Lipopolysaccharide-Induced Mitochondrial DNA Depletion

[Waverunner]

Very interesting since KDM tests and finds LPS in most of his patients.

http://www.ncbi.nlm.nih.gov/pubmed/21767162

Antioxid Redox Signal. 2011 Jul 18. [Epub ahead of print]

Lipopolysaccharide-Induced Mitochondrial DNA Depletion.
Choumar A, Tarhuni A, Lettron P, Reyl-Desmars F, Dauhoo N, Damasse J,
Vadrot N, Nahon P, Moreau R, Pessayre D, Mansouri A. INSERM, U773,
Centre de Recherche Biomdicale Bichat Beaujon CRB3 , Paris, France .

Abstract Hepatic energy depletion has been described in severe sepsis,
and lipopolysaccharide (LPS) has been shown to cause mitochondrial DNA
(mtDNA) damage. To clarify the mechanisms of LPS-induced mtDNA damage
and mitochondrial alterations, we treated wild-type (WT) or transgenic
manganese superoxide dismutase-overerexpressing (MnSOD(+++)) mice with
a single dose of LPS (5?mg/kg). In WT mice, LPS increased
mitochondrial reactive oxygen species formation, hepatic inducible
nitric oxide synthase (NOS) mRNA and protein, tumor necrosis factor-
alpha, interleukin-1 beta, and high-mobility group protein B1
concentrations. Six to 48?h after LPS administration (5?mg/kg), liver
mtDNA levels, respiratory complex I activity, and adenosine
triphosphate (ATP) contents were decreased. In addition, LPS increased
interferon-? concentration and decreased mitochondrial transcription
factor A (Tfam) mRNA, Tfam protein, and mtDNA-encoded mRNAs.
Morphological studies showed mild hepatic inflammation. The LPS (5?mg/
kg)-induced mtDNA depletion, complex I inactivation, ATP depletion,
and alanine aminotransferase increase were prevented in MnSOD(+++)
mice or in WT mice cotreated with 1400W (a NOS inhibitor), (2-(2,2,6,6-
tetramethylpiperidin-1-oxyl-4-ylamino)-2-oxoethyl)triphenylphosphonium
chloride, monohydrate (a superoxide scavenger) or uric acid (a
peroxynitrite scavenger). The MnSOD overexpression delayed death in
mice challenged by a higher, lethal dose of LPS (25?mg/kg). In
conclusion, LPS administration damages mtDNA and alters mitochondrial
function. The protective effects of MnSOD, NOS inhibitors, and
superoxide or peroxynitrite scavengers point out a role of the
superoxide anion reacting with NO to form mtDNA- and protein-damaging
peroxynitrite. In addition to the acute damage caused by reactive
species, decreased levels of mitochondrial transcripts contribute to
mitochondrial dysfunction.
PMID:21767162

 

[alex3619]

Hi, KDM does not just find LPS in patients, he finds it proportional to severity of symptoms. Also, this is potentially the driving force behind the NO/ONOO issues and much of our oxidative stress. I wonder if it can also explain our mitochondrial dysfunction - and why it differs a little patient by patient, it may depend on which mitochondrial genes are damaged. Bye, Alex

 

[Waverunner]

Hi, KDM does not just find LPS in patients, he finds it proportional to severity of symptoms. Also, this is potentially the driving force behind the NO/ONOO issues and much of our oxidative stress. I wonder if it can also explain our mitochondrial dysfunction - and why it differs a little patient by patient, it may depend on which mitochondrial genes are damaged. Bye, Alex

Yes, the genes probably have an influence on how much our mitochondria get compromised. The most important question however seems to be what causes the LPS in our blood. Bacteria probably but why do we have these bacteria and why do they or a our immune system malfunction? Antibiotics are no solution, they can make things a lot better but the bacteria return sooner or later so what makes them grow so good?

