills, what sphires what farms are those.
That is the land of lost content,I see it shining plain,
The happy highways where I went and cannot come again
Olly
Posted: Friday, June 22, 2012 12:19:30 PM

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Mitochondrial toxicity

By generating ATP and regulating cytoplasmic calcium, mitochondria are obviously of vital importance for neurons.

Mitochondria abundantly accumulate in synaptic terminals, which is thought to reflect the higher energy and calcium buffering needs of these tip structures.

Because mitochondria have a half-life of one month in the brain, an
intense axonal transport secures the supply of nuclearencoded proteins to the synapse for their assembly, whereas retrograde transport of mitochondria might serve
to replenish some of these building blocks.

It is thus not surprising, that mitochondrial damage or disruption of
their transport have been implicated in a wide variety of neurological diseases.

Mutations in genes encoded by the mitochondrial genome result in major neurological syndromes, but rarely in MNDs.

By contrast, mutations in nuclear-encoded mitochondrial genes cause MNDs.

Mice lacking paraplegin (SPG7), a gene mutated in HSP, develop a distal axonopathy
characterized by accumulation of mitochondria in axon tips [22].

Paraplegin proteolytically processes the mitochondrial ribosomal protein L32 (MRPL32), a subunit of mitochondrial ribosomes implicated in ribosome assembly [23].

Dominant-negative mutations in another nucleusencoded mitochondrial gene, mitofusin-2 (MFN2), cause various forms of CMT [13].

MFN2, which is located in the outer mitochondrial membrane, regulates fusion-and-fission
events that allow mitochondria to change their size and morphology according to the physiological state of the cell [13].

In neurons expressing disease-mutated forms of MFN2, the transport of mitochondria in axons was impaired.

This defect was not attributable to diminished ATP levels or oxidative respiration, thereby suggesting that abnormal mitochondrial trafficking itself caused axon
degeneration [24].


Damage to mitochondria also triggers motoneuron death in SOD1 mice.
Indeed, mitochondria become vacuolated early in the disease course [25].

Mutant SOD1 has also been found in the mitochondrial intermembrane space
and matrix, where it can interfere with proper mitochondrial functioning by: affecting the translocation machinery; generating toxic free radicals; forming aggregates, which
promote outer membrane vacuolization; and decreasing respiratory activity [25].

Also, mutant SOD1 can aggregate with the anti-apoptotic mitochondrial protein, B-cell lymphoma protein-2 (BCL-2) [26].

Together, all these effects might result in abnormal mitochondrial energy metabolism
and the release of apoptotic factors.

Intriguingly, both in rodent models and patient samples, mutant SOD1 is present in fractions enriched for mitochondria from affected, but not from unaffected tissues. Moreover, wildtype
SOD1 appears to be largely excluded from these mitochondrial preparations. Where exactly this dual selectivity comes from, remains however an unresolved key question.

http://www.biochem.uthscsa.edu/~lafer/2008/Lambrechts-CarmelietNeurodegen07.pdf

Into the heart, an air that kills, from yon far country blows.
What are those blue remembered hills, what sphires what farms are those.
That is the land of lost content,I see it shining plain,
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Olly
Posted: Friday, June 22, 2012 12:49:49 PM

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Returning to the down regulation of bcl-2 and the cytochrome c release and subsequent caspase activation, common to most types of ALS in causing programmed motor neuron death this may be relevant.


Caspase-3 Cleaves and Inactivates the Glutamate Transporter EAAT2

Abstract

EAAT2 is a high affinity, Na+-dependent glutamate transporter with predominant astroglial localization. It accounts for the clearance of the bulk of glutamate released at central nervous system synapses and therefore has a crucial role in shaping glutamatergic neurotransmission and limiting excitotoxicity.

Caspase-3 activation and impairment in expression and activity of EAAT2 are two distinct molecular mechanisms occurring in human amyotrophic lateral sclerosis (ALS) and in the transgenic rodent model of the disease.

Excitotoxicity caused by down-regulation of EAAT2 is thought to be a contributing factor to motor neuron death in ALS.

In this study, we report the novel evidence that caspase-3 cleaves EAAT2 at a unique site located in the cytosolic C-terminal domain of the transporter, a finding that links excitotoxicity and activation of caspase-3 as converging mechanisms in the pathogenesis of ALS.

Caspase-3 cleavage of EAAT2 leads to a drastic and selective inhibition of this transporter.

Heterologous expression of mutant SOD1 proteins linked to the familial form of ALS leads to inhibition of EAAT2 through a mechanism that largely involves activation of caspase-3 and cleavage of the transporter.

In addition, we found evidence in spinal cord homogenates of mutant SOD1 ALS mice of a truncated form of EAAT2, likely deriving from caspase-3-mediated proteolytic cleavage, which appeared concurrently to the loss of EAAT2 immunoreactivity and to increased expression of activated caspase-3.

Taken together, our findings suggest that caspase-3 cleavage of EAAT2 is one mechanism responsible for the impairment of glutamate uptake in mutant SOD1-linked ALS.

www.jbc.org/content/281/20/14076.full

Into the heart, an air that kills, from yon far country blows.
What are those blue remembered hills, what sphires what farms are those.
That is the land of lost content,I see it shining plain,
The happy highways where I went and cannot come again
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