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wrotek
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Posted: Thu Mar 2nd, 2006 21:37 |
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Lyme movie, the following was posted on Eurolyme yahoo group:
Whist we are on the subject of darkfield lyme movies.. Here is one of my
own.
This was done from a drop of my blood this afternoon and video taken about
an hour afterwards.
Warning - not for the feint hearted!
http://www.qualityelectronicdesigns.com/lyme_movie.zip
Best,
J.
It is terrifying for me that u can culture this things only from one drop of blood. Some one will use Your shaving machine by mistake and He is doomed.
wrotek
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Lyme reflux chronic pain fatigue depression 125D36 Ph1Sep05 Ph2Oct06 Ph3Apr07 homebound in low lux NoIRs 25D<7 Oct06
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Dr Trevor Marshall
Research Team
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Posted: Thu Mar 2nd, 2006 22:49 |
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This seems to be an authentic movie, using a variant of Dr Andy Wright's methods, and the poor guy/gal who gave the blood is certainly quite ill.
I am also interested to know what equipment was used to generate this image - it clearly was not the Bradford microscope that Andy uses. It would be nice to come up with some low cost equipment that we could proliferate around the world to do this imaging.
I would point out, however, that a physician member of ILADS has told me that the bacterial tubules coming out of the dead cell are just artifacts, fragments of membrane, and not bacterial L-forms. I don't agree with this, of course, but am trying to point out that even this level of evidence is not persuasive for some folks.
..Trevor..
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jsandman001
Member in Phase 2
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Posted: Thu Mar 2nd, 2006 23:30 |
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Dr. Marshall,
I have been doing these samples for a while on myself and a few others. I talked at great length with Dr. Andy Wright and Dr. Mattman at the Chicago conference to learn how to make a vaseline prep slide and still show them some of my movies. Didnt know there was interest but since there is i will send you some and tell me what you think. I even took blood from my ear and like Dr. Mattman said at the conference .........definately more aggressive movement. There is no secret here folks just old school techniques. You can find a used microscope on ebay with a darkfield condenser and some high powered objectives. My setup also contains a phase contrast condenser. Some high powered objectives and some phase optics. A doctor can be in business for $600.00-800.00 looking at eveyones blood although it is best to start looking at the blood 12 hours after making the slide then view it on up to 36 hours........you will be absolutely amazed at what you find..you will see bacteria hatching out of the red blood cells.........you can actually see in phase contrast what looks to be millions of bacteria moving around in white blood cells. The picture you showed of the different stages of growth Dr. Marshall are all apparent if you spend time under the scope to look. Fron the little horseshoes to the ones that look like dumbells to amazingly long chain of pearls. Anyone who is interested in seeing a few of my little movies just mail me at jamesanders01@comcast.net and for the people who could tell there doctors about it .......the old American Optical model 110 is hard to beat as it is still used at most of the labs i have been to and can be had for a song on ebay.
Thank You,
James Sanders
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CFS, Lyme, Colitis,25D=12,6/30/2008, MP1/07, MPh2 10/07, restart Ph1 01/08, klonipin, Zyrtec, Asacol, NoIRs, cover up, low lux home
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Dr Trevor Marshall
Research Team
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Posted: Thu Mar 2nd, 2006 23:46 |
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James,
Yes, a few months ago I was looking at some of the Olympus models on Ebay. But what condenser and objective lens magnification and video camera accessory did you use? Andy was saying that he needed x8000, but it seems to me that was being achieved on the Bradford by something akin to "digital zoom" techniques. So I wasn't too keen to use a low resolution camera adapter for the microscope. What did you use for the camera objective?
It seems that Scenalyzer capture software would do an excellent job on the time-lapse. What did you use for that?
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Alayne
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Posted: Thu Mar 2nd, 2006 23:52 |
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Wrotek, am I correct in understanding this movie is of your own blood? 
