RELATED/OVERLAPPING INNOVATIONS IN HEALTH/ENERGY/TRANSPORT/FARMING AND INFRASTRUCTURE

INNOVATIONS CONNEXES OU CHEVAUCHANT LES DOMAINES DE LA SANTE, DE L'ENERGIE, DES TRANSPORTS, DE L'AGRICULTURE ET DES INFRASTRUCTURES

Claims

Claims

Chitosan/chitin-calcium phosphate composites/nacre/mollusk shell/shells from
lobsters/crawdads/crayfish...polysaccharide...also aragonite, calcite and
vaterite.

Any and all Uses for chitosan's antibacterial properties and its low toxicity
for human and
creatures ingestion.

We can use it as a spray (eg. mix with drinks in hepatitis prevalent
countries), flavoured
and/or unflavoured (no taste) - an additive to any and all foods mixtures
and/or beverages,
as well as spray on cup (prevention e.coli, salmonella, avian flu) spray a
film on masks for
any and/or all diseases that are airborne, including diseases that are spread
by fluid transfer.
We could also inject this substance via empty T-cells (and other immune cells
eg.
macrophages, phagocytes and lymph) that bind to HIV AIDS, filling these immune
cells
whose membrane proteins bind with HIV AIDS cells and deliver chitosan (same as
cancer
and any and all diseases whose infection method can be identified to bind with
the immune
cells membranes between proteins and receptors to deliver chitosan to kill the
diseases and
then we could repopulate the disease battle zone with stem cells (possibly use
HOX genes
and/or building blocks of HOX) and differentiated stem cells appropriate to
the battle zone.
The delivery of these concoctions can be via multi micro syringes to ensure
neither too much
and/or too little of the concoctions to the map of the infected battle zone.
We can use the above for any and all uses to kill off bacteria and/or viruses.
(Jz 0.7%)

We could use the chitosan in conjunction with zeolite, semi-permeable
membrane, and test
for heavy metals - if there is high amount of heavy metals in a batch we could
plasma torch
the batch to burn for energy.

We could use it for sewage/tea manure and/or all organic wastes. We could also
use it to
caked/coated onto a multi level horizontal/vertical matrix/scaffold to soak in
reservoir tanks,
until the water has reached safe levels.

We could use it to develop fluoride and/or chlorine and/or paste.

We could develop jugs where caked on perforated blocks drip off and mix with
the chitosan
cleansing the water. (Tea Bags - 25% GP), Powder and/or dissolving pill.

Also a large diameter straw with permeable caked/coated on dyed (to indicate
level filtration
ability remaining) chitosan. And even polluted rivers and/or lakes and/or
wells and/or
cisterns...whereby a large amount of chitosan is slowly discharged (and since
chitosan is a
renewable resource, we can mass produce the substance.

We could dye all the chitosan to observe the remaining concentrations.



(DLD Asia 40%)
Also to add to polluted water sources.
Also Salt Water to Energy and Fresh Water Technologies.
We could also rear any and/or all chitosan sources such as crayfish/crawdads,
in multiple
levels of tanks in huge underground bunkers, possibly growing algae and other
biocrops
and/or any and/or all energy sources eg. solar, wind mills, TALL GARVITY TO
ELECTRICITY INVENTION...

We could also build aquariums (tanks/reservoirs) with a sunken floor to drain
through pipe
that is covered by a rubber stopper (like toilet stoppers), so that by lifting
the stopper
(flushing action) the stopper lid opens to the spout/pipe.

The spout/pipe immediately under the stopper has a grill/sift with small
enough holes to avoid
the fish/crawdads...falling through.

We can use a squeegee/rake to sweep the uneaten food and/or feces, down the
spout/pipe,
where these material is drained away.

We could add calcium and/or chitin, to the water when the creatures are
moulting.

We could try to clone chitosan the stem cells and differentiate into the layer
cells (precusor
cells to the chitosan shell (GP 1%)) under moulting that produce chitosan
exoskeleton.
And/or produce molecules that have the same basic structure as chitosan.

Chitin (C8H13O5N)n is a long-chain polymer of a N-acetylglucosamine.

Chitosan can also be used as scaffolding for stem cell, and differentiated
planted artificially
grown organs.

Chitosan could also be distributed in powdered and/or sprayed form to stop red
tide.
Chitosan used for any and all detergents and/or oil/tar sands bitumen
separator, (tailings
ponds GP 13.5%), any and all industrial uses especially food and beverage
industries, wet
and dry tissue and toilet paper, (paper towel 13.5% GP), film lined any and
all bags - paper
to garbage bags. Pulp and Paper (eg. card boxes) made with less need for
poisonous
preservatives. We could use the chitosan for (refining any and all petroleum
and bio fuels
GP 10%).

Chitosan might also be used as a spray on preservatives such as wood, any and
all spray on
applications (2.5%).

Chitosan could also be used as soaps for animals. Chitosan could also be added
to
(compost?), and/or an additives wet vacuums, and/or wet air filters (with
removable
cartridges Jeff 1%), it could also be caked on/coated on any and all seeds,
especially bio
crops, and/or chitosan could be used at the end of the need for micro
organisms, as a way of
treating the micro organisms out the fermentation process to kill off
potential microbial
threats.



Chitosan could be used in powdered form to treat foot odour, dry and remove
odour from
shoes and/boots that are wet inside (ski/snowboard boots), kitchen clothing,
fabric softener,
carpet powder, spray/powder for any and all home uses on furniture/fruit
bowl...any and/or
all hospital uses, and office, and leisure (eg. where hepatitis, dengue fever,
hemorrhagic
fever ...
On another note, we plan to genetically bioengineer potatoes, and yams, and
sweet
potatoes, and taro roots to third world countries, where rice paddies costs
are going up, and
rice farming polluting the water supply (runoff from fertilizer and
pesticides) and rice farming
carries waterborne diseases such as malaria. Furthermore potatoes are probably
more
productive in terms of bulk value) as well potato farming is the USA is
suffering from low
cost, meaning it is a cheaper source of food for developing nations struggling
to put food on
the table.

We could use chitosan to refine any and all oil and gases

We could also create cardboard boxes and biodegradable bags and wrapping -
which can
be made in different sizes and durability with a paper/cardboard box/bag and
wrapping
maker that is like a printer controlled by the computers, especially where the
material
contained is perishable foods, (eg. inner lining of candies, and snack foods).

We could make chitosan tooth paste and mouth wash, and/or gum and/or candy
mints for
fresh breath.

We could use chitosan and its properties to make a healthier palm oil and any
and all oil
(cooking products GP 0.1%).

We could use chitosan to clean up any and all hazardous materials. And use to
clean up
contaminated lands; land fills, slow dissolving slow delivery to aquaculture
pens possibly
hoses with sprinkling.

Hoses/trickling/sprinkling (eg. capillaries that reach the width/length of
shellfish commercial
farms that release) the chitosan in agriculture as well as farming feed.

As a an additional farming note we plan grow mass quantities of potatoes in
developing
countries where land is cheap, and rice prices are rising, while potato
profits in decreasing in
America are lowering. Potatoes are possibly more productive in terms of
filling the stomach
with less land. Potatoes use less water loss to evaporation. Rice fields can
cause water
borne diseases (including parasites and malaria). We could build green houses
whose roofs
are shaped like a spout/funnel that drains rain into a tank. We could also use
Gerard Voon's
salt to fresh water and energy invention to provide water. Local sources of
water could be
filtered with chitosan.

We are considering growing hydropohnically grown potatoes plants in floating
trays. We will
deliver into the water and/or substrate chitosan to prevent rot. We might
recover the
chitosan via, membrane, filter and/or drying. We can grow craysish/crawdads
under the
floating platforms, the walls need to be higher and transparent to keep the
crayfish/crawdads
from escaping, yet not interefere with light on the potato plants above. We
could use tea
manure for the fertilization of tanks, and for germinating tanks, no
fertilizer, where the
crayfish/crawdads are safest.

(GP 1%)



We might also grow flats of algae (of optimal depths) with chitosan mixing in
tea manure to
harvest bio fuel.

If we make the floating platform tall enough to support the neck we could grow
corn and/or
any and all bio crops and high profit crops (eg. ice wine). We could even
incorporate these
hydroponic platforms into multiple level glass sky scrapper buildings. We
could rotate these
flats of floating platforms in circles (GP 1% solely) to get better sun
exposure - more even or
slow/stop/reverse to give certain regions more light. We could put in the
centre of the
building a Gerard Voon's patented TALL GRAVITY TO ELECTRICITY INVENTION
(and/or
any and all building such as offices/condos/hotels/resorts/apartments at its
centre GP 5%)
and industrial uses for the centre as well and top levels buildings of
underground shopping
malls) whereby any and all crops are on multiple level floating platform flats
that rotate
around the buildings. We could also incorporate windmills on top or sides of
the building.

We could rotate the entire platforms deck/level spinning around to get the
most of the sun
relative to the different needs of the platforms.

We could also build these spinning farms on platforms in the sea/oceans
including polluted
places.

We could also grow jacobra in hot dry climates by using low water required
substrates such
as lava rock and foam.

We could even use lattice skeletal structure of the building that are open
walls, in climates
that are non-extreme with the transparent roofs that funnel/spout the water
down to a
tank/reservoir.

We could use the centre/core of the building to chum the corn and microbes to
(churn)
ferment and produce ethanol. We plan to use sound and magnetics in pulses to
stimulate/induce the micro organisms to (possibly imitating pulsating sounds
that cause
optimal metabolism and temperature and timing possibly in cycles so the micro
organisms
are pushed in the extreme one way at the cost/price of their long term health)
to grow,
proliferate and produce enzymes that ferment the corn.

We could use plasma torch/laser to burn any and all wastes, such as coal
effluent, pulp and
paper discharge (where the liquid discharge can be bound to chitosan and then
dried and
burnt with plasma torch/laser), any and all hazardous waste and all
industrial/raw and all
along the development stream of wastes produced (GP 1%) including recycling.

We could tie-in developers to be green (recycling) by identifying any and all
parts taking
apart and all buildings (sky/scrappers, houses, condominiums...) before
demolition and/or
renovation and refurbish their
appliances/computers/televisions/audio/video/electronics
furniture and the building materials like bricks separated from the dry wall
and wooden
doors/frames, walls... Some of the material can be refurbished (eg. computer
with CD - for
education, Internet (to learn about anything and everything from the world's
knowledge) and
email (to make friends even business connections worldwide) and Word
capabilities (to be
able to communicate and express in the Arts, Business, Science... papers of
your ideas and
understanding, perhaps internet web page designing program) and supplied to
the poor
worldwide.




We could also try to increase proliferation of stem cells by administering the
ambilify drug,
viagra and/or cialis...

We could have any and all plants uptake chitosan to fight fungi, rot, disease,
toxins and
heavy metals and produce long shelf life in fruits and vegetables and their by
products.
We plan to take the homeodomain protein (genes) WUSHEL (WUS), LEAFY (LFY), and

AGAMOUS (AG) to cultivate and grow stem cells of (high value plants - biofuel
crops,
ginseng, coffee, tea, rare plants).We are also testing GARP and bZIP. And
explore
phytohormone cytokinin, transcriptional network and factors and how these
processes
regulate proliferation.

Growth hormone cytokinin is at most effective when WUSCHEL stops the ARR genes

feedback loop.

Ethylene is a major factor that drives the plant cells to proliferate.

We also plan to use epigenetic mechanisms which have had some success in non-
plant
stem cell proliferation.

The purpose of cloning is to try to grow valuable fruits and vegetables by,
1.) tissue culture of
the stem cells and then differentiating them into desired cells (fruits and/or
vegetables,
kernels), 2.) multiplication effect from less starter cells needed to grow
next generation's crop
size (eg. rather than planting kernel by kemel, one kemel's many stem cells
can be used to
produce multiplications of starter plants for next generation crops), 3.)
hybridization, isolating
stem cells from the most productive and best characteristics for suited
purposes of such
crops.

We could also grow potatoes in our towers (glass sky scrappers), our plan is
to submerge
the potatoes und a platform with a hole so the stem can grow above, and we add
chitosan in
the medium below to prevent fungi and rot.

We plan to use pressure and temperature - to manure/sewage/any and all organic
waste
/tea manure... (possibly a vacuum surrounding the targeted raw materials) to
create nitrogen,
that can be used as fertilizer.

We can use the following for nitrogen fixation, including legume family
Fabaceae:
Family. Genera

Betulaceae (Birch): Alnus (Alder)
Casuarinaceae (she-oaks):
Allocasuarina
Casuarina
Gymnostoma
Coriariaceae: Coriaria
Datiscaceae: Datisca
Elaeagnaceae (oleaster):




Elaeagnus (silverberry)
Hippophae (sea-buckthorn)
Shepherdia (buffaloberries)
Myricaceae:
Morella arborea
Myrica
Comptonia
Rhamnaceae (buckthorn):
Ceanothus
Colletia
Discaria
Kentrothamnus
Retanilla
Trevoa
Rosaceae (rose):
Cercocarpus (mountain mahogany)
Chamaebatia (mountain misery)
Purshia (bitterbrush or cliff-rose)
Dryas

(Taken from Wikipedia) There are also several nitrogen-fixing symbiotic
associations that
involve cyanobacteria (such as Nostoc). These include some lichens such as
Lobaria and
Peltiqera:

.cndot. Mosquito fern (Azolla species)
.cndot. Cycads
.cndot. Gunnera
Microorganisms that fix nitrogen
.cndot. Diazotrophs
.cndot. Cyanobacteria
.cndot. Azotobacteraceae
.cndot. Rhizobia
.cndot. Frankia

We could also develop microbes (and/or any and all enzymes) to combat bad
breath,
microbes for fermentation (gasses and/or sugars/carbohydrates/cellulose to
ethanol and/or
compost...any/and all waste organic (eg sewage; manure) and any and all -
mixed with
temperature (mirrors cheaper and plasma torch augmentation - eg. if ore heat
is needed
and/or days that are not sunny) and heat for oil/tar sands - to eliminate
difficult to process
waste, seriously unwanted and of little recycling value we could plasma torch
it (with Gerard
Voon's TALL GARVITY TO ELECTRICITY INVENTION V - generating the energy to
power
the plasma torch).

Microbial growth requires cell signalling when there is a quorum (to grow
large colonies of
microbes one needs to start with a grouping of the microbes), and biofilms
that appear at the
same time are an indiicator if not a cause if not and an affect. In humans, in
the case of
infections, bio films can block the immune cells from reacing the
perpertrating microbes.




Perhaps by adding Dermatix or other scar removing substances, the patient
could have their
biofilm removed, and then stem cells and localized differentiated cells
(probably from the
stem cell stock) can be injected possibly with chitosan, to repopulate the and
clean up the
infection site.

We could use genetic circuits on any and all genes of any and all creatures.
There are four
main chemicals; A for adenine; C for cytosine; G guanine; T thymine.

We could use DNA synthesizers.

Using mixed chemicals (solvents) that form molecules ... that create short
strings to short
strings to longer chains, and joined into doubel stranded DNA and copied.

In addition we could use furanone from a long-leaf sea weed to prevent the
microbes from
building biofilm. This substance can be used for out breaks of antibiotic
resistant infections,
including digestive and any and all illnesses or unwanted presence of
biofilms, for human
and any and all creatures, applications, mix with stem cells, localized
differentiated cells,
chitosan Even cancer, possibly inflamatory over active immune systems...(or
under active
immune systems).

