Research Funding Borrelia burgdorferi/and Miyamotoi ; Alz Plaques

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inmacdonald
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Research Funding Borrelia burgdorferi/and Miyamotoi ; Alz Plaques

Post by inmacdonald » Sun 16 Aug 2015 2:13

Hello to all on this forum: The following is the background for my ongoing Research into biofilms of Borrelia Burgdorferi
and biofilms of Borrelia Miyamotoi identified by Molecular Beacon DNA probe studies of Autopsy Alzheimer's Disease brain tissues:

I continue to work on my Borrelia/Alzheimer's Disease Research.
I commenced in year 2006 with my design of a molecular beacon DNA probe for Burgdorferi B31 Flagellin B- ( locus bbo0147). Beacons are special DNA probes and are a Patented Technology, licensed to users such as myself for Research Studies.

One use for Beacons is to deliver in real time the progress of
PCR amplification of a target sequence. Beacons are single stranded DNA ( maximum length 30 nucleotides) that are
forced into a circular configuration- the circle is "tied together"

..Gift wrapped if you will.,, by a "stem of nucleotides", and at the 5 ' end to one stem ssDNA is a fluorochrome( such as Fluorescein isethionate(FITC) -[Green fluorochrome]. At the 3' end of the other "arm" of the "Stem" nucleotides is a Quencher molecule. The Quencher molecular touches the Fluorescent molecule in the un-hybridized (circular) state.

When the Beacon Probe finds its target, It will hybridize with the Target nucleotide sequence ,,If and only IF ,, there is a 100%
complementary nucleotide sequence.
A SINGLE Nucleotide MISMATCH will prevent the Beacon from Hybridizing.
A stereo Conversion of a HYBRIDIZED MOLECULAR BEACON
will force the Nucleotide ( Fluorochrome + Quencher) to
"open up" and convert from a circular to a linear/planar configuration. The Hybridized Beacons are then free to release
a fluorescent light beam from the 5' end, because the
"open" planar Beacon has " defacto moved" the Quencher Molecule at the 3' end to such a great distance away from the
5' fluorochrome, that it no longer Quenches the Fluorescence.

For diagrams and line drawings of all of this verbiage,
please navigate to http://www.molecular-Beacons.com
Well, the above preamble is necessary to understand the significant of my Beacons for Borrelia Research Results.
In year 2006, I demonstrated that in a case of American
acquired Alzheimer's Disease, that my Molecular Beacon for
Fla B of ATCC Borrelia burgdorferi ( strain B31) (ss)
produced in each and every Alzheimer's plaque, and produced
fluorescence in the Granulo-Vacuolar Bodies which typify Alzheimer's disease. In year 2006, Dr. Eva Sapi and I commenced our research on Borrelia Biofilms [ In Vitro].Experience by me with the biofilms of borrelia, lead me immediately to document with photomicrographs in high resolution optics ( CytoViva) that Biofilms of Borrelia are often
Round and composed of various forms of Borrelia( mostly dot like granular borrelia, derived from Spiral Borrelia) . The biofilms are specialized COMMUNITIES of borrelia - encircled and protected by a glue like Matrix of DNA, RNa, and Proteins, derived from once living but now Dead Borrelia. Water channels traverse the biofilm communities- providing a canal system for delivery of nutrients and for Waste removal from the community.

I was, as a pathologist, very well aware of the properties of the Plaques of Alzheimer's Disease. They are usually round, and come in small diameter, medium diameter, and large diameter sizes. Traversing the Alzheimer's Plaques, are a canal like system of optically empty spaces forming a network entirely identical to the Water Channel system of Biofilm Communities.In year 2006, I made the Hypothesis that Alzheimer's plaques
are actually biofilm communities of Borrelia [ and possibly other infection agents]. I recalled that my Molecular Beacons had clearly and sharply labeled the Alzheimer's plaques, and that with my Beacons I could identify a system of black empty spaces forming a network of channels inside of the Alzheimer's Plaque.

I wrote my idea that Biofilms of Borrelia are the Cause of Alzheimer's Plaques into my lectures from that moment forward. Today, with a second set of Molecular Beacons - specific for Miyamotoi borrelia Flagellin B and glpQ DNA, I have discovered that the Alzheimer's Plaques are also formed by the DNA of Miyamotoi borrelia.

Further, I have proven with my In Situ DNa Hybridization with My Miyamotoi and Burgdorferi Molecular beacons, that
Chimeras of Borrelia [ incorporating the DNa of Burgdorferi and the DNA of Miyamotoi inside of a single borrelia spirochete] are present in the Alzheimer's brain at autopsy, and that Chimeras of borrelia Miyamotoi/burgdorferi are present inside of the biofilm communities (Alzheimer's Plaques) of Alzheimer's disease.

