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Study: Mitomycin C & pulsed ceftriaxone kill lyme persister cells

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Posted 10/21/2017 4:03 AM (GMT 0)
http://aac.asm.org/content/59/8/4616.full

Somebody said...


Borrelia burgdorferi, the Causative Agent of Lyme Disease, Forms Drug-Tolerant Persister Cells

Bijaya Sharmaa, Autumn V. Browna, Nicole E. Matlucka, Linden T. Hub and Kim Lewisa

ABSTRACT

Borrelia burgdorferi is the causative agent of Lyme disease, which affects an estimated 300,000 people annually in the United States. When treated early, the disease usually resolves, but when left untreated, it can result in symptoms such as arthritis and encephalopathy. Treatment of the late-stage disease may require multiple courses of antibiotic therapy. Given that antibiotic resistance has not been observed for B. burgdorferi, the reason for the recalcitrance of late-stage disease to antibiotics is unclear. In other chronic infections, the presence of drug-tolerant persisters has been linked to recalcitrance of the disease. In this study, we examined the ability of B. burgdorferi to form persisters. Killing growing cultures of B. burgdorferi with antibiotics used to treat the disease was distinctly biphasic, with a small subpopulation of surviving cells.

Upon regrowth, these cells formed a new subpopulation of antibiotic-tolerant cells, indicating that these are persisters rather than resistant mutants.

The level of persisters increased sharply as the culture transitioned from the exponential to stationary phase. Combinations of antibiotics did not improve killing.Daptomycin, a membrane-active bactericidal antibiotic, killed stationary-phase cells but not persisters.

Mitomycin C, an anticancer agent that forms adducts with DNA, killed persisters and eradicated growing and stationary cultures of B. burgdorferi.

Finally, we examined the ability of pulse dosing an antibiotic to eliminate persisters. After addition of ceftriaxone, the antibiotic was washed away, surviving persisters were allowed to resuscitate, and the antibiotic was added again.

Four pulse doses of ceftriaxone killed persisters, eradicating all live bacteria in the culture.

Posted 10/21/2017 4:53 AM (GMT 0)
https://www.ncbi.nlm.nih.gov/pubmed/25806811

Somebody said...


Drug combinations against Borrelia burgdorferi persisters in vitro: eradication achieved by using daptomycin, cefoperazone and doxycycline.

Abstract

Although most Lyme disease patients can be cured with antibiotics doxycycline or amoxicillin using 2-4 week treatment durations, some patients suffer from persistent arthritis or post-treatment Lyme disease syndrome. Why these phenomena occur is unclear, but possibilities include host responses, antigenic debris, or B. burgdorferi organisms remaining despite antibiotic therapy. In vitro, B. burgdorferi developed increasing antibiotic tolerance as morphology changed from typical spirochetal form in log phase growth to variant round body and microcolony forms in stationary phase. B. burgdorferi appeared to have higher persister frequencies than E. coli as a control as measured by SYBR Green I/propidium iodide (PI) viability stain and microscope counting. To more effectively eradicate the different persister forms tolerant to doxycycline or amoxicillin, drug combinations were studied using previously identified drugs from an FDA-approved drug library with high activity against such persisters. Using a SYBR Green/PI viability assay, daptomycin-containing drug combinations were the most effective.

Of studied drugs, daptomycin was the common element in the most active regimens when combined with doxycycline plus either beta-lactams (cefoperazone or carbenicillin) or an energy inhibitor (clofazimine).

Daptomycin plus doxycycline and cefoperazone eradicated the most resistant microcolony form of B. burgdorferi persisters and did not yield viable spirochetes upon subculturing,

suggesting durable killing that was not achieved by any other two or three drug combinations. These findings may have implications for improved treatment of Lyme disease, if persistent organisms or detritus are responsible for symptoms that do not resolve with conventional therapy. Further studies are needed to validate whether such combination antimicrobial approaches are useful in animal models and human infection.

Posted 10/21/2017 5:06 AM (GMT 0)
http://news.northeastern.edu/2015/06/researchers-discovery-may-explain-difficulty-in-treating-lyme-disease/

Somebody said...
In addition to identifying the presence of these persister cells, Lewis’ team also presented two methods for wiping out the infection—both of which were successful in lab tests. One involved an anti-cancer agent called Mitomycin C, which completely eradicated all cultures of the bacterium in one fell swoop. However, Lewis stressed that, given Mitomycin C’s toxicity, it isn’t a recommended option for treating Lyme disease, though his team’s findings are useful to helping to better understand the disease.

The second approach, which Lewis noted is much more practical, involved pulse-dosing an antibiotic to eliminate persisters.