Nitrogen Fixation by Cyanobacteria

Cyanobacteria inhabit nearly all illuminated environments on Earth and play
key roles in the
carbon and nitrogen cycle of the biosphere. Generally, cyanobacteria are able
to utilize a
variety of inorganic and organic sources of combined nitrogen, like nitrate,
nitrite, ammonium,
urea or some amino acids. Several cyanobacterial strains are also capable of
diazotrophic
growth. Genome sequencing has provided a large amount of information on the
genetic
basis of nitrogen metabolism and its control in different cyanobacteria.
Comparative
genomics, together with functional studies, has led to a significant advance
in this field over
the past years. 2-oxoglutarate has turned out to be the central signalling
molecule reflecting
the carbon/nitrogen balance of cyanobacteria. Central players of nitrogen
control are the
global transcriptional factor NtcA, which controls the expression of many
genes involved in
nitrogen metabolism, as well as the P II signalling protein, which fine-tunes
cellular activities in
response to changing C/N conditions. These two proteins are sensors of the
cellular 2-
oxoglutarate level and have been conserved in all cyanobacteria. In contrast,
the adaptation
to nitrogen starvation involves heterogeneous responses in different strains



Nitrogen fixations can be used to extract nitrogen from sewage/manure/any and
all organic
wastes to have a purer/ cleaner fertilizer, that can as a final step be mixed
with chitosan to
eliminate microbes. We could synthesize the genes (and/or clone the entire
microbe)
responsibly for most effective nitrogen fixing as well ethanol from cellulose
as well a s bio
crops... We could also use the gene synthesis technique, to create new life of
any and all
forms, differentiate stem cell pools into required specialized cells. And/or
reprogram old cells
to revert to precursor and/or stem cells (eg. HOX, Homeodomain cells) perhaps
replenish the
telomerase. We could examine the tole of telomerase with and without endings
to see if the
reason a stem cells stops proliferating is directly caused by telomerase
shortening or if its
shortening with age is only another age related coincidence.

For Inflamatory diseases (antagonists) formyl peptide receptor like 1 receptor
(FPRL
1),whereby CCR1 ligand and CKB8-1 recruits monocytes and neutrophils and FRPL1
CKB8
attracts monocytes, dendritic cells, and resting lymphocytes. Mix the CCR1 and
FRPL1 and
CKB8-1 to recruit/attract immune cells to the infected sites.

One application is to take a large population of stem cells and differentiate
them (or tissue
culture) them into White cells, T-cells, macrophages, phagocytosis, (possibly)
B-cells,
cytotoxic, and remove the contents from these cells and refill them with
(electro shock, laser
fuse and/or micro syringe) material that would be toxic fluid and/or salts
and/or chitosan that
enter the virus AIDS/HIV virus until the virus bursts...

The AIDS/HIV has proteins, sugar-contains theglycoprotein gp 120 on its
envelope,
"recognizes" a protein on helper T-cells, macrophages, phagocytosis,
(possibly) B-cells,
cytotoxic named CD4, and physically associates with it. The CD4 [Cluster of
Differentiation
Antigen No. 4] protein is a normal part of a helper (both Th1 and Th2) T-
cell's membrane.
Thus, CD4 is a specific receptor for HIV.

Besides AIDS, formyl peptide receptor like 1 receptor (FPRL 1),whereby CCR1
ligand and
CKB8-1 recruits can be used with stem cell line grown immune cells (of variety
and amount
most effective for the procedure) and micro inject into any and all
bacteria/virus infected
regions of the body (to recruit/atract immune cells to), as well we could put
chitosan into the
mix to help kill microbes). We need to study the genes that activate and
deactivate
inflammation to treat diseases that are inflammatory related.

Diseases that can be treated include, but are not limited to, peripheral -
chronic inflammation-
related diseases, for example: chronic inflammation; thrombosis;
atherosclerosis; restenosis;
chronic venous insufficiency; recurrent bacterial infections; sepsis;
cutaneous infections;
renal disease; glomerulonephritis; fibrotic lung disease; allergic disease;
IBS; rheumatorid
arthritis and acute bronchiolitis. Central nervous system-macroglia and
microglia related
diseases, for example: neurodegenerative diseases; Alzheimer's disease;
Multiple sclerosis;
Parkinson's disease; neuroinflammation; HIV-associated neurological diseases;
HIV-associated
dementia; CNS bacterial infections; brain Toxoplasma gondii; Acanthamoeba
infections; Listeria infections; prion diseases; subacute spongiform
encephalopathies and
macular degeneration may also be treated.
We plan to test the chemokine receptors eg. CCR1, CCR2, CCR5, CCR8, CXR2 and
CXCR4 as well as cytokines to act as targeting mechanisms for immune cells.



We plan to try either or both Dermatix or scar removing products or furanone
to try to remove
the plaque/fiber tangles from Alzheimer's patients, then repopolate the area
with 1. stem
cells, 2. neutrophins, 3. localized differentiated cell types, 4. stimulus
pulsating electrode, all
these techniques are also applicable for Mad Cow Disease, Parkinsons Disease -
acetylcholine, norepinephrine, serotonin, soma-tostatin as well as melatonin,
these
treatments can also be used for patients with spinal cord illnesses treating
their neurons and
dendrites.


Gene therapies can be transfection, viral infection, microinjection, or fusion
of vesciles.

Proteins including anti-inflammatory (to treat inflamatory related illinesses)
ctyokines, growth
factors/antioxidants protect neurons when present in the extracellular space
(the presence of
a screctory signal sequence) the protein protects when inside a neuron - eg.
protein is an
antioxidant that localizes to an intracellular organelle. Neuron uptakes of
these proteins are
known as protein tranduction domain. Further conversion of stem cell into
neurons
include...nerve growth factor (NGF), glial-derived neutrophic factor (GDNF),
hepatocyte
growth factor (HGF), brain-derived neutrotrophic factor (BDNF), ciliary
neutrotrophic factor
(CNTF), fibrobalst growth factor 2 (FGF-2), neutrophin 3 (NT 3) and
transforming growth
factor B (TGF-B). We are using these factors to enhance proliferation and
cloning (including
stem cells).


We plan to use binding proteins calbindin D28K, to proliferate any and/all of
our cell cultures
(especially stem cells and cloning) and all treatments from excitotoxins, pro-
oxidants, serum
withdrawl, also from B-amyloid, excitotoxins, hypogiycaemia, cynaide, as well
as helping
metabolism, ATP concentrations and mitochondrial potential.

The inhibitor of apoptosis proteins (IAPs) are a family of antiapoptotic
proteins that bind and
inhibit caspases 3, 7, and/or 9. We plan to use IAP's to proliferate any and
all cells including
stem cells, differentiated cells of any kind, including immune cells for
fighting diseases, brain
damage...The IAP proteins contain a BIR (baculovirus IAP repeat) a domain next
to the
amino-terminus. The BIR domain can bind some caspases. A large proportion of
the IAP
family of proteins block proteolytic activation of caspase-3 and -7. XIAP,
cIAP-1 and cIAP-2
appear to block cytochrome c-induced activation of caspase-9, thereby
preventing initiation
of the caspase cascade. IAP-1 and cIAP-2 were first identified as components
in the
cytosolic death domain-induced complex associated with the TNF family of
receptors, they
may inhibit apoptosis by additional mechanisms. We plan to use these
mechanisms to
proliferate cells (especially stem cells and cloning).


Other antiatpotopic, via cdk inhibitor include flavonoid derative that
inhibits cdk 1, 2 and 4.
Olomoucine and roscovitine which are purine derivatives that inhibit cdk1, 2
and 5 and also
early response kinase 1 and/or MAP kinase activities. These substances are
effective in
saving and perhaps increasing proliferation (eg. stem cells or at least CGN
and cloning) of
trophic factor deprivation and DNA damage. Particularly induced by KCL
withdrawl.


FLIP/FLAME is highly homologous to caspase-8. It does not, however, contain
the active site
required for proteolytic activity. FLIP appears to compete with caspase-8 for
binding to the
cytosolic receptor complex, thereby preventing the activation of the caspase
cascade in
response to members of the TNF family of ligands. The exact in vivo influence
of the IAP
family of proteins on apoptosis. Using this mechanism we plan to augment
proliferation of
cells (especially stem cells and cloning).




The IE1 and IE2 proteins each inhibit the induction of apoptosis by tumor
necrosis factor
alpha or by the E1B 19-kDa-protein-deficient adenovirus but not by irradiation
with UV light.
We plan to use IE1 and IE2 proteins to proliferate any and all cells including
stem cells and
cloning, differentiated cells of any kind, including immune cells for fighting
diseases, brain
damage...


The 14-3-3 protein is also associated to proliferate cells, eg. stem cells and
cloning,
differentiated cells, immune cells, bone, neurons...


We are also using Heat Shock Protein 27 and 72 and 70 and 54 to make cells
(eg. stem cells
and cloning) proliferate.


We are also using BAG-4, BAG proteins, EBV-EBER, Imp-1, ki-67, bcl-2,
survivin, Bax, fas,
c-myc, and p53 to make cells (eg. stem cells and cloning) proliferate.


In mammal cells, survivin colocalizes with the mitotic apparatus including B
tubulin,
microtubules, centrosomes, and kinetochores. Inhibition of survivin with anti-
survivin antibody
results in delayed metaphase and produces mitotic cells with shorter and less
dense mitotic
spindles to be used for cell culture (stem cells and cloning).


For females we might try to obtain more eggs from females by managing the c-
jun protein, c-
fos protein, estrogen receptors alpha and beta (ER-.alpha. and ER-.beta.),
progesterone receptor
(PR), and Ki-67for their roles in proliferation (including stem cells and
cloning)and apoptosis
of glandular epithelial cells.


We are also using III collagen (eg. coated dishes), together with ascorbic cid
and ascorbic 2-
phosphate and long effecting vitamin C derivative and also epidermal growth
factor (to
further proliferate cells culture (especially stem cells and cloning).


We are also using Polycomb Group gene complexes (PcG) to proliferate cell
culture (eg.
stem cells and cloning), once we want to use the stem cells and cloning for
patient treatment
(eg. body parts), we can silence the PcG by using multiprotein complex PRC1,
PRC2 and
PhoRC...


We are studying using amino acids and glucose and water...


We also plan to develop microbes (eg. pre/pro-biotics) that act on the mouth
breath and
flatulence to remove the bad odours (perhaps mixed with zeolite - eg. powdered
and/or pre-
mixed with liquids).


Amino acid classes Class Amino acids hydrophobic Norleucine, Met, Ala, Val,
Leu, Ile
neutral hydrophilic Cys, Ser, Thr acidic Asp, Glu basic Asn, Gln, His, Lys,
Arg disrupt chain
conformation Gly, Pro aromatic Trp, Tyr, Phe The variant polypeptides can be
made using
methods known in the art such as oligonucleotide-mediated (site-directed)
mutagenesis,
alanine scanning, and PCR mutagenesis. Site-directed mutagenesis (Carter,
1986; Zoller




and Smith, 1987), cassette mutagenesis, restriction selection mutagenesis
(Wells et al. ,
1985) or other known techniques can be performed on the cloned DNA to produce
the
SHAAGtide variant DNA (Ausubel et al. , 1987; Sambrook, 1989).

STEM CELL PROLIFERATION (as an aside we could put high levels of zinc into the
stem
cell nutrient solution/tissue culture/medium to increase appetite and faster
growth as well
Vitamin E is known for keeping seniors healthier and may help to keep stem
cells or microbe
cells to further proliferate...).

I) We plan to batch cells at different stages of stem cell development by
using protein kinase
enzymes to catalyze phosphate group from ATP to a target protein to activate
or deactivate
specific proteins.

We plan to pulse regulatory proteins called cyclins to dependent kinases to
work in rhythm
with MPF (maturation promoting factor) that will recycle the sequence of
events during cell
cycle.

As well stem cells that are full enough of cytoplasm to reach the threshold
volume to genome
ration, is also being tested.

We plan to use Bio Flip Chip (BFC), a microfabricated polymer chip containing
thousands of
microwells (for example if we centrifuge the microbes found in a termite's gut
and put into
well foring colonies and tissue culture then when there are multiple identical
cells (stem cells)
we simply screen the DNA of from a single cell from the colony the seed cell
would possess
(selected after verified testing) the phenotypes that are the most
productive/efficient cost and
difficulty and hardiness to produce butanol and survive the toxins), and with
gelatin, a 3-D
substrate, and even another layer of cells.

We are testing the effects of SHP2-mediated activation of the MAP kinase
cascade in
regards to proliferation of stem cells.

II) We could take a somatic DNA from a female (and/or female) and micro
inject, electro fuse,
and/or laser fuse/heat shock into any female's egg that has had its DNA
removed or a
fertilized egg that has had its DNA removed. The idea is that the cytoplasm
has reached its
threshold volume to genome ration, (eg. one theory is that there is enough key
organelles -
that can't be easily duplicated that exists in duplicate in the parent cell;
to migrate to each
polar end of the division).

III) We could protoplast fuse adult somatic cells (from where there are large
collections like
the skin and the gut) with stem cells that have stopped proliferating or
replace micro inject
adult DNA into stem that have stopped proliferating and have had their DNA
removed, and
electrofuse/laserfuse/heat shock, the cytoplasm to fuse to the new DNA,
perhaps the adult
DNA is longer, thinner than the stem cells' own DNA which have been continuing
to divide for
some time (and therefore having inadequate inputs (in the necessary conditions
eg. DNA
length and width) to provide for the process to naturally and properly
complete into further
stem cell proliferation via chromatid duplication... We might also try micro
injection removal
of cytoplasm from an egg (fertilized and/or fertilized), suck up the top pole
and lower poles'
cytoplasm separately and micro injection remove either and adult cell
(somatic) and/or stem
cells that have stopped proliferating - most of its cytoplasm, leaving the DNA
intact then
reinjecting the cytoplasm from the egg into the top and the lower pole of the
cells that are no
longer proliferating until the cytoplasm has reached its threshold volume to
genome ration.



Another option is to take stem cells that have stopped proliferating and
inject trying both top
and bottom cytoplasm injection (where the cytoplasm comes form any source -
though we
think the most reproductive cytoplasm come form the egg), to replenish the
cytoplasm
material in the proper distribution, so that the two new cells both contain
the main organelles
structures distributed evenly between the two, while micro injecting cytoplasm
allows the
cytoplasm to reach it volume to genome ratio.

We could microinject/electro fuse/laser fuse/heat shock human DNA into pigs
and fast
dividing bacteria and/or lamas emus. Or better yet

First we tackle the ability of microbes (yeast) (especially the strong
performing - measured
by the durability of the cellulose) to breakdown (eg. cow's gut, that breaks
down straw);
found in termites' and/or cow's and/or lamb's and/or goats' and any and all
digestive systems
that are good at breaking down cellulose guts.

