PCR results are in

I have sent the specimen being documented in this blog, for mtDNA analysis, and the preliminary results are in. It is unknown at this time. They ran queries for fungal sequences and it came up negative. Many, who are familiar with microbiology have suggested it may be fungal in origin. This would be natural assumption, judging by morphology, as seen in micrograph above.
It is apicomplexan, but classification may be a task ahead. The DNA is still available for further sequencing, which may have to be done.
Is it novel? Well, it certainly has not been well documented. Since people like myself are infected with this or variations of this, as a human disease I would have expected a faster hit.

Documenting curious microbe. Pathogen?

Click on image for closer scrutinity


I have isolated a very interesting microbe, and have photographed it through high performance camera at special settings under dark field at 600+-x.
The microbe is motile, and tends to roll. It is made of smaller subunits, which I never would have seen without the computer images. The subunits are green, with red on polar end. I have observed the units break off and migrate away from mother unit. The unit, which I call "sponge integer" has been cultivated from sample placed in 30% bleach with purified sterile water and incubated at 37+ degrees C. The sample was blood obtained from microscopic slide wet mount.
The donor, has multiple acquired symptomologies without definitive diagnosis. Suspicion of bartonella has been noted by specialist, but I do not see any indications of this.

If you study the image, you can see the surrounding subunits, these are found throughout sample. There are also other ameoboid type microbes in the solution, and scattered biofilms.
I speculate the red terminals are acidic molecular residues, and the green is neutral PH. This type of configuration is noted in microbes I have observed with intracellular parasites. I am not jumping to conclusions here, but feel these microbes are worthy of recording for future reference.

More on parasites

I have been trying to upload some important clips, but they would take.

Two cells at high magnification

The first photo is from my "oocysts". The fact these are occysts will be confirmed soon through mtDNA analysis. It is not fungal [unless recombinant], because sporozoits were grown and observed, and even infected other cells---check old posts for documentation on that. Sorry, the photo was taken by commercial camera, but it is high magnification. It is at same setting as second micrograph shown.


This second micrograph is a very high magnification of 2 cells from on going culture of unknown spores. There has been much activity between metabolites, microbes etc., in solution, and the appearance of cells which I say could be oocysts. But yeast cells, and penicillium molds have also been isolated from original source in dry plate culture. I have compared the spores from the {obtained} fungal specimens to objects in free solution broth. From observation, I have eliminated those as being the source of growing bodies in solution, which I believe are parasites.

The cells in micrograph look close to cells I have isolated from my own body, but are from individual hundreds of miles away, whom I have never even met.

Immature microbes, becoming something weird

Click on image for close up
The first micrograph is taken under normal light setting, shows flat view of microbe. [BTW this microbe glides]

The second is under dark field, the light refracts, so there is limited clarity. The colors are very close, though. There is methylene blue in the solution, which causes some of this effect. The energy of the microbe, does emit color. It was seen moving, before capture. They do not live long under glass.

The yeast in question

This was the yeast grown directly from slide sample, although I isolated 3 strains of penicillum as well. The yeast was pure white.
The cells making up the penicillium had some similarities with the growing cells [in pic 3 posts down from here] but the ones in question, seem to be different.
I would have guessed yeast right off the bat, but the microbe in question seems to be directly associated with these cells. I am almost sure they are sporozoites, and if this thought is not swayed, I will suggest to donor, that we have it analyzed genetically.

Check out the suspect oocysts in previous photo's and take note of the cell wall. It has definite thickness--it is a different cell.

More micrographs

These 3 micrographs are as follows: 1) bio film seen as typical film in sample [stained with methylene blue] it appears almost 3D under direct observation, with the stained section in top layer 2) typical bio film, with heavy globulins 3)oocysts[?] which are under investigation--stained with MB. It is not surprising the suspect oocysts stained heavy. This was seen in oocysts from my personal specimen sample obtained from diarrhea contained entirely in mucus.
These oocysts[?] developed from blood specimen of person with chronic symptomology.