This is most fascinating and frightening to see. -Alayne
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ME/CFS/FM 6/05:25D-34 1,25D-69, 11/07:25D-8 1,25-37, Sick 6-11 mos/yr x30+yrs. NoIRs/Avoid Sun/D/Use Zinc oxide. 11/17/05-Ph1, 5/06-MPh2, 12/06-MPh2#2, 6/07-MPh2#3,1/08-Ph2, 4/08-Ph3, NonMP Meds: MThistle/Calc&Mag/Lysine
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wrotek
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Posted: Thu Mar 2nd, 2006 23:53 |
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Dr Marshall i will track as much informations as i will be able to, this looks like very good magnification, red cells are very big. I was interested in microscopy and also wanted to buy one and meybe i will do  Dark field FUNCTION is not so expensive like phase contrast do, but good CCD camera i very expensive. jsandman001 i would love to see Yours movie also.
____________________
Lyme reflux chronic pain fatigue depression 125D36 Ph1Sep05 Ph2Oct06 Ph3Apr07 homebound in low lux NoIRs 25D<7 Oct06
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wrotek
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Posted: Thu Mar 2nd, 2006 23:55 |
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No Alayne it is not my , it is a guy called Jason from Eurolyme yahoo group. I will try to contact him
The Olympus models are fascinating, the olympus website about microscopy knowledge also remarkable. Moving flash animations explain everything. http://www.olympusmicro.com/
When i was talking about microscopes months ago, someone told me(he knew it from school from a teacher) that Bradford Variable microscope is just normal microscope but it has bigger field of view and from magnification definition it make bradford magnifying not 1600x but 8000x and more. Physics? Last edited on Fri Mar 3rd, 2006 01:37 by wrotek
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Lyme reflux chronic pain fatigue depression 125D36 Ph1Sep05 Ph2Oct06 Ph3Apr07 homebound in low lux NoIRs 25D<7 Oct06
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jsandman001
Member in Phase 2
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Posted: Fri Mar 3rd, 2006 02:11 |
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Hello All,
Dr. Marshall wish i would have known you were looking for an Olympus as i just sold a Vanox AH2 ......same as Dr. Mattman used i believe. It was a monster at 115lbs. I would have donated it to the Foundation. Equipped with darkfield, phase,DIC .....i actually got it to use the Direct Interference on the bacteria thinking the light shearing might show more detail but was wrong it actually just washed the bacteria out. Yeah labs are dumping nice Olympus BH2's now so you can get them cheap.
Dr. Marshall and everyone interested. I have worked on a Bradford and what you are seeing above 1200x to 1400x is empty magnification......i know you hear 15,000 to 30,000x resolution but Ernst Abbe in the 1800's found out that a transmitted light microscope can only resolve to about 1400x. Empty magnification for those interested is like taking your 4x6 picture to the local convienient store and making an 8x10 out of it.....sure you can blow it up but the resolution stays the same. So you end up with an 8x10 looking not as clear as the 4x6.
Just to show you now i have a Leitz Ortholux made in the 1950's ....very good research stand. Dr. Marshall was showing slides and Dr. Matmann had some slides of people doing this in the 1800's but unfortunately darkfield microscopy found its way into alternative medicine and when that happened people discounted that and pleomorphism as a fraud.........Thankfully Dr. Marshall and the few others are picking up on the work of the old school crowd when doctors actually had microscopes in there offices and did some of their own pathology.
Dr. Wright will tell you and i can tell you the bacteria show up very nice for observation at about 1000x. I have several 100x objectives along with 10x eyepieces and 12.5x eyepieces. You must make a vaseline prep slide which is taking a very tiny drop of blood. Like the size of the back of a stick pin. Take a q-tip and put vaseline on the edge of the coverslip then put it on an angle on the drop of blood on the slide.....so you dont look at a lot of air.
Then oil the condenser to the slide then oil the objective to the slide for the highest magnification you can achieve...and after 10 hours you will get it .....but once you get it its very simple to repeat ....i mailed Dr. Wright forever trying to get it right.
I started out just using my point and shoot digital camera (an Olympus Camedia 2.1 megapixel that plays small movies) It does 4x zoom i think so you would be looking at 4000x magnification. I now have 2 Dage ccd cameras that do 1200 lines per inch of resolution but have to mount one to my trinocular port. They are $5000.00 cameras that i picked up on ebay for between $40.00 and $60.00.