Second, we mix these corn (biocrops) and possibly supplements of vitamins and
necessary
nutrients not found in the bio crops that are important to the health and
productivity as well as
oxycyte or other artificial blood to deliver oxygen, nutrients and remove
carbon dioxide and
regulating PH and also regulating temperature - possibly by cycles of the
microbes to
produce butanol. We take the microbes that survive the butanol toxins (given
this proven
track record - the secret is to isolate and breed microbes that 1. survive
butanol toxins and 2.
are the most productive in turning bio crops into biofuel; both in time (how
fast), and ration of
inputs (costliness) to outputs. We can tissue culture the optimal microbes
and/or inject their
DNA into other microbes that are plentiful and prolific or non proven but
easily available
(removing the host cells' DNA in the case that these optimal microbes stop
proliferating and
then adding the proven DNA into unproven or more prolific or more plentiful
host cells that
have had their DNA removed). And/or we could protoplast fuse the cells with
the best
phenotypes (prolific, hardy - not requiring constant/difficult care including
survival in butanol
toxins, productive, efficient. Furthermore using the stem cells (tissue
culture) stock of termite
and/or cow gut microbes (are proven) to survive the butanol toxins, and are
effective
(productive) and hardy, we could identify the types of microbes and study
their genes in
comparison to the less successful genes to identify the type the genes that
make the good
microbes versus the unwanted microbes. As well the microbes that are more
effective and
productive, could be used to produce their enzymes, whether organically
stimulated and/or
harvested for their molecular make-up and artificial synthesis.

One option is to gather a good number of microbes from the termites' and/or
cows' gut, then
centrifuge the microbes, tissue culture and/or each cell into colonies (and/or
stem cells if
necessary; if can serve an advantageous purpose) then use a assembly line (eg.
electron)
microscopic viewer (perhaps with thermal sensor - if we need to see inside the
makeup of
the cells, the content, organelles, shape, sizes to identify them) and
Artificial Intelligence or
computer that can pick out and identify differentiating microbe cells by
running the tests
below... 1) common reactions, 2) antibody tests, 3) DNA, PCR probes and
primers.

A Brookhaven team has developed just such a technique, which they call "single
point
genome signature tagging." Using enzymes that recognize specific sequences in
the genetic
code, they chop the microbial genomes into small segments that contain
identifier genes
common to all microbial species, plus enough unique genetic information to
tell the microbes
apart.



In one example, the scientists cut and splice pieces of DNA to produce "tags"
that contain 16
"letters" of genetic code somewhat "upstream" from the beginning of the gene
that codes for
a piece of the ribosome - the highly conserved "single point" reference gene.
By sequencing
these tags and comparing the sequenced code with databases of known bacterial
genomes,
the Brookhaven team determined that this specific 16-letter region contains
enough unique
genetic information to successfully identify all community members down to the
genus level,
and most to the species level as well.

"Sequencing is expensive, so the shorter the section you can sequence and
still get useful
information, the better," van der Lelie said. "In fact, because these tags are
so short, we
'glue' 10 to 30 of them together to sequence all at one time, making this a
highly efficient,
cost-effective technique."

For tag sequences that can't be matched to an already sequenced bacterial
genome (of
which there are only a couple hundred), the scientists can use the tag as a
primer to
sequence the entire attached ribosomal gene. This gene is about 1400 genetic-
code-letters
long, so this is a more time-consuming and expensive task. But since ribosomal
genes have
been sequenced and cataloged from more than 100,000 bacterial species, this
"ribotyping"
technique makes use of a vast database for comparison.

Then test out the (strengths) effectiveness of each type of microbe and the
specific enzymes
that each microbe produces, and to artificially synthesize (duplicate) the
enzyme in the
laboratory - eg. by its molecular makeup and the catalyst conditions of the
amino acids and
their areas of linkages - maybe eventually synthesizing enzumes that are even
more
productive and hardier).

Another possibility is to take ant microbes from the termites' stomachs and
try injecting their
DNA into either cow's microbes found in their stomach and/or grow the ants
microbes directly
in the cow's stomach, (where removal of other microbes in the cow's stomach
might be
warranted to reduce competition for proliferation) and/or create a synthetic
cow's stomach
and/or grow under tissue culture (eg. fibroblasts... ). We could create
artificial elements of
the termite or and/or cows' stomachs, including pressure, PH, temperature...to
breed the
microbes

Known micro-organisms such as bacteria, protozoa, and fungi enzyme feed
supplements
containing fungal cellulase produced by Trichoderma viride cellulase enzyme.

Using the same single point genome signature tagging technique, we could apply
it to stem
cells, we compare the genome of stem cells using enzymes that recognizes
specific
sequences in genetic code, then chop the stem cells' genes into small segments
that contain
identifier genes common to all close species (or of single individual to
single type of stem cell
to single stem cell) of the stem cells in question and compare stem cells'
genes at their most
proliferative and when they are no longer proliferative (but not yet
specialized), and we chop
off the genes that are common (eg. both activated and/or both deactivated) to
both the two
types of stem cells and focus on the genes that are active in one and inactive
in the other.
We could also these techniques for any and all health solutions by 1) ruling
out all segments
of (genetic related) diseases (bacteria and/or viruses) and/or desired
phenotype genetic traits
(eg. tall height, intelligence...), by by using enzymes that recognize
specific sequences of



genetic code, then chop the genes into small segments that contain identifier
genes common
to all close species (or a single individual to another individual or even the
same individual
where one cell has cancer and the other does not) we chop off the genes that
are common to
both then we focus on the genes that are active in one and the same gene is
inactive in the
other

Using enzymes that recognize specific sequences in the genetic code, they chop
the
microbial genomes into small segments that contain identifier genes common to
all microbial
species, plus enough unique genetic information to tell the microbes apart.

We could micro inject human DNA into pigs fertilized and/or unfertilized eggs
(of which the
host has had its DNA removed).

Tissue Culture Medium should include:
20 amino acids include

Essential. Aarginine, Histidine, Isoleucine, Leucine, Lysine, Methionine,
Phenylalanine,
Threonine, Tryptophan, Valine.

Non-Essential Alanine, Aspartic Acid, Citruiline, Cystine, Glutamic Acid,
Glycine, Proline,
Hydroxyproline, Serine, Tyrosine

Vitamins A (vision, bone growth, epithelial growth), D (bone growth), E
(membrane structure,
antioxidant, immune functions), K (blood clotting), B1 (carbohydrate and fat
metabolism), B2
(energy metabolism and normal growth), Niacin (co-enzyme for energy
metabolism)

Major needed minerals are - Ca, P, Na, K, Mg, S, Cl (electrolytes and
metabolism)
Minor minerals - B, Co, Cr, F, Fe, Mn, SE, Si. . (co-enzymes and building
blocks of
proteins)

We are also protoplast fusing corn to jatropha, to produce the phenotype of
drought (low
water consumption) resistant crop whose mass (eg corn may be more productive,
eg input
to levels of outputs) and also the husk and cellulose because of the jatropha
may be more
easily fermented We will also try protoplast and/or/also micro injection DNA
into DNA ovum
cells that we tissue culture removed cells between the two plants above algae,
coconuts,
sugar cane, and palm oil We are looking at cloning corn and bio fuel plants
involving callus,
auxin, and cytokinin

We are looking at cloning implanting coffee DNA (from the animals that eat
selected coffee
bean - connoisseurs) into vanilla or other aromatic and flavourful seeds (that
are safe for
human consumption) Included are Colchicum seed, Thistle seed, Cumin seed, Anis
seed,
Coriander seed, Poppy seed, Pepper seed, Thyme seed,

Species
Coffea arabica -
Arabica Coffee
Coffea



benghalensis -
Bengal coffee
Coffea canephora
- Robusta coffee
Coffea congensis
- Congo coffee
Coffea dewevrei -
Excelsa coffee
Coffea excelsa -
Liberian coffee
Coffea gallienii
Coffea bonnieri
Coffea mogeneti
Coffea liberica -
Liberian coffee
Coffea
stenophylla -
Sierra Leonian
coffee

Types of beans
.cndot. Vicia
~ Faba or broad bean


Image

Vica faba or broad beans, known in the US as fava beans
.cndot. Vigna
~ Aconitifolia or Moth bean
~ Angularis or azuki bean
~ mungo or urad bean



~ radiata or mung bean
~ umbellatta or rice bean
~ unguiculata or cowpea (includes the black-eyed pea, yardlong bean and
others)
.cndot. Cicer
~ arietinum or chickpea
.cndot. Pisum
~ sativum or pea
.cndot. Lathyrus
.cndot. Lathyrus sativus (Indian pea)
.cndot. Lathyrus tuberosus (Tuberous pea)
.cndot. Lens
~ culinaris or lentil


Image

Lentils

.cndot. Lablab
~ purpureus or hyacinth bean
~ Phaseolus
~ acutifolius or tepary bean
~ coccineus or runner bean
~ lunatus or lima bean
~ vulgaris or common bean (includes the pinto bean, kidney bean and many
others)
.cndot. Glycine
~ max or soybean
.cndot. Psophocarpus
~ tetragonolobus or winged bean

Image

Psophocarpus tetragonolobus (winged bean)



.cndot. Cajanus
~ cajan or pigeon pea
.cndot. Stizolobium
~ spp or velvet bean
.cndot. Cyamopsis
~ tetragonoloba or guar
.cndot. Canavalia
~ ensiformis or jack bean
~ gladiata or sword bean
.cndot. Macrotyloma
~ M. uniflorum or horse gram
.cndot. Lupinus or Lupin
~ L. mutabilis or tarwi
.cndot. Ervthrina
~ E. herbacea or Coral bean

We could use different sound vibrations to stimulate responses for any and all
living
organisms. The sound can be pulsed in micro and macro rthyhm/cyclical.

For example in the case of stem cell proliferation, if we could 1) record the
vibrations made
by the cells at their most proliferative - and healthy (eg. blastocyst) stage
the replay when
their proliferation has stopped, 2) trial and/or error of various sounds (eg.
woofer), genre of
music, chanting.

Also the sound could be recorded from the human body (or neural; mind sound;
electrode
receptors) when they overcome disease, and then replayed for patients who
might benefit
from these bodily vibrations and/or trial and error of various sound, genre of
music, chanting;
these includes all disease such as cancer or even plaque on the teeth (less
effective at
corroding teeth and gums), if we could aim the sound at the area of infection,
and disrupt the
regulation of its processes (eg. proliferation and/or metabolism...), inhibit
spreading and
inhibiting serious harm (additionally to treat cancer we could inject enzymes
into a retro virus
that interacts with the cancer cells and wreaks havoc with the cancer cells
within its cell
membrane)

We could record a colonies' sounds for desirable microbes (could some microbes
be
complementary in nature, requiring the active - eg. cell signalling, products
of one microbe to
make another microbe more effective) when they were healthy - without the
stress of the
butonal toxin.

Additionally, we could record the sound made either a fetus (growing limbs
from the
blastema and/or stem cells) and/or a salamanders' sound when regenerating its
limb and/or
trial and error of sounds to help amputees to grow new limbs (see below).

Perhaps humans could regrow limbs and organs by growing using a blastema (stem
cells)
with the Human HOX genes and pressure that is found in that area of the body
as well as cell
signalling, with new doses of stem cells to be added as the stem cells migrate
and form the
different stages of embryonic development of the limb and/or organ.

The other option is to inject the stem cells together with activated HOX genes
regularly
(perhaps by Intra Venous drip GP; GV) and in time before scarring even use
topical and/or



abrasive dermal removal of scar tissue, even shave of the overgrown tissue in
preparation
for the mixture of stem cells with activated HOX gene to create a human
blastema as well as
cell signalling. The stem cells with activated HOX genes can be added directly
to the (eg.
removed area - limb) as the progress of migration and distal tip, stage by
stage
embryogenesis converts and/or causes the stem cells to produce the limb/organs
that is just
in time method, so there is no excess or not enough stem cells with the
activated HOX genes
to cause unwanted growth or in the case of not enough administration scar
tissue may form
or the regeneration may be instructed to stop due to lack of stem cells with
activated HOX
genes by whatever the body's recognition is, ...and also where the best place
to administer
the concoction, and amounts...

Furthermore through any and/or all gene therapies we could activate the HOX
genes in stem
cells to further proliferate them.

Humans generally contain homeobox genes in four clusters:

Image


There is also a "distal-less homeobox" family: DLX1, DLX2, DLX3, DLX4, DLX,
and DLX6.

"HESX homeobox 1" is also known as HESX1.

Short stature homeobox gene is also known as SHOX.


Tissue culture stem cells in direct contact with BMEC monolayers, or
physically separated by
microporous membrane. In unstimulated condition BMEC monolayers constitutively
produce
interleukin-6, Kit- ligand, granulocyte colony-stimulating factor, and
granulocyte macrophage
colony-stimulating factor.

Microbes from a pig's stomach and/or a vulture's stomach (that can survive,
thrive even in
such a rotten toxic environment) for their enzymes to process organic,
restaurant waste,
rotten organic garbage, offal, (especially manures and sewage) and compost as
well. Some
areas to start with include; Actinomyces bovis, Lactobacillus cellobiosus,
Micrococcus luteus,
Neisseria sicca, Clostridium bifermentans, Enterobacter agglomerans,
Peptostreptococcus




sp., Sarcina sp., Serratia odorifera, and Shigella flexneri Lactobacillus
acidophilus,
Streptococcus faecium, and Saccharomyces cerevisiae. We could also test these
microbes
with cellulose and producing butonal or any and all biofuels. We could also
test the
effectiveness of termite and cow gut microbes in processing organic wastes
(eg. eg. sewage
and manures). We could combine the rotten toxic waste microbes of vultures and
swine with
the cellulose enzyme break down of cows and termite microbes and apply to both
manure
and cellulose digestion. We could protoplast fuse such as Bio Flip Chip (BFC),
a
microfabricated polymer chip containing thousands of microwells that align
together two
types of microbes (selected based on ideal phenotype criteria, such as
vulture's gut microbes
hardiness to toxins and termite's voracious ability to break down even wood).

We are testing the use of .beta.-glucanase and pentosanases mixed to degrade
the .beta.-glucans
and pentosans (complex carbohydrates that interfere with digestibility of
other nutrients), in
grains (especially corn)...

We are testing phytase for it's enzyme that cleaves the ortho-phosphate groups
from phytic
acid (phytate), the major source of phosphorus in cereal grains and oilseed
meals (and/or all
bio fuel crops, eg. corn, and/or organic wastes..including wood waste).

probiotics,"
Some enzymes we will test specifically in the molecular design of cellulases,
hemicellulases,
and proteases of a form (related and/or mutated) that come from microbes that
are the best
performing (see the criteria above).

We will also lower the PH. . I believe to be conducive for good microbes to
flourish.

Citric acid, fumaric acid, or formic acid are used in digestion; probably by
helping the good
bacteria thrive.

Inorganic acids, such as phosphoric acid and, in some instances, hydrochloric
acid, also
have been found to help digestion.

We are experimenting SCFAs with saccharolytic fermentation which involves
acetic acid,
propionic acid, and butyric acid. Gases as well as organic acids like lactic
acid are also
produced by saccahrolytic fermentation. Acetic acid is used by muscle,
propionic acid helps
the liver produce ATP, and butyric acid provides energy to gut cells and may
prevent cancer
(University of Glasgow. 2005, Gibson RG. 2004, Beaugerie L and Petit JC.
2004).

We are already tissuing culturing cells (microbes) in:

We then set up the medium we are considering several alternatives or
combination (even all
together).