Now recall that DNA directs the synthesis of all proteins.
For a Pure Burgdorferi borrelia, a family of proteins is produced,
and for a pure mutated variant of burgdorferi, a family of proteins is produced. For a pure Miyamotoi borrelia, a different family of proteins is produced.

Vaccines are manufactured by purifying a very few proteins from an infectome, and adding an adjuvant, to make the body produce antibodies to the proteins in the Vaccine.
What does my discovery of Chimeric (Burgdorferi/Miyamotoi)
mean for The dream of developing a Vaccine to protect
mankind against All Borrelia infections.?
Chimeras of Borrelia - sound the DEATH KNELL to
all possibility for an effective Vaccine for All infections Borrelia.

I am recruiting donors to send small amounts of money to support my DNA Probe Molecular Beacons Research.
If many people with Lyme and other borrelia infections
each send in a small donation, the impact will be a success
in raising the dollars to develop New Beacon DNA Probes
for Lusitania, Valisania, Bissetti, Kurtenbachii, Americanum, Andersoni, and Sinica, and many more yet to be named borrelia infections which produce negative or inconclusive results in current European blood testing and in American Blood Testing.

Each New Beacon DNA Probe costs $1000. to manufacture, purify, Validate, and quality control. I dream of the day when 20 or more Borrelia strain Molecular Beacon DNA probes will be available for Direct DNA based detection of borrelia spirochetein Blood and in spinal fluid, and in solid tissue, and in seminal fluid, and in amniotic fluid.
In that time, soon to arrive, Diagnosis by DNA based technology will replace Blood testing....for Antibodies alone,...

All of my Research has ALWAYS Been FREE to All- and in the public domain. I have never Patented any of my discoveries. I never will patent any of my work. I have never charged any patient for my work on their behalf with my research studies, and I never will.

I hope that you will share this letter, with your colleagues, and with Lyme/Borreliosis Patients in France and in other European Theater countries.

My Research Funding website is the Link Below:

http://www.gofundme.com/z3v2a2k

I have more images to share with you from my work in
discovery of Borrelia spirochetes with my Beacon DNa probes in
Multiple Sclerosis Spinal fluid specimens, and in Alzheimer's Disease Autopsy Brains.

Respectfully,
Alan B MacDonald MD,FCAP
August 15, 2015

User avatar
inmacdonald
Posts: 977
Joined: Fri 13 Jan 2012 22:32

Re: Research Funding Borrelia burgdorferi/and Miyamotoi ; Alz Plaques

Post by inmacdonald » Sun 16 Aug 2015 2:37

Images from Alzheimer's Brain: with Molecular Beacons DNA PROBES FOR BURGDORFERI DNA AND MIYAMOTOI DNA:

BIOFILM COMMUNITIES OF BORRELIA MIYAMOTOI (RED COLOR) AND BURGDORFERI BORRELIA ( GREEN COLOR)
WITH MOLECUALR BEACONS ( DNA PROBES ABSOLUTELY SPECIFIC AT 100% MATCH FOR HYBRIDIZATION LEVEL)

------------------BURGDORFERI DNA PROBE (MOLECULAR BEACONS) FLUORESCENCE IN SITU DNA HYBRIDIZATION [FISH METHOD]-----{Alzheimer's AUTOPSY BRAIN TISSUE -- HIPPOCAMPUS ---WHITE/GREEN COLOR = DNA OF BURGDORFERI BORRELIA- GREEN BACKGROUND = THIOFLAVIN T STAINING OF AMYLOID PLAQUES IN ALZHEIMER'S DISEASE
Biofilms of Burgdorferi Borreli Inside of Amyloid Plaques of Alzheimer's Disease.png
BURGDORFERI borrelia DNA probe outlines biofilms of burgdorferi INSIDE of Alzheimer's Amyloid Plaques- IN ALZHEIMER'S DISEASE- ARE BIOFILMS OF BORRELIA IN GRANULAR FORM SURROUNDED BY DNA OF BORRELIA MIYAMOTOI IN EXTRACELLUALR MATRIX ENVELOPEMENT OF THE BIOFILM COMMUNITY WHICH IS DERIVED FROM DNA OF ONCE LIVING BUT NOW DEAD BORRELIA MIYAMOTOI SPIROCHETES ( ecm= EXTRACELLULAR MATRIX MATERIAL}
Biofilms of Burgdorferi Borreli Inside of Amyloid Plaques of Alzheimer's Disease.png (362.29 KiB) Viewed 1454 times

