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#4854 - 02/17/10 12:30 AM
Conversion to CWD and Cystic form questions
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Tighidden
New Researcher
Registered: 02/17/10
Posts: 1
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In your book The Top Ten Lyme Disease Treatments on page 83., you say that protein synthesis inhibitors stop the bacteria from being able to shift from form to form, which makes sense. I was just wondering if you had more info on how it stops it from doing this or if there are any articles or material you could point me in the direction of that would give me more information on this. The main reason I ask is because I thought that i have seen studies done in vitro with Doxy that showed the spirochete transforming to the cystic form, I'll have to look again for those. But, in the mean time if you wouldn't mind shedding some light on this for me it would be much appreciated. Thanks.
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#4855 - 02/17/10 03:31 AM
Re: Conversion to CWD and Cystic form questions
[Re: Tighidden]
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BryanRosner
Forum Veteran
Registered: 11/08/06
Posts: 263
Loc: Mountains of Northern CA
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Here is an excerpt from my book Lyme Disease and Rife Machines . There is also some new evidence since I wrote that book, I did a blog post on it, but I can't seem to find the post. This video may also interest you.
PRIMARY SCIENTIFIC STUDIES CITED IN THIS CHAPTER
Appendix G contains endnotes that cite research sources throughout the book. However, the following scientific studies were important enough to be included in the text of this book.
Induction of cystic forms by different stress conditions in Borrelia burgdorferi. Murgia R, Cinco M.
Spirochete Laboratory, Dipartimento di Scienze Biomediche, Universita' degli Studi di Trieste, Via Fleming 22, 34127 Trieste, Italy.
Cystic forms of Borrelia burgdorferi might represent a low metabolic activity state or phase of B. burgdorferi cells that allows the spirochete to survive in a hostile environment until conditions are favourable to multiply again. In this study we evaluated the rate of cyst formation induced by oxidative stress, pH variations, and heating, reconversion of cysts to vegetative forms, and some aspects of their metabolic activity. We observed cyst formation in the presence of extreme pH values, and at high temperature, but the best production of cystic forms was observed in the presence of H2O2. When transferred to BSK II medium, the cystic forms reconverted to spirochetes in relation to their age and type of induction treatment. Furthermore, we demonstrated a low metabolic activity of cystic forms by measuring amino acid incorporation. Overall, these data suggest that the phenomenon of conversion to cysts by B. burgdorferi provides a limited survival potential. This short-term survival, however, gives borreliae an additional chance to overcome unfavourable environmental conditions.
PMID: 14961976 [PubMed - in process]
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Conversion of Borrelia garinii cystic forms to motile spirochetes in vivo. Gruntar I, Malovrh T, Murgia R, Cinco M.
Institute of Microbiology and Parasitology, Veterinary Faculty, Ljubljana, Slovenia. gruntaig@mail.vf.uni-lj.si
Cystic forms (also called spheroplasts or starvation forms) and their ability to reconvert into normal motile spirochetes have already been demonstrated in the Borrelia burgdorferi sensu lato complex. The aim of this study was to determine whether motile B. garinii could develop from cystic forms, not only in vitro but also in vivo, in cyst-inoculated mice. The cysts prepared in distilled water were able to reconvert into normal motile spirochetes at any time during in vitro experiments, lasting one month, even after freeze-thawing of the cysts. Motile spirochetes were successfully isolated from 2 out of 15 mice inoculated intraperitoneally with cystic forms, showing the infectivity of the cysts. The demonstrated capacity of the cysts to reconvert into motile spirochetes in vivo and their surprising resistance to adverse environmental conditions should lead to further studies on the role and function of these forms in Lyme disease.
PMID: 11478686 [PubMed - indexed for MEDLINE]
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Serum-starvation-induced changes in protein synthesis and morph
ology of Borrelia burgdorferi. Alban PS, Johnson PW, Nelson DR.
Department of Biochemistry, Microbiology, and Molecular Genetics, University of Rhode Island, Kingston 02881, USA.