(Passaging of cells are used to further proliferate stem cells) Accutase
(Innovative Cell
Technologies Inc., distributed by PAA) and triturated approximately 10 times
using a pipette.
Then, the cell suspension was incubated. Dissociated cells were centrifuged at
120 rcf and
resuspended. An aliquot was counted by trypan blue exclusion assay in a
hemocytometer to
determine the amount of viable cells. Cells (105) were plated in T75 culture
flasks for long-



term passaging (culture medium (coated with fibronectin/lanmine) per flask).
All media
contained 1-glutamine, penicillin/streptomycin, heparin (Sigma), bFGF (R&D
Systems), and
EGF (R&D Systems). A total of 104 cells per well were seeded in 12-well plates
in a volume
of 1 ml and grown under standard conditions. After a few days the grown stem
cells were
counted and dissociated by Accutase, and viable cells were counted by trypan
blue exclusion
assay in a hemocytometer.

One of the problem we foresee, after identyfing then producing the microbes
enzymes easily,
in large amounts and cheaply is that corn husks and leaves is fiberous so it
is harder to
digest, stretching the time needed to digest, these microbes may go hungry and
start to
starve if they aren't getting enough nutrients over the time it takes to
digest the fiberous
cellulose - if this is the case we may have to mix in carbohydrates and other
nutrients found
in the cell culture medium above.


Image

Treatment Options - Treatment Information

During treatment for intestinal yeast and/or bacterial overgrowth it is
essential that probiotic
bacteria are consumed concurrently to restore the proper balance of organisms
in the gut.
When you undergo anti-fungal and/or anti-bacterial therapy when the pathogenic
organisms
are killed off space within the intestines and along the intestinal wall
becomes available for
colonization by other organisms. Taking probiotic supplements enhances the
chances of
these new colonies being made up of beneficial bacteria rather than more
pathogenic types
Also of importance to sufferers of environmental illnesses is the fact that
recent research
has shown that the gut flora is directly linked to the development of
allergies to both food
and airborne allergens and that improving gut flora could potentially reduce
the number and
severity of allergies1,2.

At first the range of probiotic products on the market will likely seem
overwhelming and you
won't know whether you are getting a good product or not until you learn a
little bit more
about the subject. The information below will explain a bit about probiotic
bacteria,
otherwise known as'beneficial bacteria' or 'friendly bacteria', and provide
some basic
pointers on what to look for in a probiotic product.

Types of Probiotic Bacteria

The most numerous arobiotic bacteria normally inhabiting the small intestine
are species of



Lactobacilli. In the colon the majority are mainly Bifidobacteria. Most
probiotic products
consist of one or more species of bacteria from one or both of these types.
Some products
available mainly in europe may also contain certain beneficial species of
E.coli but these
are rare at this time.


Let's take a look at some of the most well researched probiotic bacteria and
those found
most commonly in probiotic supplements.


Lactobacillus Acidophilus
By far the most well known species of probiotic bacteria is Lactobacillus
acidophilus which
has led many people to refer to probiotics simply as "acidophilus". This
status is not without
reason as the acidophilus species is the most prevalent in the human intestine
and has
been the most widely studied probiotic bacteria with research starting on
L.acidophilus as
long ago as 1925. The best researched single strain of acidophilus is probably
the DDS-1
strain. This strain amongst other benefits has been shown to stimulate the
immune system,
(WE COULD USE THIS BACTERIA FOR HIV/AIDS PATIENTS AND OTHER IMMUNE
DEFFIENCY PROBLEMS ANYWHERE WHERE IMMUNE SYSTEM IS COMPROMISED)
increasing levels of interleukin-1 alpha (IL-1 alpha) and (WE COULD USE THIS
BACTERIA
KEEP STEM CEALLS FROM DIFFERENTIATING - WHERE ONE POSSIBLE THEORY IS
THAT IF THE STEM CELLS DON'T DIFFERENTIATE THEY WILL CONTINUE TO
PROLIFERATE AND ALSO TO BALANCE OUT THE DANGERS OF STEM CELLS
BECOMING TUMOURS) (WE COULD ALSO USE THIS BACTERIA MULTIPLE NEEDLE
MICRO INJECT AND/OR RETRO VIRUS IT INTO THE TUMOURS) (ANOTHER USE IS
TO TREAT THE DIFFERENTIATED STEM CELL BACTERIA BEFORE THE CELLS
ENTER THE BODY TO COUNTER ANY CANCER TENDENCIES) (FINALLY WE COULD
EITHER MULTIPLE MICRO INJECT AND/OR DELIVER VIA RETRO VIRUS FILLED WITH
THIS BACTERIA AT THE SAME TIME A CHEMOTHERAPY PERHAPS EVEN IN THE
SAME RETROVIRUS SHELL TO THE CANCER AREA) tumour necrosis factor-alpha
(TNF-alpha) which suppress cancerous tumour growth3. Other research has shown
that
L.acidophilus DDS-1 also alleviates lactose intolerance by producing
significant amounts of
the lactose digesting enzyme lactase, inhibits gastrointestinal pathogens by
producing
antimicrobial substances such as acidophilin and helps alleviate dermatitis
and other skin
conditions by altering gut flora amongst other things.


Lactobacillus Rhamnosus
Lactobacillus rhamnosus is a probiotic bacteria that has been receiving a
growing amount of
attention as a treatment for many illnesses in the form of the GG strain.
Lactobacillus
rhamnosus GG (LGG) now has a wealth of research backing its use, particularly
for
infectious and allergic conditions. A 2001 study reviewing information
regarding probiotics
and infectious diseases found that there is a large amount of data showing
that
Lactobacillus GG is an effective treatment for diarrhea caused by Clostridium
difficile
infection4. Another study testing the effectiveness of probiotics in
preventing allergic illness
found that Lactobacillus GG given prenatally to mothers with at least 1 first-
degree relative
and postnatally for 6 months to their infants reduced the incidence of atopic
eczema by half




compared to controls5.


Lactobacillus Bulgaricus (Galactooligosaccharides PREBIOTICS FACILITATES THIS
PROBIOTIC)
This organism is slightly different to most probiotic bacteria in that it is a
'transient bacteria'.
It is referred to in this way because unlike most probiotic bacteria it
doesn't adhere to the
intestinal wall and form colonies, rather it simply passes through the
digestive system and
leaves the body in the stool. It has many beneficial effects as it passes
through the digestive
tract, however. These include enhancing the digestibility of milk products and
other proteins
and producing natural antibiotic substances that specifically target
pathogenic bacteria
whilst sparing friendly species. In this sense L.bulgaricus can be thought of
as a helper to
colonizing bacteria just as the immune system has T helper cells to support
other immune
cells. A study published in the World Journal of Gastroenterology showed that
L.bulgaricus
could suppress inflammatory immune reactions in the intestinal wall thus
preventing tissue
damage6. In another study a substance produced by (CAN BE USED DEFENSE AGAINST

PATHOGENIC ORGANISMS AND MIXED INTO THE MICROBES USED TO BREAK
DOWN BIOFUEL ORGANIC WASTE (EG. CORN HUSKS) AND SEWAGE AND OTHER
SUCH RELATEED ORGANIC WASTE - HOPEFULLY NOT ATTACKING THE MICROBES
WITH THE MOST WANTED PHENOTYPES, DOING MORE GOOD THAN HARM)
L.bulgaricus was shown to stimulate activity in part of the gut immune system
called the
Peyer's patches which provide defense against pathogenic organisms within the
gut7.

Lactobacillus Salivarius
L.salivarius has been repeatedly shown to inhibit the bacteria Helicobacter
pylori (H.pylori)
which is responsible for the creation of peptic ulcers8. H.pylori interferes
with stomach acid
production and/or produces a toxin that directly contributes to ulcer
formation. The usual
treatment involves taking antibiotics but H.pylori may become resistant to
them and there
are side effects of prolonged antibiotic use due to the eradication of
beneficial bacteria
along with the pathogenic bacteria. It has been discovered that L.salivarius
produces large
amounts of lactic acid that completely inhibits the growth of H.pylori and
reduces the
associated inflammatory response. The first bacteriocin (natural antibiotic
substance) to be
isolated and studied at the genetic level was taken from a strain of
L.salivarius9.

Lactobacillus Plantarum
This bacteria is the most prevalent species in most naturally fermented foods.
It has the
ability to block receptor sites for gram negative bacteria and so is effective
as an antibiotic.
It is an important player in (WE COULD USE THIS BACTERIA FOR ANY AND ALL




IMMUNE DEFIENCY PROBLEMS ANYWHERE WHERE THE IMMUNE SYSTEM IS
COMPROMISED) antimicrobial defense and is effective against both extra and
intracellular
pathogens. L.plantarum is also capable of digesting (WE COULD USE THIS PRODUCT
TO
BREAK DOWN BIO FUEL CROPS WASTE AS WELL AS WOOD WASTES) semi-
digestible fibres such as those found in onions, garlic, wheat, oats, rye and
yeast. It may
therefore help with digestive problems like gas and bloating. Recent research
has shown
that L.plantarum has the ability to break down bile acids and lower
cholesterol10 and is
extremely resistant to stress conditions including high temperature and
(ESPECIALLY
GOOD FOR BUTANOL PRODUCTION) concentrations of ethanol, extremes of pH and the

freeze drying process that would normally kill lactic acid bacteria11.


Lactobacillus Casei
This species is commonly found in probiotic dairy foods such as 'live
yoghurt', hence the
name 'casei' which relates to the milk protein casein. It was reported in
Microbiology and
Immunology to have the most potent protective activity against the Listeria
bacteria. Listeria
is potentially lethal with about 30% of victims dying. It is most commonly
transmitted through
consumption of dairy products and raw vegetables. Like L.salivarius, L.casei,
in the form of
the shirota strain found in Yakult probiotic yoghurt drinks, has been shown to
significantly
inhibit the growth of the peptic ulcer causing bacteria H.pylori12. A
probiotic drink containing
the shirota strain has also been shown to reduce the severity of constipation
as evidenced
by both patient response to questionnaires and physical examinations13.
Finally, a study
with malnourished mice showed that L.casei (combined with FOS), when given
along with a
re-nutrition diet, enhanced the immune response and increased resistance to
certain
pathogenic bacteria in the digestive tract14.


Lactobacillus Sporogenes
In a study at the G.B. Pant hospital in New Delhi, India, Lactobacillus
sporogenes was able
to lower cholesterol levels by 104 points. It produced a highly significant
reduction in LDL
cholesterol ('bad cholesterol) levels and a small but significant increase in
HDL cholesterol
('good cholesterol'). This study offers the prospect of using L.sporogenes as
a side-effect
free alternative to drug therapy in the treatment of high cholesterol and
heart disease. In a
multi-centre double-blind placebo controlled trial, L.sporogenes was found to
be nearly
twice as effective as placebo in reducing the number of episodes and duration
of diarrhea
following antibiotic treatment in children15. As well as being used to lower
cholesterol,
Alternative Medical Review reports that L.sporogenes has been used in the
treatment of gut
dysbiosis, vaginitis and aphthous stomatitis16.


Bifidobacteria Bifidum (Galactooligosaccharides PREBIOTICS FACILITATES THIS
PROBIOTIC)
This bacteria is one of the major constituents of the normal flora in the
colon and is the most
common Bifidobacteria species found in probiotic products. It is reportedly
well tolerated,
reduces the (inflammatory response (SEE INFLAMMATORY DISEASES BELOW) in the
colon and (COULD BE USED WITH hiv/aids AND OTHER IMMUNE PROBLEMS WHERE
IMMUNE SYSTEM HAS BEEN COMPROMISED) stimulates the body's fluid immunity. A




study carried out at the Women and Children's Hospital of Buffalo, NY showed
that
B.bifidum can significantly reduce the intestinal concentration of endotoxin,
which is made
up of the cell walls of (WE COULD USE THIS BACTERIA ON STEM CELLS TO CLEAN
OFF THE THECELL WALLS SO THE STEM CELLS CONTINUE TO PROLIFERATE) (WE
COULD ALSO USE THIS PRODUCT TO CLEAN OFF SCARRING TISSUE) dead bacteria
and is toxic if allowed to build up17. In another study B.bifidum of human
origin was found to
adhere well to the intestinal wall and significantly reduce the ability of
pathogenic E.coli to
do the same (WE COULD ALSO BE USED ON PATIENTS WHOSE CANCER IS IN
REMISSION TO CLEAN OUT THE REMAINING TOXICITY FROM THE REMAINING
CHEMOTHERAPY)18. Research carried out by the Yakult company who manufacture
probiotic drinks showed that their patented strain of B.bifidum had
significant (WE COULD
USE THIS PRODUCT IN THE MEDIUM AND/OR MICRO INJECT INTO THE CELL
ESPECIALLY THE NUCLEUS TO TRY TO KEEP STEM CELLS PROLIFERATING) anti-
oxidant action and was able to protect the intestinal lining from lipid
peroxidation in iron
overloaded mice19.


Bifidobacteria Longum (Galactooligosaccharides PREBIOTICS FACILITATES THIS
PROBIOTIC)
B.longum is another species of Bifidobacteria commonly found in probiotic
products. It is
reportedly able to eliminate the (CAN BE USED TO CLEAN OUT NITRATES FROM THE
WATER TABLE NEAR FARMS) nitrates commonly found in foods ingested by humans.
Levels of nitrate commonly ingested by humans are unable to kill this species.
B.longum
has been shown to inhibit the action of vero cytotoxin produced by some
strains of E.coli
which can cause hemorrhagic colitis and hemolytic uremic syndrome in humans.
(WE ARE
LOOKING AT USING THIS BINDING TO TOXIN EFFECT FOR BINDING TO ANY AND
ALL TOXINS, POISONS...USEFUL IN SEWAGE TREATMENT). It achieves this by
producing substances that bind to the vero cytotoxins20. B.longum has also
been shown to
have a protective effect against infection with Salmonella Typhimurium,
possibly due to an
anti-inflammatory action21 (SEE INFLAMMATORY DISEASES BELOW).

Bifidobacteria Infantis (Galactooligosaccharides PREBIOTICS FACILITATES THIS
PROBIOTIC)
B.infantis is known to have an inhibitory action on invasive pathogenic
bacteria such as
E.coli. Research has shown that it achieves this inhibition through more than
one
mechanism which explains why it is effective against a range of pathogens.
Inflammatory
bowel disease (IBD) is thought to be caused by organisms called bacteroides
which are a
normal component of the gut flora. B.infantis has the ability to highly reduce
the growth
of bacteroides and also significantly inhibit the inflammatory response (see
inflammatory diseases below) caused by them in the gut lining22. Other
research using
formulations containing B.infantis has found it to useful for treating
irritable bowel syndrome
(IBS) and diarrhea. Of all the strains of bacteria in the formulation
B.infantis was found be
one of the species that had colonized the intestines of patients to the
highest degree23.