Microscopes are on the rise now as everyone wants to get into cloning but deal are still to be had.
For stills which i have a good cannon digital now you can use a stacking program to get really nice images........time lapse not sure about decent software but why do that when you could get a sony ccd and adapter for a few hundred and run it out in real time to a monitor and vhs or dvd recorder to send out.
Wrotek got your mail i will send you a few movies out after i send to Dr. Marshall. Red blood cells are 6 to 8 micron wait till you see this 50 to 60 micron snake running around in my blood .......definately not artifacts or fragments of a membrane....but i will let you decide
Thanks........now back to my herx and a upset 5 year old.
James Sanders
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CFS, Lyme, Colitis,25D=12,6/30/2008, MP1/07, MPh2 10/07, restart Ph1 01/08, klonipin, Zyrtec, Asacol, NoIRs, cover up, low lux home
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John McDonald
Foundation Director
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Posted: Fri Mar 3rd, 2006 06:32 |
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The Bradford scope just has to be half shamanism. I design and make microscopes for the semiconductor industry and I teach a seminar on microscopy once a year at a conference of failure analysis engineers. Nearly anything that could be done with a visible microscope was done or described by Ernst Abbe quite a long time ago. We use darkfield quite a bit but only as an alternate contrast enhancement technique. For darkfield, the light is introduced outside the normal field of the objective and will not go into the objective unless deflected, hence the field is "dark". If something deflects the light into the field it appears bright, against the black field. In brightfield the light is introduced right through the objective and you rely on the specimen to deflect the light out of the field, so it appears dark on a bright field. Well, heck, you can make a dark field image by shining a gooseneck lamp on the outside of the objective. It isn't as elegant as an expensive darkfield objective, but it does produce a usable darkfield image. You can also use a ringlight mounted on the end of the objective. There is a firm in Simi Valley that can make a custom ring-light for you for a few hundred dollars. If you can see these microbes in darkfield, then there are probably a few brightfield methods that will also show them.
The absolute theoretical limit for resolution for a white light microscope, with a good lens, oil immersion and carefull setup, is 0.2 microns. You can just match that to young eye with 600x magnification. Anything above about 1000x is empty magnification, no additional resolution but the same blur reproduced in larger size.
There are some fabulous animated java microscope tutorials at http://www.molecularexpressions.com.
____________________
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jdc
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Posted: Fri Mar 3rd, 2006 08:49 |
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Trevor,
Can you or someone else provide the upshot to this technical discussion?
1. Using darkfield microscopy is it likely that a person can view the spirochete infections inside microphages of one's own blood?
2. Is it the spirochetes that might be viewed or only the bacteria when they have taken a walled form and are in the bloodstream?
3. Is the viewing of intracellular microbes only possible on some occasions during infection?
4. Would the viewing be useful at all for disease monitoring--for example, assessing the no. of spirochetes or their characteristics over time?
I've got a pretty good feeling my doctor would be interested in microscopic viewing if you think it would be worthwhile. I also think that it would help him get over a bit of doubt about the infectious etiology of th1 diseases.
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PA 6/2000; started protocol 8/2004 Vit D 40/22; ACE 70;Minocin, 100 Mg.; hiatus for surgery from 3/17--4/17/06; 5/1/2006 restarted phase one, retesting of vit D 25: D
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Jvancan
Member in Phase 3
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Posted: Fri Mar 3rd, 2006 08:57 |
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Very interesting topic! Maybe we could make a special topic on microscopy and the MP? With subjects as wich magnification, equipment, slides preparing etc.
@John:
The absolute theoretical limit for resolution for a white light microscope, with a good lens, oil immersion and carefull setup, is 0.2 microns. You can just match that to young eye with 600x magnification. Anything above about 1000x is empty magnification, no additional resolution but the same blur reproduced in larger size.
But how does A. Wright gets this high magnification of 8000x? Is this done by digital magnification? But if I understood it wright at 1000x you should also be able to see spiros, with the wright illumination and slide preparing?