 

[alex3619]

Hi waverunner, everyone has masses of LPS in the gut. The gut is supposed to detox it, and the liver picks up the rest. Therefore both the gut and liver detox have failed in us. So the question is, why have they failed? I don't know much/enough about specific LPS detox, but it would not surprise me if it is glutathione dependent. This could be feeback loops (eg methylation), cytokines or a virus infection as I see it, although other specific toxins may play a role. Bye, Alex

 

[Waverunner]

Hi waverunner, everyone has masses of LPS in the gut. The gut is supposed to detox it, and the liver picks up the rest. Therefore both the gut and liver detox have failed in us. So the question is, why have the failed? I don't know much/enough about specific LPS detox, but it would not surprise me if it is glutathione dependent. This could be feeback loops (eg methylation), cytokines or a virus infection as I see it, although other specific toxins may play a role. Bye, Alex

Yes, one can probably break down the cause to a virus, methylation or a genetic defect. It's still sad however, that we have no answer also we probably could have, if more scientists would look into this field and if it would receive more attention.

 

[fla]

Can mitochondrial DNA depletion be reversed? Really hope so otherwise how can we find a cure for M.E.?

 

[rwac]

The answer is yes.

http://www.ncbi.nlm.nih.gov/pubmed/14640394

Uridine abrogates mitochondrial toxicity related to nucleoside analogue reverse transcriptase inhibitors in HepG2 cells.

OBJECTIVE:
To assess in vitro if uridine may be suitable to prevent or treat mitochondrial toxicity related to nucleoside analogue reverse transcriptase inhibitors (NRTIs).

METHODS:
Human HepG2-hepatocytes were exposed to NRTIs with or without uridine for 25 days. Cell growth, lactate production, intracellular lipids, mitochondrial DNA (mtDNA) and the ratio between the respiratory chain components COX II (mtDNA-encoded) and COX IV (nuclear-encoded) were measured.

RESULTS:
HepG2 cells exposed to zalcitabine (177 nM) without uridine developed a severe depletion of mtDNA (to 8% of wild-type mtDNA levels), resulting in a decline of cell proliferation and COX II levels, with increased lactate and lipid accumulation. Uridine fully abrogated the adverse effects of zalcitabine on hepatocyte proliferation and normalized lactate synthesis, intracellular lipids and COX II levels by adjusting mtDNA levels to about 65% of NRTI-unexposed control cells. This effect was dose-dependent, with a maximum at 200 microM of uridine. Uridine also rapidly and fully restored cell function when added to cells with established mitochondrial dysfunction (zalcitabine for 15 days) despite continued zalcitabine exposure. Uridine also normalized cell proliferation in HepG2 cells exposed to 36 microM of stavudine and protected HepG2-cells exposed to 7 microM of zidovudine + 8 microM of lamivudine (pyrimidine analogues), but failed to improve cell function or mtDNA in cells exposed to 11.8 or 118 microM of didanosine (a purine analogue).

CONCLUSIONS:
The pyrimidine precursor uridine may attenuate the mitochondrial toxicity of antiretroviral pyrimidine NRTIs in vitro, and its supplementation may represent a promising strategy in the prevention or treatment of mitochondrial toxicities in HIV-infected patients.

Uridine in the prevention and treatment of NRTI-related mitochondrial toxicity.
Walker UA, Venhoff N.

Abstract
Long-term side effects of antiretroviral therapy are attributed to the mitochondrial (mt) toxicity of nucleoside analogue reverse transcriptase inhibitors (NRTIs) and their ability to deplete mtDNA. Studies in hepatocytes suggest that uridine is able to prevent and treat mtDNA depletion by pyrimidine NRTls [zalcitabine (ddC) and stavudine (d4T)] and to fully abrogate hepatocyte death, elevated lactate production and intracellular steatosis. Uridine was also found to improve the liver and haematopoietic toxicities of zidovudine (AZT), which are unrelated to mtDNA depletion, and to prevent neuronal cell death induced by ddC. Most recently, uridine was found to prevent the onset of a lipoatrophic phenotype (reduced intracellular lipids, increased apoptosis, mtDNA depletion and mt depolarization) in adipocytes incubated long-term with d4T and AZT. Various steps of mt nucleoside utilization may be involved in the protective effect, but competition of uridine metabolites with NRTIs at polymerase y or other enzymes is a plausible explanation. Pharmacokinetic studies suggest that uridine serum levels can be safely increased in humans to achieve concentrations which are protective in vitro (50-200 microM). Uridine was not found to interfere with the antiretroviral activity of NRTIs. Mitocnol, a sugar cane extract which effectively increases uridine in human serum, was beneficial in individual HIV patients with mt toxicity and is now being tested in placebo-controlled randomized trials. Until these data become available, the risk-benefit calculation of using uridine should be individualized. The current safety data justify the closely monitored use of uridine in individuals who suffer from mt toxicity but who cannot be switched to less toxic NRTIs.