Streptococcus Thermophilus




Like L.casei, this bacteria has been shown to (COULD STEMS STOP IN THEIR
PROLIFERATION BECAUSE THEY HAVE BEGUN TO ATROPHY - IF YES THEN
PERHAPS THIS BACTERIA MAY WORK TO AID FURTHER STEM CELL
PROLIFERATION) aid recovery from malnutrition due to short-term fasting and
reduce the
associated intestinal atrophy in animal studies24. S.thermophilus is also
known to have
powerful antioxidant activity, protecting the body from dangerous free
radicals which
increase in the body due to aging, stress, sugar, antibiotics and other
chemicals and toxins
It has also been shown to have (WE ALSO THEORIZE THAT PERHAPS THIS
BACTERIA'S ABILITY TO FIGHT CANCER OF WHICH WE ARE CLAIMING ITS USE FOR
ALL CANCERS, ADDITIONALLY WE WILL TRY TO USE THIS BACTERIA TO CAUSE
CELLS FROM DIFFERENTIATING INTO CANCER CELLS USING THIS MECHANISM TO
PREVENT STEM CELLS FROM DIFFERENTIATING LEAVING PLURIPOTENT AND
FURTHERMORE ABLE TO CONTINUE TO PROLIFERATE) (ANOTHER USE IS TO
TREAT THE DIFFERENTIATED STEM CELL PRODUCT BEFORE THE CELLS ENTER
THE BODY TO COUNTER ANY CANCER TENDENCIES) (FINALLY WE COULD EITHER
MULTIPLE MICRO INJECT AND/OR DELIVER VIA RETRO VIRUS FILLED WITH THIS
BACTERIA AT THE SAME TIME A CHEMOTHERAPY PERHAPS EVEN IN THE SAME
RETROVIRUS SHELL TO THE CANCER AREA) anti-tumour activity which is especially
effective against colon cancer cells.


Homeostatic Soil Organisms (HSO's)
In recent y ears a different sort of probiotics have become available known as
homeostatic
soil organisms. These are organisms that live naturally in the soil and used
to be ingested
regularly by humans before intensive farming methods removed them from the
food supply.
Most of the organisms found in HSO supplements are transient bacteria meaning
they don't
colonize the intestines but pass through, providing a number of benefits to us
as they do so.
Some of these benefits include aggressively killing pathogens, producing
specific antigens
that act to stimulate the immune system, create superoxide dismutase (SOD) a
powerful
antioxidant enzyme and help the body to metabolize proteins and eliminate
toxins. Some of
the benefits over traditional probiotic supplements that HSO's are said to
have are there
superior ability to survive stomach acid, ability to survive in any intestinal
pH and their
powerful ability to fight off infections in the GI tract. Many doctors and
patients have
reportedly had very good results using HSO's but as yet there is little good
scientific
research regarding their use.


Choosing a Probiotic Product


Unfortunately choosing a probiotic supplement is not as simple as just picking
up the first
bottle you find and assuming it will do the job. Some products contain
bacteria that are not
even known to be normal inhabitants of the human gastrointestinal tract and
the bacteria in
many have not undergone any testing with regards to their ability to colonize
once they
reach the intestines. There are a number of factors that will determine the
effectiveness of a




certain bacteria as a probiotic. Some of these are:

.cndot. Ability to survive the manufacturing process.
.cndot. Ability to survive heat, light, moisture etc during the time from
packaging to use.
.cndot. Ability to survive stomach acid
.cndot. Ability to attach to the intestinal wall
.cndot. Ability to fend off other organisms, survive in the current intestinal
environment and
successfully colonize.


As you can see, the fragile probiotic bacteria have a lot to deal with if they
are to eventually
colonize your intestines. It would be naive to assume that every species and
strain of
Lactobacilli and Bifidobacteria would be equally effective as probiotics. As a
result it is best
to research a product before you buy. Look out for the species of bacteria
covered on this
page but if you see a species that isn't covered listed on a supplement bottle
then do some
research on that bacteria online yourself to make sure it will be effective.
As a rule it is best
to look for products that state the specific strains of the bacteria they
contain such as
L.acidophilus DDS-1 and L.rhamnosus GG, where the DDS-1 and GG respectively
are the
strains. You can then look for research carried out with that particular
strain to assess its
effectiveness. The best place online to look for scientific research on
probiotic bacteria, or
any subject, is the PubMed database which can be found here.


Also of importance is the ability of a products packaging to protect the
bacteria from
environmental factors such as light, heat, moisture and oxygen. Dark coloured
glass bottles
protect well from light and heat. Some products use rubber caps under the
screw top of the
bottle, this offers added protection from oxygen and moisture by providing an
airtight seal.
However the best protection is offered by products where each dose is
individually
packaged in a foil sachet. This avoids the situation with a bottle where all
the
capsules/tablets are exposed to the environment every time you take the top
off. Also of
importance is how the product is stored in the store. If a product says it
needs to be
refrigerated, make sure this is the case in the store.


Following these guidelines should increase your chances of purchasing an
effective
probiotic supplement.


Prebiotics




Where probiotics are the beneficial bacteria found in the intestines
'prebiotics' are special
indigestible carbohydrates known as oligosaccharides that feed probiotic
bacteria and
encourage their growth. Oligosaccharides are found naturally in certain fruit
and vegetables,
including bananas, asparagus, garlic, wheat, tomatoes, Jerusalem artichoke,
onions and
chicory. Because of the ability of prebiotics to encourage the growth of
beneficial bacteria it
is worth considering a supplement when there is a need to improve the gut
flora. Prebiotics
can be taken on their own or with a probiotic supplement. They have an
advantage over
probiotic supplements in that there is no concern about oligosaccharides being
destroyed
while in storage or en route to the intestines through the stomach acid and
digestive
enzymes.


The most common types of prebiotics available in supplements are
fructooligosaccharides
(FOS), inulin and galactooligosaccharides. As well as, or perhaps due to,
encouraging the
growth of beneficial bacteria in the intestines, prebiotics have been shown to
have a
number of other benefits.

Fructooligosaccharides (FOS)
FOS has been shown in one study in mice to increase intestinal Riga, the
body's first line of
defense against invaders25. In another study using pigs, FOS was shown to
increase
butyrate concentrations in the large intestine26. Butyrate is a short chain
fatty acid (SCFA)
that helps to maintain the health of the colon.


Inulin
German research has shown that inulin is effective in improving the
composition of the gut
flora and reducing the severity of colitis symptoms when tested in rats27. In
a study
assessing the role of common foods in improving intestinal health cheese which
contains
inulin was found to have a beneficial effect, reducing bacteria and chance of
infection 28.

Galactooligosaccharides
In animal studies galactooligosaccharides have shown very promising results
increasing
populations of both lactobacilli and bifidobacteria and increasing beneficial
short chain fatty
acids29. A mixture of galacto and fructooligosaccharides added to standard
infant formulas
has has also been shown to increase both lactobacilli and bifidobacteria
species in human
infants30.


Probiotics and Prebiotics and be applied to:




Bifidobacteria Bifidum (Galactooligosaccharides PREBIOTICS FACILITATES THIS
PROBIOTIC)


Bifidobacteria Longum (Galactooligosaccharides PREBIOTICS FACILITATES THIS
PROBIOTIC)


Bifidobacteria Infantis (Galactooligosaccharides PREBIOTICS FACILITATES THIS
PROBIOTIC)


Diseases where anti inflammatory:

Multiple Sclerosis (MS)
Rheumatoid Arthritis (RA)
Inflammatory Bowel Disease (IBD)
Interstitial Cystitis (IC)
Fibromyalgia (FM)
Autonomic nervous dysfunction (AND neural-mediated hypotension);
Pyoderma Gangrenosum (PG)
Chronic Fatigue (CF) and Chronic Fatigue Syndrome (CFS).
Chronic hepatitis
Systemic lupus erythematosus
Arthritis
Thyroidosis
Scleroderma
Diabetes mellitus
Graves' disease
Beschet's disease and
Graft versus host disease (graft rejection).
Chronic inflammatory pathologies such as aneurysms
Hemorrhoids
Sarcoidosis
Chronic inflammatory bowel disease
Ulcerative colitis
Crohn's disease and vascular inflammatory pathologies
Disseminated intravascular coagulation
Atherosclerosis
Kawasaki's pathology
Coronary artery disease
Hypertension
Stroke
Asthma
Chronic hepatitis
Multiple sclerosis
Peripheral neuropathy
Chronic or recurrent sore throat
Laryngitis
Tracheobronchitis
Chronic vascular headaches (including migraines
Cluster headaches and tension headaches) and pneumonia





Neurodegenerative diseases including
Demyelinating diseasessuch as multiple sclerosis and acute transverse
myelitis;
Extrapyramidal and cerebellar disorders such as lesions of the corticospinal
system;
Disorders of the basal ganglia or cerebellar disorders;
Hyperkinetic movement disorders such as Huntington's Chorea and senile chorea;

Drug-induced movement disorders such as those induced by drugs which block CNS

dopamine receptors;
Hypokinetic movement disorders
such as Parkinson's disease;
Progressive supranucleo palsy;
Cerebellar and Spinocerebellar Disorders such as astructural lesions of the
cerebellum;
Spinocerebellar degenerations (spinal ataxia)
Friedreich's ataxia
Cerebellar cortical degenerations
Multiple systems degenerations (MencelDejerine-Thomas
Shi-Drager and Machado Joseph)); and systemic disorders (Refsum's disease
Abetalipoprotemia, ataxia telangiectasia and mitochondrial multi-system
disorder);
Demyelinating core disorders such as:
Multiple sclerosis
Acute transverse myelitis;
Disorders of the motor unit such as neurogenic muscular atrophies (anterior
horn cell
degeneration) such as
Amyotrophic lateral sclerosis
Infantile spinal muscular atrophy and juvenile spinal muscular atrophy);
Alzheimer's disease;
Down's Syndrome in middle age;
Diffuse Lewy body disease; Senile Dementia of Lewy body type;
Wernicke-Korsakoff syndrome;
Chronic alcoholism;
Creutzfeldt-Jakob disease;
Subacute sclerosing panencephalitis
Hallerrorden-Spatz disease; and
Dementia pugilistica

Malignant pathologies involving tumors or other malignancies such as:
Leukemias (acute chronic myelocytic
chronic lymphocytic and/or myelodyspastic syndrome);
Lymphomas (Hodgkin's and non-Hodgkin's lymphomas such as malignant lymphomas
(Burkitt's lymphoma or Mycosis fungoides));
Carcinomas (such as colon carcinoma) and metastases thereof;
Cancer-related angiogenesis;
Infantile hemangiomas;
Alcohol-induced hepatitis.
Ocular neovascularization
Psoriasis
Duodenal ulcers

Stem Cells Body Rejection:

Kupffer Cells, are the macrophages found in the sinusoidal lumen, attached to
the liver
sinusoidal endothelial cells, transplants of the liver have shown donor
Kupffer cells migrate to




the the recipient lymph nodes recipient derived monocytes also have been known
to replace
the opposite way. Kupffer cells play a role in regulating T lymphocte (not
only proliferation).
Furthermore in the case of liver transplants, I postulate that the stem cells
that are and can
be the precursor to Kupffer Cells, the Donor's DNA replaces the source stem
cells for the
Kupffer cells and thus regulation of the body's immune system, which produces
the majority
lymph in the body.

By destroying Kupffer cells in the Liver with intravenous application of
liposomeentrapped
dichloromethylene diphosphonate specifically hepatocytes and sinusoidal sites
on the liver
we observe the production of M-CSF and GM-CSF which regulate the
proliferation,
differentiation, and maturation of replacement Kupffer cells (and increase
neutrophils and
monocytes in response to the liposomeentrapped
dichloromethylene diphosphonate application) the theory is that hematoietic
stem cells
proliferate during the process in which the Kupffer cells are depleting. These
stem cells
being macrophage (lymph cell version in the liver known as - Kupffer Cells)
precursors were
derived from bone-marrow-derived and intrahepatic proliferating population.
Regarding the
increase in neutrophils and momocytes, as part of the outcome of the
liposomeentrapped
dichloromethylene diphosphonate application also, their proliferation in
synchronicity it is
believed that the residue of the destroyed macrophages with the application of

liposomeentrapped dichloromethylene diphosphonate one theory is that
leukocytosis is not
performed due to the lack of Kupffer cells during the depletion of Kupffer
cells stage of this
experiment.
It is believed that the substances resulting from the deaths of the Kupffer
cells might have
stimulated the bone marrow to release leukocytes and myeloid.
Focal Hematopoiesis is caused by glucan (application) to produce M-CSF and GM-
CSF
(together with IL-3), all of which help the number of Kupffer cells
reproliferate to the body's
replenishedment.
In Conclusion of this immune system phenomenon, the macrophage precursors
(Kupffer
cells) found after the application of liposomeentrapped dichloromethylene
diphosphonate
Were comprised not of intrahepatic repopulating cells, but more importantly
also bone-
marrow derived repopulating cells.
For the sake of immune system rejection inorgan transplant, could we seed
either the liver or
the bone marrow and/or both, with the organ donor's stem cells and
differentiated into
precursors of the immune cells (eg. lymphatic system cells...).
One way is to apply liposomeentrapped dichloromethylene diphosphonate to both
(or
separately to study the results) liver and the bone marrow (perhaps where the
bones have
the most precursor marrow type and apply at multiple sites possibly using
intravenous) and
then reseeding at multiple sites with stem cells (carrying the DNA cultured
from the donor of
the transplant organ) possibly mixed in layered for migration adherence to the
liver and bone
inner walls.
As substitutes for liposomeentrapped dichloromethylene diphosphonate, we plan
to use
chemotherapy (GP 45%) or total body irradiation (GP 1%), cyclophosphamide with
busulfan.
The novelty in our methods is that in addition to the organ (other than liver,
although liver and
bone marrow are probably always used together no matter what the transplant)
transplant,
we can grow a synthetic liver from outside the body, or grow graftable liver
tissue that are the
precursors, that proliferate into lymphatic cells when the immune system is
stimulated.
Theses precursor tissue could be derived from the organ (always including the
liver for liver
diseases but applicable for other organ transplant) donors' stem sells. At the
same time we
repopulate the bone marrow's precursor cells to that that it would normally
send to the liver to
replace/repopulate with stem cells as well as cells that are precursors to
immune system



cells derived from the donor organ's stem cells. Bone marrow stem cells
include
hematopoietic stem cells, which can be extracted from the donor's bones and
tissue cultured
and other proliferation techniques. The secret is that by destroying the
lymphatic cell
populations in both the liver and the bone marrow, and then replacing
(seeding) the pools of
areas where the patient's original stem cells once belonged with the donor's
of the transplant
organ stem cells and/or precursor to the immune system's cells the patient
will take on the
judgement call of the Donor's DNA, and not reject the new transplanted organ.

FARMING
USING the Tea Manure Processing for sewage in poor countries we could 1)
develop the
cattle/swine/chicken/bio fuel crops (any and all) farms underground, then suck
in the air from
the rising breaths and flattulence and Tea Manure processsing plants where the
manures are
collected and concentrate the methane (the methane is separated from carbon
dioxide via
DDR zeolite membrane strengthened by porous substrate such as titanium
(possibly ceramic
- especially if the methan is burnt nearby) and/or carbon nano
particles/tubes/fibers possibly
mixed with layers of clay, and/or combination of all mixed or in layers and/or
additionally we
could build TALL GRAVITY TO ELECTRICITY INVENTION surrounding the (Anyand all)

farms mixed with solar panels and wind farms above ground and/or underground.
We can
also use my mirrors to boiler (to boil urine and therefore concentratithe
nitrites in the urine) to
energy and fresh water (from salt water and/or urine). We can use probiotics
(mcirorganisms
- eg. bokashi) and other advantageous microorganisms (probiotics; prebiotics).
See below
for proper treatments for Heavy Metal Detox (taken from the Internet).