Preliminary micrograph of suspect microbe

Click on image for size increase. [4-5 um dark field]

This is just a quick peek at what I "suspect" is a participant in the process which results in the emergence of oocysts. This one is later stage, and there are small proteins tethered to one end.
In the host, the first observation from blood, looked like a rod bacteria. In later experiments, this same ron type bacteria became present. Other forms resembling this form, had proteins which moved in and conjugated with this structure. It becomes covered over time, and starts to produce globulin structures. This is a far as I get.
There are also cysts in same solution, so to much going on for me to make any kind of conclusions.

Click on image for larger picture.

This micrograph is from a sample which contained strange, non-bacterial microbe, which shows signs of complex organization. I have gone through great lengths to promote differentiation of microbe into other stages of development. This compares to trying to make a caterpiller make a chrysalis, and become a butterfly!
Being lead by intuition, some scientific method, and experience, I eventually came up with these structures, which I think are the oocysts made by unknown microbes.
Any comments from experienced microbiologists will be most welcome.

This is a common protozoa to "my" gut. It is big, by comparison to other protozoa and produces cysts. It is hard to tell if these are problematic, because I have only seen attack by leukocytes one time.
The apicomplexan parasites collected previously, have been isolated in mucus, therefore have been associated with immune response, and deemed problematic.
Futhermore, they have been examined in culture for a variety of phases, and been evaluated.

Protozoa

 

This is a protozoa, common to my gut. There seems to be some activity, around the microbe. When the pseudopods extend, it looks like tiny cysts are being secreted. Unsure of that part.
The micrograph was taken by my new camera, and adjusted under different lighting than original view. The detail came out nicely. Click on image for enhanced view.
The background particles are common bacteria.

stuff on proteasome from science mag

Ubiquitin, Proteasomes, and the Aging Brain

Douglas A. Gray, Maria Tsirigotis, and John Woulfe

Douglas A. Gray is at the Ottawa Regional Cancer Centre, Ottawa, Ontario, Canada K1H 1C4 and the Ottawa Health Research Institute, Ottawa, Ontario, Canada K1Y 4E9. Maria Tsirigotis is at the Ottawa Regional Cancer Centre, Ottawa, Ontario, Canada K1H 1C4. John Woulfe is at the Ottawa Health Research Institute, Ottawa, Ontario, Canada K1Y 4E9 and in the Department of Pathology, Ottawa Hospital, Ottawa, Canada K1Y 4E9. E-mail: Doug.Gray@orcc.on.ca
http://sageke.sciencemag.org/cgi/content/full/sageke;2003/34/re6

Key Words: ubiquitin • proteasome • neurodegeneration

Abstract: Ubiquitinated proteinaceous inclusions are the hallmark of many neurodegenerative diseases. Inefficient proteolysis might lead to the accumulation and ultimate deposition of potentially toxic entities as inclusions within neurons or glial cells. This hypothesis is supported by genetic evidence both from patient populations and from engineered mutations in genes that encode ubiquitin/proteasome components in mice. The appearance of similar inclusions in the brains of elderly individuals of normal and subclinical conditions begs the question of whether there is a general age-related decline in the ability of the ubiquitin/proteasome pathway (UPP) to recognize and eliminate abnormal proteins, and whether such a decline would be reflected by changes in the abundance or activity of some or all components of the UPP. Here we describe alterations in the aging mammalian brain that correlate with a decline in the function of the UPP and review the evidence for age-related changes in specific UPP components. These alterations are discussed within the context of prevalent theories of aging.

Citation: D. A. Gray, M. Tsirigotis, J. Woulfe, Ubiquitin, Proteasomes, and the Aging Brain. Sci. SAGE KE 2003

I am leading up to something here, but my time is being sucked into a black-hole somehow. It is these processes I will try and show, that cause neurological problems with those who have stealth parasites\pathogens.