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Re: Info about Lyme round bodies (cysts)

Posted: Sat 31 Dec 2011 19:41
by Henry
Margarita: There is no evidence to support such statements made by ILADS physicians. These structures will disappear in time; no antibiotics --given in any amount-- are going to make that happen faster. More important, there is NO evidence that these structures have any clinical relevance.

Re: Info about Lyme round bodies (cysts)

Posted: Sat 31 Dec 2011 21:05
by X-member
Removed post!

Re: Info about Lyme round bodies (cysts)

Posted: Sun 1 Jan 2012 2:22
by ChuckG
The pdf has been on my hard drive since Sep 13, 2010. I am agnostic on the question of the cyst form.

http://www.ncbi.nlm.nih.gov/pubmed?term ... 2010658658

Microbiology. 2000 Jan;146 ( Pt 1):119-27.
Serum-starvation-induced changes in protein synthesis and morphology of Borrelia burgdorferi.
Alban PS, Johnson PW, Nelson DR.
Source
Department of Biochemistry, Microbiology, and Molecular Genetics, University of Rhode Island, Kingston 02881, USA.

Abstract
It has been demonstrated previously that motile Borrelia burgdorferi cells transform into non-motile cyst-forms when incubated for several weeks in BSKII (a complex medium) lacking rabbit serum. B. burgdorferi cells cannot synthesize fatty acids de novo and serum is thought to provide a source of fatty acids and lipids. When B. burgdorferi cells were serum-starved in defined RPMI medium, -90% of the cells formed spherical cysts within 48 h. Cyst formation was inhibited by tetracycline. Cyst opening and recovery of vegetative cells was rapidly induced by the addition of either BSKII or rabbit serum. The percentage of viable cells recovered from cysts ranged from 2.9% to 52-5%. Viability was inversely proportional to cyst age. Protein synthesis by B. burgdorferi during serum starvation was examined by labelling cells with Tran35S-Label and analysing the labelled proteins by two-dimensional gel electrophoresis and fluorography. The synthesis of over 20 proteins was induced during serum starvation. Western blots of proteins from vegetative cells and cysts probed with sera from either B. burgdorferi-infected humans or monkeys revealed that several cyst proteins were antigenic. These data suggest that cells of B. burgdorferi, although possessing a small genome and extremely limited biosynthetic capabilities, rapidly respond to conditions of serum starvation by inducing changes in protein synthesis and cell morphology. This study may help explain how cells of B. burgdorferi can survive periods of nutrient deprivation in different hosts and host tissues.

PMID: 10658658 [PubMed - indexed for MEDLINE] Free full text

"... Within 24 h, cells starved of serum were completely non-motile and 30–40 % had begun to encyst. After 48 h incubation in RPMI,, 90 % of serum-starved cells had formed cysts (Fig. 1)."

See Fig. 1C. Cyst is a single spirochete coiled up to ≈ 2µm.

Re: Info about Lyme round bodies (cysts)

Posted: Sun 1 Jan 2012 2:36
by Camp Other
Carina:

Can you cite any references which state that these forms (cell-wall or cell-membrane deficient variants, L forms, sphaeroplasts, and/or protoplasts - Henry is correct; "cyst" is not the correct scientific term for this form of Bb) last for a particular period of time in vivo? In animal or human models? And is there evidence that they are somehow more resistant to antibiotics in vivo than spirochetal forms are?

I know there is much discussion in various Lyme disease support groups of how Borrelia burgdorferi can survive for long times in this sort of round form when exposed to adverse conditions - but I agree with Henry that I don't think they are as relevant as others seem to think they are - in part because their composition is essentially the same as the spirochete's.

As an aside, If you want to look at two things which may be of greater interest in solving the persisting symptoms puzzle, I suggest looking into the below:

1) Whether or not Borrelia can go into a semi-dormant or dormant state after antibiotic exposure. The shape or form really isn't important. (see mRNA)

2) How the immune system responds to infection in lymph nodes and other parts of the host - and whether or not this same event as captured in Barthold's studies on mice occurs in humans.

Beyond these two items, more new research is needed to pinpoint the cause of persisting symptoms in patients after they have already had antibiotic treatment.

The more research I read, the more I'm hypothesizing that genetics plus immune system dysregulation play a role in patients' persisting symptoms.