The most common source of heavy metal toxicity is from dental amalgam
fillings and other metal dental appliances. In 1989, the Environmental
Protection Agency (EPA) declared that amalgams are a hazardous
substance under the Superfund law. Scrap dental amalgam was declared a
hazardous waste in 1988 by the EPA. Outside of your mouth it has to be: 1.
Stored in unbreakable, tightly sealed containers away from heat. 2. It is not
to be touched. 3. Stored under liquid glycerine or photographic fixer
solution. So, once it is taken out of the mouth it is toxic, but when it is
placed in the teeth it is labeled "nontoxic." You can't throw it in the trash,

bury it in the ground or put it in a landfill, but they say it's okay to put
it in
people's mouths. It sounds like truth decay! Lead, mercury and cadmium
exert most of their toxicity by destroying important proteins, many of which
are enzymes, hormones, or cell receptors. Mercury will attach to sulfur
amino acid building blocks in proteins. The sulfur amino acids are
methionine, cysteine, and taurine. Sulfur is present in all proteins.
Numerous enzymes require intact sulfur groups and many are inactivated
by mercury.

Lead binds with the sulfur groups on proteins and inactivates them. Lead
suppresses neuron clusters in the brain, hindering brain development in
children by stunting the mapping of sensory nerves. One of the primary



ways the body gets rid of metal compounds is through a pathway that goes
from the liver into the bile where it is then transported to the small
intestine
and excreted in the feces. Inorganic mercury is complexed with glutathione
in the bile, suggesting that glutathione status is a major consideration in
the
biliary secretion of mercury. This same pathway is affected by a mercury
induced reduction of available taurine needed to produce bile acid
(taurocholic acid). When the microflora of the intestine has been reduced
through stress, poor diet, use of antibiotics and other drugs, fecal content
of
mercury is greatly reduced. Instead of being excreted in the feces, the
mercury gets recirculated back to the liver. The person that is under stress,
eating a poor diet, and/or taking antibiotics will tend to maintain a higher
body burden of mercury derived from dietary sources--especially if they are
eating diets high in fish.

Disposal of the body's burden of mercury is via the urine and feces,
although minute amounts are detectable in expired air. Excretion via the
liver occurs in bile and reabsorption of some of this mercury does take
place. However, the kidney is equipped with an efficient, energy-dependant
mechanism for disposing of metals such as mercury. Kidney tissue contains
a thiol-rich protein called metallothionein; exposure to toxic metals triggers

the production of this protein which binds tightly to the metal, retaining it
in
the kidney tissue in a relatively harmless form. As long as the kidney's
capacity for production of metallothionein is not overwhelmed, mercury
excretion can eventually balance intake, thereby limiting worsening of
symptoms. However, acute high doses of mercury, or an increase in the
chronic dose level can readily precipitate renal failure, one of the classic
symptoms of mercury poisoning.

Detoxification systems such as metallothionein, cytochrome P-450, and bile
are adversely affected by mercury. Metallothionein binds toxic metals in the
body to prepare them for excretion. Mercury ties up this material so it
cannot clear out other metals such as lead, cadmium, and aluminum.
Mercury from amalgam binds to -SH (sulfhydryl) groups, which are used in
almost every enzymatic process in the body. Mercury therefore has the
potential to disturb all metabolic processes.

A small proportion of total body mercury is excreted in various forms directly

in the urine without being bound to protein. In low dose, steady state
conditions, such as the dentist who has worked at a similar exposure level
for years, the urinary output very accurately reflects the total body burden
and this is why urine monitoring is so important.



The following is a list of nutrients that facilitate the removal of heavy
metals.
Mega H-: The negative hydride ions in Mega H- alter the water consumed
with the food and supplements in our diet, to have a lower surface tension
and an increased conductivity. A low surface tension in the extra cellular
fluids is also important in the removal of toxins from the cells and into
lymph
and venous blood for removal from the body. Tap water has a surface
tension of approximately 73 dynes/cm. The water around our cells has a
surface tension of approximately 45 dynes/cm. It is necessary, that the
body reduces the surface tension of water we consume in order for
nutrients to pass through cell walls, and for toxins to pass out of the cells.

Mega H- in water expedites this process. Glutathione: Contains cysteine,
glycine and glutamic acid. The liver manufactures glutathione whenever
extra cysteine is available. Blood glutathione levels change in direct
proportion to the amount of cysteine is in the diet. One 50 milligram capsule
or tablet, three times a day taken on an empty stomach. Individuals with
insulin deficiency should not take glutathione.

Methionine: Methionine levels are a major determinant in the liver's
concentration of sulphur-containing compounds, such as glutathione and
cysteine. As methionine is the precursor for the manufacture of cysteine in
the body, extra supplementation of this critical amino acid should increase
available cysteine. Animal studies have shown that methionine protects rats
from the toxic effects of lead and mercury. Chelating agents such as DMSA
(dimercapto succinic acid) and DMPS (dimercapto-propane sulfonic acid)
bind to cysteine for excretion. L-cysteine bound to mercury (L-penicillamine,
N-acetyl-L-cysteine, DMSA and glutathione complexed with methylmercury)
resembles the L-methionine molecule and can cross the blood brain barrier.
L-methionine inhibits the transport of these complexes into the brain.
Methionine increases the bioavailability of glutathione. Most of the cysteine
required for the resynthesis of glutathione must originate from methionine
and not from cysteine generated by the catabolism of glutathione. Patients
taking only D-L-methionine increased mercury excretion in the urine by 60%
over the excretion rate before taking the methionine. Lead excretion was
also increased. The L-form is rapidly metabolized by the liver and does not
offer a sustained antioxidant level. Over half of the D-form is slowly
metabolized by the same pathways as excess L, and acts identical to L as
an antioxidant. The benefit of the D-L form of methionine is the D form
provides sustained blood levels allowing he L-form to be converted to other
sulfur antioxidants. Babies need 22 mg/Kg body weight of methionine on a
daily basis while adults need 10 mg/Kg of body weight daily.



N-Acetyl-L-Cysteine (NAC): NAC forms L-cysteine, cystine, L-methionine,
glutathione (GSH), and mixed di-sulfides. Stimulates the body to produce
large amounts of cysteine and glutathione, thus greatly augmenting plasma
and red blood cell content of both cysteine and glutathione;
Methylsulfonylmethane (MSM): MSM, like fresh garlic, provides a
bioavailable dietary source of sulfur. MSM exerts a direct beneficial effect
in
ameliorating a variety of allergic responsees and pain associated with
systemic inflammatory disorders.

Milk Thistle (silymarin): Silymarin provides support and protection against
liver toxins which can cause free-radical-mediated oxidative damage.
Silymarin is many times more potent in antioxidant activity than vitamin E.
In addition, it increases liver production of glutathione and protects red
blood cell membranes against lipid peroxidation and hemolysis.
Chlorella: Is a food-like all purpose mild chelator of heavy metals; it is a
specially processed green-algae type of food that is taken with meals and is
quite tolerable and pleasant for many. But since chlorella is so easily
contaminated, the manufacturer's quality control is important. Nature's
Balance is a source of high quality chlorella that can be taken as a part of a

person'd detox program. The detoxification capability of Chlorella is due to
its unique cell wall and the material associated with it. The cell walls of
Chlorella have been shown to have three layers of which the thicker middle
layer contains cellulose microfibrils. Atkinson et al found a 14nm thick
trilaminar layer outside the cell wall proper which was extremely resistant to

breakage and thought to be composed of a polymerised carotene like
material.....Laboratory studies showed that there were two active absorbing
substances - sporopollenin (a naturally occurring carotene like polymer
which is resistant to degradation) and the algae cell walls." Chlorella's
ability to detoxify the body is very significant because of the large amount
of
chemicals we are exposed to in today's modern world. This ability to
detoxify chemicals is also one of the important differences between
Chlorella and other "green" products."

Cilantro: stimulates the body's release of mercury and other heavy metals
from the brain and CNS into other tissue. This facilitates the ability to
remove heavy metal from the body using other dietary protocols, such as
Chorella and other chlorophyll containing herbs such as Nettles and Alfalfa.
These herbs aid in detoxifying by denaturing the toxins, protecting and
restoring normal cellular functions while promoting elimination. The major
constituents of the volatile oils are: myrcene (1.71%), d-linalool (52.26%),
citronellol (4.64%), geraniol (9.29%), safrole (2.67%), aterpinyl acetate



(1.07%) and geraniol acetate. A typical dose is orally 6-15 drops 1/2 hr.
before or 1 hr. after meals 2x/day. For 5 days. 2 day rest and continue. Or
Apply 1/4 to 1/2 dropper on wrists, joints, or affected areas twice a day.
Vitamin B6: needed in the metabolic process that converts methionine to
cysteine and then into glutathione. B6 is capable of reducing and controlling
the swelling and pain associated with the routine tissue and bone trauma
resulting from normal dental operative procedures. You can also use
Pyridoxal-5-phosphate (P5P), the active form that B6 is converted to in the
body. Vitamin B1: is capable of reducing pain that may be associated with
routine dental operative procedures. B, is one of two vitamins containing
sulfur, the other is Biotin.

Magnesium: Magnesium availablility is essential for the proper functinoing
of our immune system as well as hundreds of enzyme systems critical to
human health. Organically amino acid-bound ones are more easily
absorbed and are less irritating to the gastrointestinal tract as well.
Activated charcoal: taken immediately with chlorella, 15 minutes before
drilling/chunking out amalgam, will bind any swallowed mercury and also
prevent recirculation in the liver.

Refrain from taking any supplements that contain iron and copper. Mercury
amalgam removal alone does not put an end to the mercury poisoning. The
mercury which leached from the fillings in the mouth is stored in cells
throughout the body and continues to exert its damaging influence. It is not
unusual to see patients who have had their amalgam fillings removed and
replaced ten to fifteen years prior to testing still having elevated levels of

mercury in the body. Once mercury toxicity has been demonstrated, by
tests such as high electrogalvanism, high mercury vapor emissions, and/or
high mercury body burden, mercury amalgam removal and replacement
with alternate, non-toxic materials is the recommended step. Botanical
substances to assist in removing the mercury include cilantro and chlorella
which are particularly effective.

Sweating
The skin is the body's largest detoxiification organs and sweating can help
draw mercury from the body. Saunas are a useful adjunct to safe mercury
removal because they induce copious sweating. Initiate sweating and
increased circulation by exercising 20 minutes three times a day on a
rebounder (mini trampoline). Immediately following the exercise, sit in a



sauna or under infrared lights (infrared sauna) for up to 30 minutes, then
take a cool shower. The temperature from a "low heat" sauna should be
between 140 to 180 degrees F. in contrast to the 200 to 210 degree F. for a
non-therapeutic standard sauna. The sauna may be followed by a plunge
into a bath or under a shower whose temperature is 65 degrees F. Over a
period of three to four days, increase your time in the sauna to a total of up

to two hours, divided into 30-minute periods with a short cooling-off period
in between. It's important to shower and towel dry because the removal of
sweat prevents reabsorption of toxins. While doing the sauna program,
consume adequate amounts of water to avoid dehydration. this is a
minimum of two quarts before and after entering the sauna. Replace your
electrolytes lost to perspiration with grape or prune juice and drink
vegetable juices to replace calcium and magnesium lost through the skin.
Oral Metal Chelation

NDF (Nanocolloidal Detox Factors)

Based on the results of comparative 24 hour urine samples analyzed by an
independent clinic and lab, a person can safely excrete up to 920% (9.2
times) more heavy metals per month taking NDF daily as compared to
doing one DMPS intravenous injection per month. This greatly shortens the
time required to achieve detoxification, an average toxic adult person
requiring a maximum dose of 2 mis. twice a day for a period of about two
months. NDF also removes other toxins from the system.
The predominant route of excretion is via the urine, thus accelerating the
excretion rate of the mobilized metals as compared to the fecal route,
decreasing the possibility of enzyme and leaky gut mediated resorption
through the bowel, and decreasing the burden on the liver. The majority of
the metals to be mobilized and eliminated per dose are quickly detectable in
the first urination following the dose. Fecal Element studies show an
average of 38.4% reduction in fecal metals following 5 days at maximum
dosage while urine levels remain elevated. Individual pathways of
elimination have been noted. Independent real time digital EEG studies
show a beneficial effect on the electrical activity of the brain, specifically

raises the heavy metal suppressed beta waves to normal levels (from within
to 113 minutes post ingestion and lasting at least 4 hours) with a
concurrent dramatic increase in the urinary excretion of heavy metals and
patient reports of subjective improvement. This proves that no "healing
crisis" is required during heavy metal detox while using NDF.

Ingredients:



2 milliliters (2 droppers full or 52 drops) contain:
50 mgs. - Nanocolloidal cell wall decimated Chlorella Pyrenoidosa
.12 mis. - Nanocolloidal Cilantro
mgs. - Nanocolloidal *PolyFlor
75 mgs/liter nanocolloidal Silica
Grain neutral spirits 18% as a preservative
*PolyFlor microorganisms include: 12 strains of lactobacillus (including
casei, acidophilus, salivarius, bulgaricus, sporogones and plantarum), 3
strains bifidobacterium including longum and bifidum, streptococcus
thermophilus, and b. laterosporus.

Why "Nanonize" the Ingredients?

Chlorella is known in mining to bind heavy metals to its cell wall. Yet many
people have taken Chlorella with no benefit. The reasons are that all of the
available chlorella is not really "cell wall broken" and that most of it is
already contaminated with heavy metals. Most of the cell walls are in tact,
but the individual diatoms are tightly clustered in groups of about 500 units
each. This is very difficult to digest and may explain why some people get
gastro intestinal distress when taking normal chlorella but not with NDF.
Nanocolloidal cell wall decimated chlorella has never been available so far!
In addition to binding to heavy metals, Chlorella has other beneficial
effects,
augmented by putting it through this process, including: increased
elimination of toxins, growth hormone regulation, a powerful nutritive impact
and protection from radiation.

Why does it work?

The following is essential to the understanding of this supplement: The
ingredients are in a nanocolloidal form. There is at least a 500-fold increase

in available surface area and a dramatically reduced particle size, thus
rendering each ingredient more bioavailable and effective. That means the
effective bioavailable dose is roughly one five hundredth of the dose
required compared to using a dose of the original ingredient. This is why 50
milligrams of nanonized chlorella achieves what 25 grams of normal
chlorella cannot. Most toxin-burdened people have compromised
assimilation and utilization and can't benefit from macromolecules.
In the past, Chlorella was only known to mobilize a small amount of heavy
metals via the bowel. In NDF, because it is nanonized, "molecular
components digested off the nano particles can be absorbed across the GI
wall into the bloodstream and have a possibility to enter the brain



depending on the molecule" - a possible explanation of why it can facilitate
elimination via the urine.
PolyFlor contains fulvic acid. This could be the underlying reason why
healthy bowel flora is so vital to good health. However, just taking a flora
supplement will not provide heavy metal detox of the same magnitude as
NDF (www.fulvic.com).
The major health benefits of both live and cell wall broken beneficial
bacteria are described by recent clinical research in The Handbook of
Probiotics. Lee, Nomoto, Salminen, and Gorbach. Pub. Wiley & Sons, Inc.
'99. Unfortunately, once the amalgams are put into the teeth, or the toxic
body burden becomes too great, or if a person only consumes processed
and pesticide grown foods, these powerful allies no longer stand a chance
of sharing their healing benefits with us.