JeroenLast edited on Fri Mar 3rd, 2006 09:03 by Jvancan
____________________
Lyme 125D69 25D32 Ph1May 06 Ph2July06 Ph3Dec06 break Mar07 to Jan08 25D12.6Jan08 NoIRs occ lite exp Ph1Jan08 Ph2Apr08
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Dr Trevor Marshall
Research Team
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Posted: Fri Mar 3rd, 2006 11:28 |
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Jeroen,
Alan Cantwell used Oil Immersion to get 1000x. That could be the lack of "young eye" John mentioned, however. Andy uses a commercial 'Bradford' microscope at his local University, which, as the posts above describe, is at least partly smoke and mirrors. I saw a low-cost dark-field microscopy set put together by Dr Jamie Deckoff-Jones in 2004, and she was getting excellent videos. I recall she was using x400.
Another 'pioneer' in this microscopy, Dr Marie Kroun (in Denmark) has been developing the use of low-cost computer microscopes (like the one at this URL http://tinyurl.com/fry5x ). Personally, I have no problem with using a low-cost DV camera with lens attachment, as I think the video processing (time-lapse, etc) would be significantly eased by using DV, with concurrent still-capture as well.
I have already proposed to the FDA that monitoring of blood by this technique (which I will call "Andy's technique," for simplicity) should form part of the endpoints for the stage 3 clinical trial we have been planning, and for which we applied for designation last September. I was basing that on the ease with which Dr Deckoff-Jones had been viewing the pathogens, but eventually the rubber has to hit the road and we have to start some real imaging ourselves, I guess. That is where everybody's experience, especially the data from James and John, becomes invaluable. Thank you all
ps: there is one big caveat on all this - such a microscope would almost certainly come under the FDA regulations as a "diagnostic" device if any claims are made for its efficacy in diagnosis. I have enough red-tape right now, and this really precludes its widespread use by physicians as part of their practice, except "to satisfy their idle curiosity" (that's a legal term).
pps: if you don't know what we are talking about in this thread, take a look at the DVD of Dr Andy Wright's presentation in the Plenary session at Chicago. And Dr Lida Mattman's, too.
You may purchase DVDs of the Chicago Conference at the AutoImmunityResearch.org Most of the background material explaining the pathogenesis, and the curative therapy for these Th1 diseases, is covered during the 12 hours of presentations, panels and tutorials in that collection.
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Dr Trevor Marshall
Research Team
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Posted: Fri Mar 3rd, 2006 11:34 |
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Oh - Dr Bela Bozsik in Hungary has developed a flourescent antibody (similar to that used by Dr Whittaker at Bowen) to detect the presence of Borrelia, and reports that it is present in the bloodwork he has seen. Andy Wright reports one of twelve CFS patients whose blood-artifacts were not Borrelia, however. Which tends to confirm my "multiple species" and "horizontal transfer of DNA" presumptions.
We need some flourescent antibodies developed for other probable pathogens. Is there anybody developing these antibodies besides Dr Bozsik, Dr Whittaker and Dr Jardin (in South Africa)? At "30th Lyme" Dr Whittaker did talk about an E.coli specific antibody which she generated decades ago as part of her PhD Thesis research.
DVDs of the 30TH Anniversary of Lyme Disease - An Educational Medical Conference in Farmington, CT on May 7, 2005 (including Dr. Marshall's presentation) may be ordered at:
http://www.ctlymedisease.org/order.htm
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Jvancan
Member in Phase 3
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Posted: Fri Mar 3rd, 2006 12:47 |
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Dr. Marshall,
There is a Lab in the Netherlands, wich also uses a Fluorescence test for Borrelia, it's really new. They use the same technique wich the Dutch goverment uses for Legionella Fluorsence tests in the water supply. I have an intermediate picture from them of my own blood wich show spirochetes. But the photo is blurry, they have some problems with non lyced cells.
As far as I know theire test is accredittet for spirochete forms. But they know there are more forms then only spirochetes.