 

[rwac]

And the best source of uridine is probably from an RNA supplement.

 

[Marco]

There is a French immunologist called Gregoire Cozon based in Lyon who reports detecting antigens to Candida albicans or Staphylococcus aureus in 50 to 60% of patients and hypothesises that we develop a abnormal immune reaction to these common microbes following vaccination or viral infection.

Unfortunately I can't find the source document but if you Google it is described on a French blog that can be google translated.

http://sfc-fibro.over-blog.com/pages/III_LES_EXAMENS_DE_SECONDES_INTENTIONS-1464549.html#

Staphylococcus aureus has also been shown to disrupt the mitochondria.

Guptal et al use LPS to induce fatigue in mice as an animal model of ME/CFS and have shown that various polyphenols (e.g. curcumin) reduce fatigue; and attenuate oxidative stress and TNF alpha :

http://www.ncbi.nlm.nih.gov/pubmed/19159825

Getting back to Staphylococcus aureus, SA sepsis causes oxidative stress and mitochondrial damage but this is reversed by mitochondrial biogenesis activated partly by coactivator peroxisome proliferator-activated receptor ? coactivator-1? (a PPAR) :

http://respiratory.publishingtechnology.com/content/article/10.1164/rccm.200701-161OC

PPAR activity can be stimulated by a range of compounds e.g. Quercetin :

http://www.ncbi.nlm.nih.gov/pubmed/19211721

So perhaps there may be an opportunity to repair acquired mito dysfunction?

 

[alex3619]

Can mitochondrial DNA depletion be reversed? Really hope so otherwise how can we find a cure for M.E.?

Hi fla, I have a different answer to that given here, a better solution. I don't think it will be possible to reverse the DNA damage as suggested, not in our lifetimes. The solution is simpler than that. If all our mitochondria were damaged, we would be dead. We have many many healthy mitochondria still. What we need is a way to force those mitochondria to multiply, With more healthy mitochondria, the damaged mitochondrial DNA wont mean as much. DHEA is one substance that does this, but I am sure there are others. So the situation is not irreversible, nor as tricky as trying to repair damaged DNA.

Bye
Alex

 

[rwac]

Yes, PQQ should help with generating new mitochondria. Helps me quite a bit.

 

[mellster]

And the best source of uridine is probably from an RNA supplement.

Hey rwac, something like brewers yeast for example? cheers

 

[rwac]

Hey rwac, something like brewers yeast for example? cheers

Absolutely, brewer's yeast would work. Unfortunately it triggers asthma attacks for me, so I switched to the lef RNA supplement.

 

[Waverunner]

I wanna throw in a comment: http://www.longecity.org/forum/topic/9534-anyone-have-experiences-with-rna-supplements/

"I seriously doubt ingesting RNA would lead to the protein it initially coded for being expressed in any tissues. My reasoning is that RNA is generally speaking fairly unstable. If you wanted to extract RNA from cells in the lab (for use in a micro-array analysis for example) it is done on wet ice and with a cocktail of inhibitors. Even without RNase's it can break down. DNA is more stable because it has one less hydroxy group on the (deoxy)ribose sugar."

"Be careful when using RNA/DNA supplements. The breakdown of these product causes excessive Uric acid which causes gout. I took the LEF RNA capsules for a year and my Uric acid level went above 7 or 8 causing severe joint pain. Stopped taking the RNA and after 12 months I was back to normal. I don't normally test for Uric Acid on my blood test, it was only after my joints became a pain that I found the culprit. "

 

[alex3619]

"Be careful when using RNA/DNA supplements. The breakdown of these product causes excessive Uric acid which causes gout. I took the LEF RNA capsules for a year and my Uric acid level went above 7 or 8 causing severe joint pain. Stopped taking the RNA and after 12 months I was back to normal. I don't normally test for Uric Acid on my blood test, it was only after my joints became a pain that I found the culprit. "

This comment is technically correct, but not really so important if you have ME. The purine breakdown product is indeed uric acid, but its a highly protective antioxidant. For us this would be a good thing unless, as in gout, the acid crystallizes. Bye, Alex

 

[Waverunner]

K, thanks, Alex.