Duration of Therapy

So far, only how much metal is being excreted can be measured, not the
total body burden, so it is impossible to exactly predict the duration or cost

of therapy. We do know that there is a linear relationship between the
volume of the dose and the amount of excreted metals. Therefore, the more
they can take, the quicker the detox will be. However, it is preferable to
maintain the dose at the level that the patient continuously reports
subjective improvement as a "healing crisis" is not required to effectively
remove the heavy metals with NDF.

Cost Effectiveness / Compared to DMPS

It was recently determined by an independent, comparative 24-hour urine
tests conducted by Dr. J. Wright via Doctors Data that a single, 2-dropper
dose of NDF pulled out 20% as much metals as an IV dose of DMPS on the
same patient. Since NDF can be taken daily, and DMPS only once a month
(per the protocol presented by Drs. Klinghart and Mercola), this means that
up to 920% (9.2 times) more metal can be excreted per month using 2
droppers of NDF twice a day (maximum dose) without the side effects and
mineral deficiencies associated with DMPS. Since there is a linear
relationship between amount of the dose and percentage increase in
excreted metals, 6 drops twice a day would take out about 107%, or roughly
the same amount of metals per month as DMPS, making NDF very cost
effective, especially when you consider that very little additional
supplementation is required while using NDF. Suggested retail is now $150
for a one-month supply, equal to the cost and efficiency of one DMPS IV
push.



Rectal Chelation

New Delivery Method for Chelation Therapy
The newest, easiest, most convenient and efficacious technique for
detoxifying heavy metals out of the body is by means of rectal chelation
therapy. The method is to self-apply Detoxamin, a patented, trademarked
and registered over-the-counter suppository. People exhibiting toxic metal
burdens now are able to chelate themselves while sleeping by use of this
non-prescription chelator. Merely insert the firm gelatin pill into the
rectum,
go to sleep, and awake in the morning partially detoxified. Repeat the
procedure until testing show that there is no more metal poison remaining in
the body. With this suppository method, the main obstacle to intravenous
EDTA chelation therapy has been eliminated. Rather than spending three
or more hours per infusion session in a clinic, hooked to an IV, you may
take less than a minute to insert the Detoxamin suppository at home before
bedtime. Since many people cringe at the thought of getting stuck with a
needle for twenty or more such IV treatments, use of a suppository
eliminates this psychologically stressful and time-consuming obstacle.
Rectal administration is less invasive, in no way uncomfortable, and
generally greatly preferred over IV treatments.
Taking 3-5 suppositories over a 30-day period. This is medically equal to
approximately 2-EDTA IV treatments. When on Detoxamin maintenance
one box of Detoxamin lasts 6 to 10 months. Taken every night for 90 days
or every other night for 180 days provides the medical equivalence of
approximately 30 IV Chelation treatments.
Rectal chelation therapy does the job of detoxifying in a low-cost way to
effuse EDTA through the bowel's walls and into your blood stream to clean
toxic metals from all body cells. Detoxamin has a time-release mechanism
that allows the EDTA to absorb through the colon wall over an eighty-
minute period while you sleep. Almost all the blood from the rectum makes
its way to the superior hemorrhoidal veins, a tributary of the portal system,
so that absorption through the rectal wall carries the EDTA in Detoxamin to
the portal vein.
The lower and middle hemorrhoidal veins bypass the liver-and do not
undergo first pass metabolism. This means that the EDTA in Detoxamin
goes directly to the organs of your body without being filtered through the
liver first. Because of this, the EDTA contained in Detoxamin is very
productive. Detoxamin also introduces EDTA directly into the systemic
circulation, efficiently bypassing the portal circulation and the liver
metabolism on the first pass. Rectal absorption may also occur through the



lymphatic system and, in some cases, largely through the blood via the
vena cava.
Detoxamin offers many advantages both over the expensive intravenous
method of EDTA chelation. With the use of needles via the intravenous
method, and risk of AIDS and other communicable blood-borne diseases,
Detoxamin is becoming the logical choice over I.V. EDTA chelation and the
poorly absorbed oral EDTA. The rectum has a more neutral pH and is not
as acidic as the stomach, which makes this area much better for EDTA
absorption because it is not buffered and has a neutral pH, unlike the
stomach. It also has very little enzymatic activity, thus enzymatic
degradation does not occur. The rectal mucosa (rectum) is much more
capable than the gastric mucosa (stomach) of tolerating various drug-
related irritations. This is why patients who can't tolerate oral pain
medication are given the same medication in suppository form. In fact,
absorption with any oral EDTA tablet is so low that 135 (500mg) oral EDTA
tablets are equal to just 5 Detoxamin suppositories.
Detoxamin removes most harmful toxins from the body, safely and
effectively. Detoxamin is taken at night prior to bedtime, each Detoxamin
suppository contains 750mg of Calcium-disodium EDTA, and is made in a
cocoa-butter base (melts on body contact), which is very therapeutic for the
rectal mucosa and the colon wall. The Ca-sodium form is able to bond
(chelate) effectively because it does not lower the blood pH to a level that
would prohibit the bonding action. The Ca added to the salt is important in
this mode of administration as it buffers the acidic quality of the active
ingredient keeping the suppository from being abrasive to the mucous
membrane of the rectum area. Ca-disodium EDTA has both a scientific
justification for therapeutic effectiveness as well as a clinical history of
effectiveness.
The Calcium EDTA in Detoxamin has an extra chemical bond compared to
the older Disodium EDTA. This gives Detoxamin EDTA an affinity for
Mercury. Mercury is also excreted from the body through the feces and,
because Detoxamin utilizes the colon wall for EDTA assimilation; it is a
powerful Mercury chelator.
Metal Removing Nutrients
Calcium & Vitamin C: Just as lead will displace calcium, calcium is an
excellent nutrient to utilize for displacing mercury and lead. Utilizing a
combination of minerals, such as magnesium and calcium, is even more
effective in clearing metals from the body. Increasing vitamin C intake is a
reasonable cost-effective way to control toxic metal levels in the population.

Several studies implicate lead in causing cavities, and at least one study




suggests that almost 3 million cavities in children result from lead. Vitamin
C and Calcium supplementation are recommended for protection.
Chlorophyll: chlorophyll binds to heavy metals very well. In fact, it is
imperative to choose a reputable source for your chlorophyll, which screens
for toxins and heavy metals; or you may be getting more than you want. A
good source is juiced raw, organic greens.
Fiber: Fiber, such as oat bran and apple pectin, will bind to metals and help
draw them out of the body. Montmorillinite clay also binds extremely well to
toxins and metals for clearance. Fiber such as red beet root fiber is high in
proanthocyanidins and antioxidants and facilitates clearance of metals
through the liver.
Lipoic Acid: Lipoic acid is a potent antioxidant and has a high affinity for
binding to metals. This makes it an excellent choice as a supplement to
bind and clear mercury and lead from the system. It is best utilized in
combination with conjugating nutrients.
Minerals: A mineral-rich diet acts as a chelating agent. Many minerals will
chelate metals, including calcium, magnesium, zinc and selenium. Mercury
interferes with some functions of selenium, including its powerful antioxidant

function and its ability to bind to metals. A good source of bioavailable
minerals is from raw sea vegetables and grass juices from wheat, barley,
alfalfa, kamut, etc.
Milk Thistle (silybum marianum): Milk thistle is a renowned liver herb, and
supports this major detoxification organ. Milk thistle contains silymarin, a
bioflavonoid that is a very potent remedy for the liver. Silymarin inhibits
free
radical damage; free radicals have an adverse effect on the detoxification
enzymes of the liver cytochrome P450 system, while silymarin protects
those enzymes. Glutathione is destroyed by lead. Silymarin not only
prevents the depletion of GSH (glutathione), it even increased this liver-
detoxifying enzyme. A sulfur pathway in the liver detoxes lead, and milk
thistle helps to boost liver function.
Molybdenum: Large amounts of exogenous sulfur (from outside the body)
will usurp the body's stores of molybdenum to metabolize it. An easier
solution is to use the nutrients which will facilitate the homocysteine
pathway. Homocysteine is a toxic substance, however the pathway itself,
when properly supported, is essential for a host of metabolic functions.
When the pathway is facilitated, sulfur is generated as a natural by-product
at the end (molybdenum changes the toxic sulfite molecule to the much-
needed sulfate). Vitamins B12, B6 and folic acid, along with trimethylglycine
and dimethylglycine recycle homocysteine to methionine, and allow for
Sam-e to methylate phosphatidylserine, an important brain nutrient. Usually




the people who are the most deficient in sulfur will be the most sensitive to
metal toxicity and vice versa.
Parotid Glandular: Parotid glandular is believed to accelerate the clearance
of chemicals/heavy metals from tissues. It is best utilized in combination
with detoxification nutrients that will pull the metals out of the body by
detox
pathways such as the bowel, kidney, lymph, lungs, blood, skin, and liver.
Sulfur: Lead, mercury and cadmium steal sulfur from important proteins,
which could be enzymes, hormones, or cell receptors. Conversely, sulfur is
needed in the liver detox pathway to hook onto these metals and clear them
from the body. So, lead, mercury and cadmium depletes sulfur, the very
nutrient needed to detox the metal overload. A depletion of sulfur will also
adversely affect joint connective tissue growth, since sulfur is an essential
precursor to the building blocks of cartilage, namely glucosamine sulfate,
chondroitin sulfate, and hyaluronic acid. Good sources are egg yolks, garlic,
kelp, kale, turnip, raspberries, onions, cabbage, and mustard.
Zinc: Zinc and copper get displaced from metallothionine, the protein that
binds and carries them. This destroys many of the zinc-dependent
enzymes. Zinc is important for proper functioning in a host of major
metabolic pathways; it is a component of over 90 metalloenzymes in the
body. Lead has always been known as a neurotoxin, with the brain being
particularly susceptible to attack. Lethargy is a common symptom of lead
toxicity; lead inactivates the zinc-dependent enzymes of the Kreb's cycle,
which produces our energy. Zinc is also a part of the antioxidant enzyme,
Zn-SOD, which fights superoxide radicals. Symptoms of lead toxicity are
similar to zinc deficiency symptoms because lead can bring on a zinc
deficiency. Zinc deficiency has been implicated in a wide variety of
neuropsychiatric disorders, including dyslexia, epilepsy, mental depression,
and attention deficit disorder. The symptoms of lead toxicity are similar to
zinc deficiency because the lead destroys the zinc-dependent enzymes.

Natural Healing
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We are also working on 1) Isolating the most effective enzymes from ant's
stomachs (also
bis-peptides and other amino acids - eg. to fight disease...in gene therapy,
and all genetic
engineering, such as making stem cells proliferate longer, by altering their
genes) and any
and all simpler creations, 2) then duplicating their molecular structure using
(atom)
teleportation technology, 3) then eventually altering the pattern of the
molecule feed stock to
produce new molecules (designer), from enzymes to drugs for the pharmacuetical
industries
and new materials such as not just diamonds, but nano particles that are safe
for human
biology, more strong, more flexible, more dazzling, more sensitive sensors
(eg. to sound
vibrations), more heat resistant, any and all materials new qualities.




If we use a quantum computer/Artificial Intelligence Node Network job
sharing/Huge Main
Frame we may be able to mass assemble these duplicate produced via
teleportation
technology economically.

We are also considering mixing stem cells with specialized cells in tissue
culture as well as
separate and/or together in delivery/administering as well as grafting from
for example tissue
culture.

Converting stem cells to organ connective tissue using a scaffold whose
dimensions are
standard/default/similar in proportions for a person (possibly a donor
dissected and detergent
removed only connective tissue left) and then measuring the patient's actual
dimensions (eg.
by PET, CAT scan, x-ray) (measuring the usual and important areas) of
differences in size
shape (eg. veins and artery and ventricle tissue).

We could also use to replace fat, lipo suction, then replaced with high
metabolic cells and/or
stem cells.

We could also use the stem cells and/or specialized cells for breast
augmentation and penis
(and/or using scaffolding to grow a new larger penis from stem cells)
augmentation. Perhaps
one way is for the penis augmentation to work is to study the specialized
cells relative to
thickness/diameter/shape of the penis. Where is the sensation nerve
connections located in
the cells, how much volume increasing cells are spread out where, dose the
patient have the
blood supply to support a fully erect penis. To test sensations, we could test
on animals (eg.
rats), what type of specialized cells provide sensations (how much is muscle
memory) can
stem cells be used instead (directly) in the theory that the stem cells will
move change into
the specialized cells that surround it. Additionally can too much sensation
cells cause pain,
we can tell if the rats avoids errections (even under viagra) and/or ignores a
female in heat
(pheromones). Furthermore, with the new enlarged penis, can the can (and how
can we
make the) the blood flow increase if needed? Furthermore we could study pigs
which are
known to have 1/2 hour orgasms (are they male or female), their cellular make
up -
scaffolding, connective tissue, ability to remain erect, perhaps it's in the
continuous supply of
blood or their mind to nerve muscle memory or their internal plumbing such as
larger supply
of orgasmic fluid relative to pump rate, or perhaps it's their slow fat (slown
down by fat tissue)
metabolism and/or types of difference in nerve cells and/or brain activity
during orgasm.
Perhaps we could use neural electrodes and/or mind sounds and/or direct muscle
patch to
simulate mimic brain activity during orgasm creating artificial orgasms with
no end in time).
We might also mimic (using neural electrodes and/or mind sounds and/or direct
muscle
patch) for the brain activity that causes and maintains errection and for
females faster sexual
escalation to orgasm. The above might be the cures for any and all sexual
dysfunctional
diseases (eg. impotence, early/delayed ejaculation, as well as size and
hardness, and
frigidity). Another option for for example opposite sexed mates is 1. a touch
sensitive
artificial penis - changeable sizes - diameter and lengths (for men and for
robots with
emotional artificial intelligence) that is controlled via two way signals (to
read the sensations
in both the man and also the female are being felt as the penis and vagina
interact...). We do
this by using neural electrodes and/or mind sounds and/or direct muscle patch
for men and
robots to satisfy women as well as the man (via signals back to the man's
brain/mind that
mimic and/or duplicates the feeling of orgasm in the mind - we could even
continue this
feeling for both men and women all day long) himself at the same time. 2. for
women, we
could use the neural electrodes and/or mind sounds and/or muscle patches that
induce
and/or work in synchronicity to push (flood rhythm, sensations) the woman to
orgasm and/or
mimic the feeling of orgasm to the brain/mind all day long. We could create a
muscle patch




electrode(s) (as well as vibrating) that are attached the artificial penis
that will stimulate the
woman's g-spot and/or any and all sensations cause the woman to orgasm - in
addition to
regular foreplay. It is my hope that eventually people could induce orgasms
artificially
without dwelling on dirty thoughts or pornography. Or at least this invention
will enhance the
sex lives of people who are sexually dysfunctional. We could stem cells that
are either
differentiated into the specialized corpus cavernosal smooth muscle, the
endothelium of the
sinusoids, blood vessels and the smooth muscle cells in the penis that produce
transmitters
and modulators...all tissue cultured in the laboratory, then either 1) we
(multiple micro) inject
the differentiated cells in to the places that we want to bulk up, 2) we
(multiple micro) inject
stem cells directly into these specialized areas of the penis in the hope that
the stem cells will
migrate and differentiate into the localized cells via cell to cell
signalling, 3) we can create a
whole new larger and longer penis that can be grafted on the patient to
replace the old one.
We think exercises and more arteries/and larger arteries capacity (eg. larger
inside scaffold
that the arterial cells grow around the hollow hole) and inversely the less
and smaller inside
hollow capacity of venous outflow, the better for a more erect rather than
flaccid penis. To
enhance conversion of stem cells to specialize into endothelial growth, we can
use 1)
mechanical forces, (fluid shear stress and cyclic stretch - pulse and
perodicity) eg. gels that
emulate the pressure conditions of the penis at erect and rest, 2) endothelial
growth factor
VEGF-C, EG-VEGF, Phospho-VGFR2 (Tyr1214), VEGF121,VEGF-A, VEGF-C, VEGFD,
VEGF-R1, VEGF-R1/Flt-1 (17a.aC term), VEGFR2(Ab-1214) Antibody, VEGF-R2/Flk-
1/KDR
(188a.a C term), VEGFR2/KDR, VEGF-R3/Flt-4 (20a.a C term); TGF-B2, KIAA0101,
CARP
and syntenin, CXCR-4, TGM-2, LIM-domain protein and claudin-5; we could also
use these
growth factors to increase proliferation of stem cells (VEGF-C) for
proliferation, migration,
through selective binding and phosphorylation of tyrosine kinase, growth
factor receptors (eg.
kdr, flt-4 and flt-1). 3) Another way to morph the stem cells is to create a
medium or serum
derived from the specific cell type of large animal (fibroblasts), such as a
cow, horse and/or
pig for the final morphing stage of the corresponding to the same cell
type/portion of penis on
humans - the assumption is that the human stem cells will mature into the same
cells as
those surrounding it.