P.S. Trevor I saw the Bresser usb camera. I have purchased a Bresser Researcher Microscope, also vissible at that site (this one: http://tinyurl.com/mf8u5). It has the abbility to use darkfield and phase-contrast condersors. I hope I can see some forms with this device. So im really interesting in this topic.Last edited on Fri Mar 3rd, 2006 12:51 by Jvancan
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Lyme 125D69 25D32 Ph1May 06 Ph2July06 Ph3Dec06 break Mar07 to Jan08 25D12.6Jan08 NoIRs occ lite exp Ph1Jan08 Ph2Apr08
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LeAnne
Member in Phase 3
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Posted: Fri Mar 3rd, 2006 16:12 |
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Thank you wrotek for this thread. This is the specific type of information that I am looking for to give people who ask me how exactly these bacteria are detected.
LeAnne
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Neuro-Sarcoidosis/lungs, spleen, nervous system, skin lesions, 125D66, MP 8/05, Ph1 3/06, Ph3 7/06, NoIRs, low lux home, cover up, 25D9 Sep07
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jrfoutin
Research Team
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Posted: Fri Mar 3rd, 2006 16:37 |
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All,
Do look at the still images of coccoid, rod, "ghost" and giant bodies (Russell's) shown in Alan Cantwell's papers on JOIMR. Most are x1000 acid fast in oil.
His latest papers (reviews of literature) at JOIMR discuss infectious bacteria in both Hodgkins and Scleroderma. He provides excellent still images (usually about mid way in article).
LINK TO JOIMR HODGKINS
LINK TO JOIMR SCLERODERMA
Both articles call for science to explore beyond rather standardized microsope techniques. Cantwell's Scleroderma article discussion of H. Pylori relevance to today's science:
"What was essential in the discovery of stomach Helicobacter was the use of a special tissue stain to properly identify these bacteria in ulcer tissue."
Cantwell's Hodgkin's article identifies tool and tissue sources:
"...use of the ordinary light microscope, using the oil immersion lens at a magnification of 1000 times. Microphotographs of bacteria in tissue sections of the heart, lung, lymph nodes, skin, pancreas, cerebrum, bone and marrow..."
...as well as Cantwell's perspective about the element of time:
"cultures should be examined and re-examined over a period of time, rather than being discarded after several days, as is the case with most 'routine' bacterial cultures."
In fact, most of Cantwell's article titles and abstracts (he is acutely aware of the busy scientist's "skim" reading process) attend to not only the first level recognition of these bacteria but the next step of looking into the pleomorphic nature of these bacteria and a subtle call to action for further research on discovering their changling nature and the life cycle of these little beasties.
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My personal feelings? Those of you with the toys and inclination, PLEASE read up, collaborate, use scrupulous unimpeachable processes, take notes/photos/movies and go for it!
____________________
Sarcoidosis 125D61, MP10/05 ModP2 12/05 Ph2 6/06 Ph3 10/06, NoIRs limited outings covered, 2/08 25D6.2, 10/08 25D6.9
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wrotek
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Posted: Fri Mar 3rd, 2006 16:51 |
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Jason equipment is just normal BIO microscope, with 5 watt LED bought on ebay.
This is an interesting patented DUaldur technique, makes everything hold still except living moving spirochetes.
http://lymerick.ulmarweb.dk/Sheffield2005/Bozsik/Dualdur.wmv
http://lymerick.ulmarweb.dk/videomicroscopy.htm
Meybe for ILADS and for others this could solve problems with differentiating between moving spirochetees and not moving cell wall filament components sticking out from the cells.
The visible wave lenghts are between violet 400nm (0,4microns), 0,450 microns (blue) and 700nm red (0.7 microns), so i guess the best resolution is to place violet iris to use the shortest wave lenght beeing carefull cause ultraviolet kill pathogens right ?
0.2 microns will be blurred as i presume
Ivancan do u have the dark field to bresser ? or phase contrast ? I was thinking about buying bresser with dark field once a time.
And other interesting thing, if Yours tests are negative just buy microscope and make a movie with yours spirochetes and u will have evidence "that it is not in Your head".