Having a particular strain or strains of Borrelia to begin with could influence choice of antibiotic treatment if known - there is evidence that test result presentation is related to strain (see: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2772371/) and also, there are European studies on in vitro antibiotic choice which have pointed out that certain strains respond to different antibiotics differently.

This may affect patients in that tests may not reflect reality and/or some patients may not be getting the most effective antibiotic treatment for the strain with which they are infected - but this is speculation on my part. I think that strain is only a small part of the story here.

Re: Info about Lyme round bodies (cysts)

Posted: Sun 1 Jan 2012 2:42
by X-member
Hi CO!

The only thing I did (in another topic) was to call a "round body" in a picture taken on my blood outside the body for a cyst! And this Henry didn't like!

The rest of the discussion is actually not mine! :D

And the only thing I wanted from Henry, was the right term for the "round body".

Re: Info about Lyme round bodies (cysts)

Posted: Sun 1 Jan 2012 2:51
by X-member
CO you wrote:
I think that strain is only a small part of the story here.
I agree!

Re: Info about Lyme round bodies (cysts)

Posted: Sun 1 Jan 2012 2:58
by Camp Other
ChuckG:

Nice citation, to point out Alban PS, Johnson PW, and Nelson DR.'s use of the word, "cyst". I think they are using the incorrect word too, though.

When I read something like your cited passages, I call my attention to the items I bold below and ask questions:
"Cyst formation was inhibited by tetracycline. Cyst opening and recovery of vegetative cells was rapidly induced by the addition of either BSKII or rabbit serum. The percentage of viable cells recovered from cysts ranged from 2.9% to 52-5%. Viability was inversely proportional to cyst age. Protein synthesis by B. burgdorferi during serum starvation was examined by labelling cells with Tran35S-Label and analysing the labelled proteins by two-dimensional gel electrophoresis and fluorography. The synthesis of over 20 proteins was induced during serum starvation. Western blots of proteins from vegetative cells and cysts probed with sera from either B. burgdorferi-infected humans or monkeys revealed that several cyst proteins were antigenic."
If "cyst" formation is inhibited by tetracycline, would long term antibiotic use using tetracycline (possibly doxycycline) prevent "cysts" from forming at all? If the "cyst" form is so important in establishing chronic infection as has been speculated by some, then why not just give people the drugs which inhibit their formation?

"Viability was inversely proportional to 'cyst' age": This means the longer a "cyst" is around, the less viable it is. As in, it's not likely to revert and survive as a spirochete. Does this imply that if adverse conditions occur inside the host that "cysts" - if they do occur - will deteriorate over time and break down? Rather than revert?

Which proteins does it synthesize while in this "cyst" state? Are they any different from what they synthesize when they are not in this "cyst" state?

The spirochetes in this situation are not faced with the same issues in vivo. What it synthesizes and how it survives in the host is very different from what happens in a petri dish. What happens in its host?

This paper was written in 2000. I think some of these questions have been addressed in the past decade, though, haven't they?

Re: Info about Lyme round bodies (cysts)

Posted: Sun 1 Jan 2012 16:52
by Henry
ChuckG: May I say a few more things for clarification. In the presence of antibiotics, as well as under nutrient deprived conditions, bacteria -- including Borrelia-- are unable to make a component of their cell wall or membrane that is need for the bacterial cell to maintain its characteristic shape (a rod, a coccus, or a spirochete, as the case may be) and integrity. In the absence of this "rigidity" , the resulting bacterial cell is very much subject to the isotonic state of its surrounding medium. For example, in the absence of serum (protein) or high salt concentration, more water flows into the cell (by an effect very much like reverse osmosis) so that the cell begins then begins to swell -- and may even burst. So, the shape of the final product is not likely to be uniform in shape, but quite amorphous, depending upon the degree of residual rigidity left to maintain a semblance of the original cell structure or integrity. Consequently, all types of amorphous shapes might be present, as well as fragments of what once were single, whole bacterial cells; it would be ludicrous to try to categorize the different shapes found as though they were distinct entities or part of the natural growth cycle of Borrelia. If these changes were induced by antibiotics, additional antibiotics are not likely to be more effective in eliminating them since the resultant products are most likely at -- or very near-- an end stage. All that remains is for cells of the host immune defense system (e.g., macrophages) to do the final "mop up" and eliminate what's left. That will happen in time.