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]
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A rapid method for generating cystic forms of Borrelia burgdorferi, and their reversal to mobile spirochetes. Brorson O, Brorson SH.
Department of Microbiology, Vestfold Sentralsykehus, Tonsberg, Norway.
Mobile Borrelia burgdorferi were transferred to distilled water (10(6) per ml). The cultures were observed by dark field microscopy (DFM), interference contrast microscopy (ICM) and transmission electron microscopy (TEM). 95% of the spirochetes were converted to cysts after 1 min, and after 4 h no normal mobile borreliae were observed. When transferred to growth medium (BSK-H), the cysts became smaller and more irregular, and were filled with organic substances. After 1 day, 1-5 thin structures sprouted from the cysts. They continued to grow in both length and thickness until they attained a normal spirochetal structure. Finally, these new-born spirochetes detached from the cysts, by which time their mobility had become normal. The present method for producing large amounts of cystic forms of B. burgdorferi is well suited for further studies of this unique microbe.
PMID: 10052721 [PubMed - indexed for MEDLINE]
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Effects of penicillin, ceftriaxone, and doxycycline on morphology of Borrelia burgdorferi. Kersten A, Poitschek C, Rauch S, Aberer E.
Department of Dermatology, University of Vienna, Austria.
Antibiotic therapy with penicillin, doxycycline, and ceftriaxone has proven to be effective for the treatment of Lyme borreliosis. In some patients, however, it was noticed that borreliae can survival in the tissues in spite of seemingly adequate therapy. For a better understanding of this phenomenon, we investigated the different modes of degeneration of Borrelia burgdorferi suspensions during a 96-h exposure to various antibiotics. By dark-field microscopy and ultrastructural investigations, increasing blebbing and the gradual formation of granular and cystic structures could be followed during the exposure time. Although antibiotic concentrations at the MIC at which 90% of organisms are inhibited after 72 h were 80% or even greater, motile organisms were still present after incubation with penicillin and doxycycline but not after incubation with ceftriaxone. By transmission electron microscopy, intact spirochetal parts, mostly situated in cysts, were seen up to 96 h after exposure with all three antibiotics tested. According to experiences from studies with other spirochetes it is suggested that encysted borreliae, granules, and the remaining blebs might be responsible for the ongoing antigenic stimulus leading to complaints of chronic Lyme borreliosis.
PMID: 7625800 [PubMed - indexed for MEDLINE]
In vitro activity of vancomycin against the spirochete Borrelia burgdorferi. Dever LL; Jorgensen
Antimicrobial Agents & Chemotherapy, 37:1115-21.
JH; Barbour AG.
"Approximately 75% of cells exposed to either penicillin or vancomycin had one or more large membrane blebs, designated gemmas (6), whereas untreated B31 cells in log-phase growth had only occasional (<20% of cells) small blebs that were smaller than those seen in treated cells... Thin sections of B31 cells treated with penicillin or vancomycin were indistinguishable from one another. Both demonstrated numerous gemmas... Numerous smaller spherical blebs were associated with the outer membrane of treated cells and were also found separate from the cell membranes. Untreated cells demonstrated only occasional smaller spherical blebs, found in association with and separate from the outer membrane. Rod-shaped forms or extremely long spirochetes were not observed in treated cultures."
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Action of penicillin on Borrelia hermsii. Barbour AG; Todd
1982 Antimicrobial Agents & Chemotherapy, 21:823-9.
WJ; Stoenner
Benzylpenicillin at its minimum bactericidal concentration induced formation of large spherical structures. These structures were bounded by one or both cellular membranes and, in some thin sections, appeared to contain material from disrupted protoplasmic cylinders... they are consistent in appearance with spheroplasts (20,29). ...A prominent electron microscopic finding was the abundance of small membranous blebs or vesicles in the penicillin-treated culture. Blebbing of the outer membrane is said to occur when spirochetes are under 'adverse conditions.'
_________________________
Bryan Rosner
DISCLAIMER: Bryan Rosner is a journalist, not a healthcare provider or doctor.
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