But some people will always find an answere, this are not spiros because sth end sth
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Lyme reflux chronic pain fatigue depression 125D36 Ph1Sep05 Ph2Oct06 Ph3Apr07 homebound in low lux NoIRs 25D<7 Oct06
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John McDonald
Foundation Director
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Posted: Fri Mar 3rd, 2006 17:22 |
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I am happy to help if I can. I am a numbers guy but I get in the lab when I can. I can help find a particular condensor or a phase contrast lens and I can design and assemble a microscope from specs. However I work in the physical sciences and though I have used biological instruments, I have only 'tourist' experience with florescence, florescent tags, biological samples and the like. I can also help with various contrast mechanisms and I know how to tease the best possible resolution from a given scope. I also know the various microscope vendors. Off the top of my head, a good multi-purpose research grade microscope, nicely fitted out, and new out of the box, might cost about $15,000. A single high quality objective runs several thousand dollars. Price is where Jame's skills may be invaluable.
Another good bet might be to vist CalTech or UCLA. Some of those labs have some mighty nice equipment and if we only need a few hours here and there, I bet we could arrange a visit. Even Pepperdine or Cal Lutheran might have suitable gear.
James seems to have pieced together an excellent instrument but Chicago is a long way from the Los Angeles area. Can you show us a photo of your setup?
For years I felt that a physicist had a big disadvantage in a barter economy. What commonly useful thing do I know? Eventually a psychologist friend encouraged me. He said that I can always fill black holes in driveways.
john
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paulalbert
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Posted: Fri Mar 3rd, 2006 17:27 |
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How much can the pervasiveness of a given L-form type infection be revealed from a single snapshot taken from a high-powered microscope? It has been my understanding that everyone has these infections in some shape or form, so it would not be enough to merely find one or two. Could the presence of just a couple nanobacteria be a sufficient indication that there is some form of TH1 illlness at work?
Also, is it conceivable that as imaging techniques for these bugs becomes more sophisticated that some kind of standardized measure of the pervasiveness of an infection might emerge? If so, could such a measure one day be economically and practically feasible for use in diagnosing patients? Could such a measure still have meaning even in light of the fact that infections are confined to certain tissues?
I guess I don't need to say it, but this types of measure (in addition to the D ratio, which is only of value before treatment begins) would really be helpful in assigning a degree to which an illness has advanced as well as identifying various cases of previously subclinical infection. This hypothetical measure could show, to a higher degree of certainty, how a person is responding to the Protocol, or any external stimuli, for that matter.
I'm sure there are a number of reasons why this measure does not now exist including the fact that it's rather novel, but exactly why not and if it is conceivably possible are still lost on me.
PaulLast edited on Fri Mar 3rd, 2006 17:55 by paulalbert
____________________
Diag CFS 6.03 / sympt since 9.02 / exercise, food intol, sleep prob / 1,25D: 16, 4.06; 1,25D:27, 25D:26 7.04; 1,25D:43, 25D:6 6.05; 1,25D:17, 25D:8 8.05; / MP: 7.04 / Ph. 3 / Bacteriality
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Jvancan
Member in Phase 3
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Posted: Fri Mar 3rd, 2006 18:11 |
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Paul,
How much can the pervasiveness of a given L-form type infection be revealed from a single snapshot taken from a high-powered microscope? It has been my understanding that everyone has these infections in some shape or form, so it would not be enough to merely find one or two. Could the presence of just a couple nanobacteria be a sufficient indication that there is some form of TH1 illlness at work?
Shouldn't blood be sterile according to mainstream medicine? Even a few forms are no good IMO, but not every one with a few forms has "yet" a Th1 disease. I think when you have these forms, with symptoms of a Th1-disease and/or a D-ratio disturbance, then I think these snapshots are a good evidence IMO.
Jeroen
____________________
Lyme 125D69 25D32 Ph1May 06 Ph2July06 Ph3Dec06 break Mar07 to Jan08 25D12.6Jan08 NoIRs occ lite exp Ph1Jan08 Ph2Apr08
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