The real problem with calling these bodies "cysts" is that it create the false concept of Borrelia being encased in some sort of "cocoon-like" structure that protects them from inactivation by antibiotics or natural host defense mechanisms. Those who promote such a false image advocate the need to use "cyst busters" (e.g. flagyl and a variety of other unproven product) to get rid of these "cysts". There is no evidence that any of these "remedies" are the least bit beneficial; using them would be a absolute waste of money and could even result in great harm.

Re: Info about Lyme round bodies (cysts)

Posted: Sun 1 Jan 2012 17:12
by Henry
ChuckG: I forgot to comment on the issue of "reversibility" of these bodies -- that I will call sphaeroplasts for the sake of convenience-- to the original bacterial cell structure. There is a wealth of literature and research on this issue; all of it goes back to the 1950s-1960s when the mechanism of action of antibiotics was studied in great detail.

If bacteria are grown in the presence of antibiotics, sphaeroplasts will form as expected. However, if within a few hours after they are formed, the spheroplasts are placed in antibiotic free growth medium of suitable isotonicity, the intracellular antibiotic is "washed-out" of the cell and removed; this enables the bacterium then to make the cell wall or membrane component required for its characteristic shape and structure. It must be noted that reversibility is possible only within the first few hours after exposure to antibiotic; beyond that time, the effects are not reversible. Thus, if a patient is put on antibiotics at a dose sufficient to maintain a therapeutic concentration in the blood for an adequate period of time -- the details of course depend on the type of bacteria infection in question and the antibiotic used-- infection will be eliminated.

Re: Info about Lyme round bodies (cysts)

Posted: Sun 1 Jan 2012 21:56
by Camp Other
I think one of the important things to keep in mind is that while spirochetes are different from other bacteria, they aren't so different as to be completely alien. They have different survival strategies and a different sort of diderm from other gram-negative bacteria (it's really its own thing here, a genetically different category). But if one studies other bacteria, they will learn that there are similarities: Other bacteria can bleb. Other bacteria can form these spheroplasts and other forms when in adverse conditions. I think one odd experience I've had while reading through Lyme disease support web sites is that many people posting seem to think specific behaviors and structures are special to Borrelia burgdorferi and are things which set it apart - when in the microbio world, these behaviors and structures are common.

Henry said,
"Thus, if a patient is put on antibiotics at a dose sufficient to maintain a therapeutic concentration in the blood for an adequate period of time -- the details of course depend on the type of bacteria infection in question and the antibiotic used-- infection will be eliminated."
I think this has been one issue repeatedly brought up on various Lyme disease support groups: How does one know that the course of treatment they've been given is the correct therapeutic concentration to eliminate infection?

It's a good question, given that Borrelia tends to not want to live in the blood and heads towards tissues during dissemination. The concentration has to be high enough to hit it there. Also, the patient's immune status has to be pretty good or the baseline concentration may be inadequate for the job (and are we talking MIC or MBC here?).

I know patients who have had to have longer than typical antibiotic treatment for non-Borrelial infections because of their immune status (and also because they had a coinfection). So a longer treatment may need to be determined on an individual basis. It is also the case that studies referenced in the IDSA guidelines mention some treatment failures which happened with some patients - and in some cases, the patients were given additional treatment by the researchers which fell outside of the study scope. Some were cases of neuroborreliosis too, so the patients were given IV ceftriaxone. So really, to some degree there has to be some latitude in making the best therapeutic decision for the individual.

I think the one thing which I've found concerning in my research is that different strains have been shown to respond to different antibiotics quite differently - at least European strains. These are in vitro studies, though and I don't know how much follow-up there has been. It seems to me that if we had tests which determined right away which strains patients had that a more effective antibiotic could be given to them to begin with - much like how one tests for staph strains to find out what the best antibiotic is for treatment.

One confusing item for me is that I've read that Borrelia burgdorferi has not shown any signs of antibiotic resistance - or to be precise, I've read that other Lyme disease patients have said the IDSA has said Bb has not shown any signs of antibiotic resistance.

Antibiotic resistance in Bb has been investigated in vivo, in vitro, as well as in clinical reports. Bb is resistant to aminoglycosides, ciprofloxacin, rifampin, most first generation cephalosporins and some clinical isolates are erythromycin resistant. That last bit is important to know, because erythromycin has often been given to pregnant women instead of giving them doxycycline. (Barthold has also written about resistance in his own publications.) So I don't think this claim about what the IDSA said is correct... or if it is, then the IDSA must mean something else when they state Bb is not antibiotic resistant.