The researchers introduced the antibiotic a first time, which killed the growing cells but not the dormant persisters. But once the antibiotic washed away, the persisters woke up, and before they had time to restore their population the researchers hit them with the antibiotic again. Four rounds of antibiotic treatments completely eradicated the persisters in a test tube.

“This is the first time, we think, that pulse-dosing has been published as a method for eradicating the population of a pathogen with antibiotics that don’t kill dormant cells,” Lewis said.

“The trick to doing this is to allow the dormant cells to wake up.”

He added: “This gives you an idea that you could, in principle, establish a similar regiment for treating patients for this and other chronic diseases.”

Posted 10/21/2017 5:54 AM (GMT 0)
it is an old study, proven to be inaccurate by the zhang team. Zhang's team has a superior borrelia growth strategy, it lets them grow much longer than Lewis' team and thus u can see what happens longer term. He tried Mitomycin C and find it DOES NOT sterilize the bacteria

Eradication of Biofilm-Like Microcolony Structures of Borrelia burgdorferi by Daunomycin and Daptomycin but not Mitomycin C in Combination with Doxycycline and Cefuroxime.
/www.ncbi.nlm.nih.gov/pubmed/26903956

besides that, Mitomycin C is a very toxic anti cancer drug, and it does not cross the BBB,

"However, an intact blood-brain barrier in the brain and spinal cord prevented mitomycin C from producing adverse effects in the central nervous system (Schwartz and Philips, 1961; Lee et al., 2006b)"

...so it is NOT a good treatment candidate for lyme.
Posted 10/21/2017 6:16 AM (GMT 0)
So, what could be worse than long-term antibiotics?

Answer: Chemotherapy. (IMO)
Posted 10/21/2017 8:53 AM (GMT 0)
and i believe Zhangs team, in conjunction with harvard showed Oil of oregano was more effective than Daptomycin for persisters
Posted 10/21/2017 3:40 PM (GMT 0)

dbwilco said...
and i believe Zhangs team, in conjunction with harvard showed Oil of oregano was more effective than Daptomycin for persisters

which is somewhat irrelevant

you will never be able to reach MBC or even MIC in the human bloodstream with O of O. You would likely die first

Somebody said...
In microbiology, the minimum inhibitory concentration (MIC) is the lowest concentration of a chemical which prevents visible growth of a bacterium. This is in difference to the minimum bactericidal concentration (MBC) which is the concentration resulting in microbial death as defined by the inability to re-culture bacteria. The closer the MIC is to the MBC, the more bactericidal the compound.

IF he could prove you could reach MBC with O of O in the human blood stream, then great. otherwise its like saying "bleach kills borrelia in a petri dish betther than antibiotics!"

ok great, who cares? you cant drink bleach.

Post Edited (Lapis_29) : 10/21/2017 9:44:07 AM (GMT-6)

Posted 10/21/2017 3:43 PM (GMT 0)
liposomal oil of oregano is what many LLMds use from a compounding pharmacy
Posted 10/21/2017 3:45 PM (GMT 0)

dbwilco said...
liposomal oil of oregano is what many LLMds use from a compounding pharmacy

almost certainly will not work

I mean will it help a bit? sure probably, but you cant take enough to really make a dramatic difference in my opinion
Posted 10/21/2017 3:58 PM (GMT 0)
is your opinion based on using it?? im on it, and feel an effect on my neurolyme
Posted 10/21/2017 4:06 PM (GMT 0)

dbwilco said...
is your opinion based on using it?? im on it, and feel an effect on my neurolyme

then you should keep using it. I mean if it works, its awesome

I am just saying it likely will never be a stand alone treatment.
Posted 10/21/2017 4:08 PM (GMT 0)

Lapis_29 said...

you will never be able to reach MBC or even MIC in the human bloodstream with O of O. You would likely die first

Here we go again, Girlie will tell me again i need to check my tone sad

i am sorry to say Lapis_29, but even if you are a doctor, i tend to believe mr Zhang and not you (no hard feelings, it's simply because he happens to be a microbiologist specialized in persistent infections, like TB, so quite qualified, i might add). And Zhang says something else than you about oil of oregano/carvacrol:

"In the rat model, the calculated LD50 of carvacrol is 471.2 mg/kg (30). We noticed that the 0.05% of carvacrol used here, which is equivalent to 0.48 µg/ml or 3.2 µM and completely eradicated B. burgdorferi stationary phase cells in subculture (Figure 3), is lower than the peak plasma concentration (3.65 µg/ml) in the swine study (28). Clinically, an antimicrobial agent that penetrates the blood–brain barrier as well as the persistent Borrelia organisms do, would be an ideal candidate for further study (31). Oil of Oregano is a good candidate for this consideration given that its main active ingredient—carvacrol is a phenolic monoterpenoid that is fat-soluble and found in animal studies to have blood–brain barrier penetration (32). These findings favor the application of carvacrol in future treatment studies. Importantly, carvacrol seems to be more active than Dap, the most active persister drug against B. burgdorferi (14, 15)."