_______________________________________________________________________________MIYAMOTOI DNA PROBE (MOLECULAR BEACONS) [FISH METHOD] ---ALZHEIMER'S DISEASE AUTOPSY BRAIN TISSUE --- WHITE/RED COLOR = DNA OF MIYAMOTOI BORRELIA - RED BACKGROUND IS CONGO RED STAIN TO SHOW AMYLOID IN AMYLOID PLAQUES OF ALZHEIMER'S DISEASE PLAQUES
Miyamotoi Borrelia Biofilm community inside of Amyloid Alzheimer's disease Plaque.png
Miyamotoi Borrelia Biofilm community inside of Amyloid Alzheimer's disease Plaque.png (192.8 KiB) Viewed 1454 times

LINK TO RESEARCH FUNDING WEBSITE FOR ALAN B. MACDONALD, MD

http://www.gofundme.com/z3v2a2k

Thank you for your support of this research!

RESPECTFULLY SUBMITTED,
ALAN B. MACDONALD, MD , FELLOW, COLLEGE OF AMERICAN PATHOLOGISTS
AUGUST 15,2015

Joanne60
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Joined: Mon 13 Feb 2012 15:49
Location: Guildford Surrey UK

Re: Research Funding Borrelia burgdorferi/and Miyamotoi ; Alz Plaques

Post by Joanne60 » Sun 16 Aug 2015 12:53

Dr MacDonald thank you for posting this which I have shared elsewhere, one comment and question from a Facebook friend that maybe you wouldn't mind answering please.
Only just occurred to me that the idea of the molecular beacons is very similar to that of monoclonal antibodies which people may be more familiar with - at least a generation of A level biology students is! We presume that European strains have the same behaviour re Alzheimer's plaques as miyamotoi and burgdorferi? Is there any reason why Alan does not mention garinii and afzelii in his ambitions for beacon probes?

Lorima
Posts: 914
Joined: Mon 29 Oct 2007 20:47

Re: Research Funding Borrelia burgdorferi/and Miyamotoi ; Alz Plaques

Post by Lorima » Sun 16 Aug 2015 16:00

This is work that needs to be done. It may be the most important work that needs to be done right now.
It checks out for me with regard to general biological plausibility, and the right methodology for the job.

Count me in.
"I have to understand the world, you see."
Richard Feynman

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inmacdonald
Posts: 977
Joined: Fri 13 Jan 2012 22:32

Re: Research Funding Borrelia burgdorferi/and Miyamotoi ; Alz Plaques

Post by inmacdonald » Tue 25 Aug 2015 14:25

Dear Lorima,

Yesterday I completed a draft of the exact method for the imaging of
Amyloid Alzheimer plaques with double Label technique as follows: first
perform Fluorescence in situ DNA Hybridization on Alzheimer's autopsy
Hippocampus tissue glass slides using the Molecular Beacon DNA Probe for
Miyamotoi Borrelia Flagellin B (FISH method). Second stain the completed
FISH slides with an amyloid stain - I used Congo Red stain under polarized
white light for one set of slides and I used Thioflavin T stain for Amyloid
for a second batch of slides.

I then captured photomicrographs of Amyoid Alzheimer's plaques at high
magnification in sharp image resolution . For each amyloid plaque I
immediately , without changing the focus or the field of view, removed the
light source for Amyloid photo capture and substituted Epifluorescence with
the monochromatic light for excitation of the Miyamotoi Borrelia Beacon DNA
probe ( 640 mm ) and photographed the identical field of view . This
captured the Borrelia DNA which had hybridized to the Beacon , ( Cy5 Fluor.
Label). In each case studied, every Amyloid plaque contained a biofilm of
Borrelia Miyamotoi inside of the Topography of the plaque. Two techniques
for Amyloid staining ( Congo Red in polarized white light. And Thioflavin T
fluorescence at 500 mm light) produced indentical results. A Borrelia
Biofilm was dwelling INSIDE of each and every Amyloid plaque.
Under the email address which your friend provided to me, I will,send to
you the 17 page document. You will receive it by the Hightail email
service.
I will send to you, privately, a 17 page compendium , which I compiled last night,
demonstrating pairs of images (A) Amyloid Stained Plaques (B) DNA probe (FISH)
hybridized images for Miyamotoi borrelia Flagellin B DNA.mRNA.- in the identical
microscopic field of view.
The Biofilms of borrelia Dwell INSIDE of Amyloid Alzheimer's Plaques.
Some of my Images in the 17 page Compendium have previously been POSTED
on my Research Funding Website:
http://www.gofundme.comz3v2a2k.
Thank you very much for your support and very substantial encouragement.

Sincerely,
Alan
August 25,2015

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