 

[Marco]

Uridine is contained in Beer!

Add some dark chocolate and we could be in business :

EPICATECHIN ENHANCES FATIGUE RESISTANCE AND OXIDATIVE CAPACITY IN MOUSE MUSCLE

http://jp.physoc.org/content/early/2011/07/24/jphysiol.2011.209924.abstract

 

[aprilk1869]

I just came across this, you can download the full pdf here: http://www.spandidos-publications.c...ype=article&article_id=ijmm_22_6_731&item=PDF

Essential nutrients suppress inflammation by modulating key inflammatory gene expression

We investigated the effects of a nutrient mixture (NM) consisting of ascorbic acid, quercetin, naringenin, hesperetin, tea catechins, lysine, proline, arginine and N-acetylcysteine on experimental in vivo and in vitro inflammation triggered by bacterial lipopolysaccharide (LPS). BALB/c mice (n=36) were administered NM (200 mg/kg BW) or ibuprofen (20 mg/kg BW) for two weeks. Blood plasma, collected three hours after a single intraperitoneal injection with LPS (1 mg/kg BW), was analyzed with 14 cytokine microarray. LPS inflammatory effects were analyzed in human U937 macrophages by cytokine release, cyclooxygenase (COX) enzymatic activity, COX protein expression (Western blot analysis), specific mRNA levels (RT-PCR), and nuclear factor ?? (NF??) activation (phosphorylated p65 immunoassay). Nutrient supplementation in mice altered the LPS-induced cytokine response in a manner similar to ibuprofen (r=0.4157, p=0.139). Cytokine response to LPS in cultured macrophages was similar to the in vivo study (r=0.718, p=0.023). NM inhibited COX-2 enzymatic activity, and COX-2 and pro-inflammatory cytokine protein expression levels were downregulated by NM at the transcription level complementing a blockade in NF?? activation. NM demonstrated strong beneficial effects on the experimental inflammation by targeting multiple responsible mechanisms in the complex process involved in the inflammatory reaction to pathogens.

http://www.spandidos-publications.com/ijmm/22/6/731

 

[alex3619]

Dr Andriya Martinovic

Hi aprilk1869, the nutrient mixture abstract is interesting. Something similar was being used in the early to mid 90s to treat CFS but the researcher (my doctor at the time) could not get funding and had trouble publishing papers. In the end he was forced to stop treating his patients due to this being experimental medicine and not common practice. His success rate was ignored.

Just to be clear, his treatment was similar but also different in that he used titration of key substrates to block COX2 hyperutilization.

Bye
Alex

 

[Marco]

A little initial digression with this anecdote if I may.

When we first moved to France, one of the first things we noticed was that supermarkets were not the same 'one stop shops' that they are in the UK. For example, you can't buy cigarettes - you have to go to a tobacconist and you can't buy any health products over and above simple vitamin supplements or natural remedies. Any painkillers/cold flu remedies etc require a trip to the pharmacy. While this can be a little bit of a pain, I suppose it does help ensure that local town centres retain some commerce.

Anyway, the first time one of came down with a seasonal cold or bug we went to the pharmacy and they gave us a product called Mucomyst - for drying up mucus. We have found this so useful we call it 'magic potion'. It seems to work on most things whether a cold, flu, bug or just generally feeling a little off colour (my wife doesn't have ME by the way).

For me it has additional benefits. I'm very intolerant of high temperature and heat can frequently cause PEM. A sachet of Mucomyst can prevent this or get me back on my feet. I've also found that a sachet taken in advance can prevent PEM if I have any prolonged physical chores to do.

Its only lately that I thought of trying to find out what was in our 'magic potions'. Its acetylcysteine.

Apparently acetylcysteine is both a powerful antioxidant plus a glutathione precursor (for this reason its used as a treatment for paracetamol overdose).

Maes also reports success in using acetylcysteine plus glutamine and zinc to treat ME/CFS leaky gut :

http://www.ncbi.nlm.nih.gov/pubmed/19112401


All anecdotal of course (my bits) but it works for me and its cheap as chips.