Anyway, please read the full study, then we can comment or tell people our educated guesses.
/www.frontiersin.org/articles/10.3389/fmed.2017.00169/full

If we don't read up things, then our guesses are just guesses, uneducated and not useful. We here try to find a way to get well together. For this, an important part is reading a ton of information, ideally peer reviewed articles. It is boring, time consuming, and in 99% cases completely useless because the studies that really matter are few and u need to search for them constantly ...

So please, let's stick to the facts... Right now it looks like carvacrol is a really good chemical to destroy borrelia everywhere in your body. The problem is oil of oregano is not the same thing as doxy, it is produced by a plant not a factory, so its potency can vary. It must be transformed into a real drug with controlled ingredients and potency, if we are to have a chance of it to cure anyone ....

Thanks and again ... apologies for my tone. I read A LOT of papers in the last 4 years, wasted a lot of time with this, so i might be too emotionally invested into this...
Posted 10/21/2017 4:13 PM (GMT 0)

mpost said...

Lapis_29 said...

you will never be able to reach MBC or even MIC in the human bloodstream with O of O. You would likely die first


"In the rat model, the calculated LD50 of carvacrol is 471.2 mg/kg (30). We noticed that the 0.05% of carvacrol used here, which is equivalent to 0.48 µg/ml or 3.2 µM and completely eradicated B. burgdorferi stationary phase cells in subculture (Figure 3),

so based on that you think you could reach MBC with carvacrol?
Posted 10/21/2017 4:50 PM (GMT 0)

Lapis_29 said...

so based on that you think you could reach MBC with carvacrol?

just take 40 mins and read the whole study, or at least the results and discussion sections. He goes in great detail in the Discussion section, about carvacrol, toxicity, etc...
Posted 10/21/2017 5:15 PM (GMT 0)
mpost -

You may have a "tone" problem ;) but sometimes I do too.

And it's not that you're wrong, necessarily. Or that the studies are wrong.

It's that current science - even today - understands a fraction of the complexity of Bb, its impact on immune function of a society that is experiencing broad interference with normal immunity for several other reasons, within a toxic environment.

There are just way too many variables in anyone treating Lyme for people ---even the best microbiologists and researchers--- to make "absolute" statements.

The key for each and every one of us is to figure out what works for us. Your research is great and is helping you form your own protocol and it's also great that you're sharing your research.

It's probably most helpful, however, to let the research speak for itself, offer your opinions about what it means for YOUR unique situation and share how you're utilizing the research to inform your decisions. And avail yourself for questions and engage in dialogue.

-p
Posted 10/21/2017 5:34 PM (GMT 0)
Pirouette,
i agree with everything you said.

however for me the fact that carvacrol is so powerful against borrelia , is an eye opener. i knew for some time, as many members of the forum here, that oil of oregano is good against lyme. this is not news. but so is doxy, mino, ceftin, ceftriaxone, amoxicilin, andrographis, etc etc etc. the list is immense.

however there are not so many ingredients that
a) cross the BBB
b) get absorbed orally
c) reach excess of MIC levels
d) completely kill borrelia with persisters, biofilms, reproductive form ...
e) do all the above without reaching toxic levels that kill the host - this has to be yet proven but it is a good guess

careful, carvacrol is toxic in large amounts. So as Pirouette said, let the researchers do their work. Test this on mice first. Cure a few mice from chronic lyme, then we can take it without ruining our liver/etc.

in the mean time, take your doxy smile

it is just good to feel progress is being made...
Posted 10/21/2017 5:56 PM (GMT 0)
my doctor is also on the forefront of new research (currently getting funding for animal testing right now along with another well known name in the lyme community), and has told me that oil of oregano has also been shown to be effective in the testtube against brucella (a tougher cousin of bart)....he said the white blood cells "eat" the liposomal form, so that he finds it may be effective against bart as well, in conjunction with the rest of his protocol...from personal experience it DEF crosses the BBB

mpost, thank you, i personally find you to be the most beneficial poster on the forum, on the cutting edge of new research
Posted 10/21/2017 6:23 PM (GMT 0)
Now I'm curious who your doc is dbwilco! Great info here. I took Gaia oil of oregano and feel nothing. I'm guessing the dose is not strong enough. Looking forward to more studies being released. And I agree, I find mpost's posts (lol) very helpful. I can't get enough of the medical research!
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