The idea of grafting may also be possible, but we would need to know the
sensation
implications in terms of pattern of nerve cells and their necessary
connections to the original
nerve network below. Grafting may make the sexual sensations numb if not
embedded with
the right pattern of nerves and furthermore rough sex may tear the graft.

We could also mix injections with grafting.

We could use micro multiple syringes to deliver (connective tissue (and/or
stem cells) -
which can be injected into spread out (together in conjunction with training)
areas of the gut -
to strengthen for example medicine bali training in boxing and/or iron shirt
qi gong).

Fast twitch muscles and/or their progenitor stem cells using micro multiple
injections could be
used to deliver to strategic locations (areas) that correlate to
shape/relative sizes or wanted
body muscle proportions. The patient can exercise to make the muscles grow and
perform.
This muscle augmentation can also be used for paralysed people in conjunction
with stem
cells and neural cells to the spine and the finger/palm therapy Gerard
developed especially in
cases where the muscles have deteriorated in size and strength.

Fast twitch muscles and/or their progenitor stem cells using micro multiple
injections could be
used to deliver to strategic locations (areas) that correlate to
shape/relative sizes or wanted
body muscle proportions. The patient can exercise to make the muscles grow and
perform.




This muscle augmentation can also be used for paralysed people in conjunction
with stem
cells and neural cells to the spine and the finger/palm therapy Gerard
developed especially in
cases where the muscles have deteriorated in size and strength.

Additionally to create a cloned human being; we take a fertilized that has
formed a
blastocyst, remove the fertilized stem cells, inject an adult patient/desired
stem cells (to take
the place of the stem cells of the fertilized egg/sperm) that has come from
tissue culture - eg.
earlier cloned stem cells (or try somatic adult stem cells - there is the
caveat that somatic
cells may not divide), then we attach the blastocyst to a female's uterus and
grow the fetus to
a cloned baby.

Additionally to create a cloned human being; we take a fertilized that has
formed a
blastocyst, remove the fertilized stem cells (via through the outside of the
same site that the
this inner cell mass (ICM) is attached to on the blastocyst - Trophectoderm
outer blastocyst
wall(TE)), we use the same site (by using the site where the ICM is stuck to
the blastocyst
TE we avoid tearing or implanting the stem cells into the wrong side (of the
Mesoderm) that
separates the inner cell mass from the blastocoel - fluid cavity) to inject an
adult
patient/desired stem cells (to take the place of the stem cells of the
fertilized egg/sperm) that
has come from tissue culture - eg. earlier cloned stem cells - possibly
electro fusing, laser
fusing and/or heat shock (or try somatic adult stem cells - there is the
caveat that somatic
cells may not divide), then we attach the blastocyst to a female's uterus and
grow the fetus to
a cloned baby. To make this work we may need create (stuff the new stem cells)
enough to
create the same pressure conditions as the previous fertilized egg stem cells
separated by
the mesoderm.

To collect single adult female stem cells for use in replacing stem cells in
the blastocyst the
following process can be used:

Stem Cells from female eggs can be obtained as follows:
1. Treat the eggs with oncogene c-Myc.
2. Hyper ovulate (use of fertility drugs)
3. Harvest eggs.
4. Mature them in CMRL-1066 media (Sigma), FCS (HyClone), pregnant mare
serum (Sigma), human chrionic gonadotropin (Sigma), penicillin, streptomycin.
5. The eggs are incubated in CO2, inonomycin, then in 6-dimethyliaminopurine.
6. The stem cells are then removed and further media are similar to the media
above (stem cell tissue culture - especially proliferation; since before we
can treat
patients we need sizable inventory of compatible stem cells first).
7. We could use electroshock, heat shock to get the stem cells to start
dividing.
8. We could also use the above maturation cycle and incubation to proliferate
stem
cells produced by the traditional method (eg. Oocyte removed nucleus, micro;
laser; electro shock and replaced with somatic DNA).
9. Alternatively we could use primordial germ cells from an adult male and use
the
above maturation cycle and incubation to proliferate stem cells.
10. Another alternative is to use a batch of stem cells (from any of the above

methods) that have stopped proliferating and either treat with the maturation
cycle, then incubate to jump start the proliferation cycle again.
11. Or we could use a batch of stem cells (from any of the above methods) and
microinject, laser electroshock the DNA from the stem cells (that have stopped

proliferating) into a fresh Oocyte (with the nucleus removed).




12. We could also alternate between 10) and 11) or test which one is the most
prolific
and which one is best for maintaining the integrity of the DNA - from mutating
or
degenerating.

We might also use (perhaps electron/nano/quantum) video, microscopic thermal
heat
sensors to view the innards (internal bodies and fluids) within the cell.

We are also considering growth factors and gene therapy combinations of the
following:
FGF-1 and FGF-2; and at least one of the following: VEGF, VEGFA, VEGFB, PLGF,
VEGF121, VEGF145, VEGF165; VEGF189, VEGF206, FGF-1, FGF-2, including the
following gene therapy is AD5(FGF4) or VEGF 165 plasmid DNA to extend stem
cell
proliferation.

We also suggest that if you cut the nerves to a salamander's arm the
salamander may
regrow an extra limb. Based on this theory we find that the salamander's
nervous system
senses that the limb is missing and regrow's the third limb.

Perhaps humans could regrow limbs and organs by growing using a blastema (stem
cells)
with the Human HOX genes and pressure that is found in that area of the body
as well as cell
signalling, with new doses of stem cells to be added as the stem cells migrate
and form the
different stages of embryonic development of the limb and/or organ.

The other option is to inject the stem cells together with activated HOX genes
regularly
(perhaps by Intra Venous drip GP; GV) and in time before scarring even use
topical and/or
abrasive dermal removal of scar tissue, even shave of the overgrown tissue in
preparation
for the mixture of stem cells with activated HOX gene to create a human
blastema as well as
cell signalling. The stem cells with activated HOX genes can be added directly
to the (eg.
removed area - limb) as the progress of migration and distal tip, stage by
stage
embryogenesis converts and/or causes the stem cells to produce the limb/organs
that is just
in time method, so there is no excess or not enough stem cells with the
activated HOX genes
to cause unwanted growth or in the case of not enough administration scar
tissue may form
or the regeneration may be instructed to stop due to lack of stem cells with
activated HOX
genes by whatever the body's recognition is, ... and also where the best place
to administer
the concoction, and amounts...

Furthermore through any and/or all gene therapies we could activate the HOX
genes in stem
cells to further proliferate them.

We are considering injecting homeodomain proteins in serum mixed with/or
separately, stem
cells, (differentiated cells appropriate for that part of the body) eg.
dendrites, neural cells and
neutrophins, into brain ill people (eg. parkinson's disease, paralysis - brain
damage,
alzheimer's, Mad Cow Disease...). Furthermore we could saw off part of the
skull and add
homeodomain proteins in serum mixed with/or separately, stem cells,
(differentiated cells
appropriate for that part of the body) eg. dendrites, neural cells and
neutrophins (using
previously artificially grown blood brain barrier tissue (and/or donor) -
and/or blood brain
barrier from a primate. We could fuse it to a pourous bone matrix that has
blood and or
artificial blood to exchange waste for nutrients needed and can be temporarily
removed for
cleaning - the purpose is to see what happens to the mental abilities of the
subject.

This technologly of Homeodomain in the hope it will be absorbed by localized
cells in
conjunction with precusor stem cells and cells differentiated for that part of
the body.




The first step is to study what and under what conditions (eg. pressure, salt
concentrations)
substances are facilitated from which side of the blood brain barriers to
which side.

Humans generally contain homeobox genes in four clusters:
Image

There is also a"distal-less homeobox" family: DLX1, DLX2, DLX3, DLX4, DLX,
and DLX6.
"HESX homeobox 1" is also known as HESX1.
Short stature homeobox gene is also known as SHOX
3%

1%
3%
We are trying to revive dinosaurs' and extinct/or and nearly extinct species.
We plan to use
the Hox genes in different ways.

We can add the hox genes (into the nuclear membrane and then either from micro
inject
and/or retro virus then electroshock, laser fuse, heat shock and fill their
cytoplasm (GP 25%)
and the hox and transcription factors with the building blocks at the ovum
and/or zygote
stage of development. And we could remove a fetus from an egg and mix the egg
white of
another egg with evenly spread hox building blocks (plus hox genes) and hox
transcription
factors then we could place the fetus in anther egg, with egg white mixed hox
material and
egg white even in an artificial shell matrix to incubate. We need to study
whether hox should
be introduced at ovum, zygote, or at fetal stage - before sometime during
growth, as growth
is slowing down and or when and/or after growth has stopped. We could of
course study
how the genes (activated versus inactivated, what is the sources of and which
genes are at
the top of the pecking order causing which other obviously essential genes
(for continued




growth into dinosaurs) to silence at the stage of fetal development where the
fetus of a
chicken diverge from the fetus of an alligator. What if any environmental
(versus pre-past
historical gene mutation programming coming evolution and natural selection)
triggers signal
these genes to activate, when is a an embryo of a chicken's teeth too large
and what tells
which genes in the chicken genome to recede the teeth ... Evolutionarily
speaking it is
possible that the stage of growth (eg. incubation stage) of the fetus
determines at some
stage in evolution oc the chicken species, mutated chickens fetuses that lose
their teeth past
a certain stage in the fetal development dominated (survived in their habitat)
versus those
that grew teeth beyond that point. So by using hox gene we expect to carry the
chicken to
continue teeth growth all the way to hatching (continuing the growth of its
body parts through
the stages that such body parts would in current evolutionary status of
chickens to day to
dinosaurs derived from the continued growth).

Alternatively and additionally we could protoplast fuse chickens, emus,
ostriches, alligators
and crocodiles.

Finally we could study chickens, emus and ostriches to see which genes are
turned/turned
off, activated/deactivated, silenced other than hox genes whereby chickens
with teeth have
certain gene drivers, that are changed from no teeth, which genes
control/regulate and are if
any also responsible for the suppression of dinosaur hatchlings.

To genetic engineer dinosaurs we use chickens, emus, ostriches, alligators and
crocodiles.
The vertebrate homeotic complex comprises four distinct Hox gene clusters (Hox
A, B, C, D)
that are organized into thirteen homology (or paralogue) groups.


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CA 02629555 2008-05-14

We could process bio crops such as Jatropha, corn, sugar cane, coconuts and
oil palm, as
well as any and all organic wastes (including manures, sewage)...

1. Chemical solvents: hexane, benzene...
2. Soxhlet: chemical solvents, methodology involves repeated washing and/or
percolation using hexane and/or petroleum ether (involving reflux in specified
glassware).
3. Enzymes (best bred/cloned/synthesized): enzymes break down the cell walls
of the
bio crops, involving water as solvent assisting in fractionation.
4. Expression (Expeller Press): Mechanical pressing and chemical solvents.
5. Osmotic Shock: fast reduction in osmotic pressure causing the cells to
implode/explode, to expel cellular contents including oil.
6. Supercritical fluid: C02, is liquefied under pressure and heated which acts
as a
solvent that is both in a liquid and a gaseous state.
7. Ultrasonic-assist: sonochemistry, using an ultrasonic reactor, ultrasonic
waves make
captivation bubbles in a solvent material, the bubbles rupture near the cell
walis, it
makes shock waves of liquid jets making the cell walls to break down to expel
their
contents within a solvent.
8. We plan to use our diamond technology for manufacturing larger diamonds,
used in
conjunction with the pressure/heat to press to crush and breakdown bio crops
to
release their oils (GP 10%), and any and all waste including garbage, eg.
plastic,
rubber, and oil/tar sands (into slag and gasses) we could also combine the use
of the
pressure/heat smelt metals (steel, aluminium..) heat salt water and use the
steam to
turn turbines producing energy and then condensating into fresh water, the
heat can
be powered by mirrors or Tall Gravity to Electricity Invention *underground
patented
by Gerard Voon) and heat to vibrations to energy technology.
We could do any and all combinations of the 1 to 8 processes above to process
for any and
all bio-crops and any and all garbage as well as in tandem, we could sequence
the
processes. (DLD 40% Asia)

We could harvest (via from animals and cells tissue to artificial synthesis) 1-
alpha-
hydroxylase calcidiol to calcitriol, to produce 1,25-dihydroxyvitamin D that
bond to Vitamin D
Receptor and retenoid-x receptor (RXR).

We are using the products of VDRE vitamin D response elements namely
catheiicidin and
defensin beta 2 (Peptides) used as an antibacterial/antiviral/antifungi
agents. Including all of
the above uses, that are interchangeable with chitosan eg. cleanser -
detergent, carpet
powder, soap bars, liquid soaps, disinfectants, tooth paste, mouth wash, final
stage to clean
up industries that use strong microbes (eg. genetically engineered extra tough
microbes), -
we are also testing these two substances effectiveness against toxins and
heavy metals (eg.
binding...).

We plan to use 1,25D as a treatment to cause immune cells take away the cell
to cell
signalling via secreted cytokines and other factors that cause major
inflammations. Diseases
that can be treated include, but are not limited to, peripheral - chronic
inflammation-related
diseases, for example: chronic inflammation; thrombosis; atherosclerosis;
restenosis; chronic
venous insufficiency; recurrent bacterial infections; sepsis; cutaneous
infections; renal
disease; glomerulonephritis; fibrotic lung disease; allergic disease; IBS;
rheumatorid arthritis
and acute bronchiolitis. Central nervous system-macroglia and microglia
related diseases, for
example: neurodegenerative diseases; Alzheimer's disease; Multiple sclerosis;
Parkinson's
disease; neuroinflammation; HIV-associated neurological diseases; HIV-
associated


CA 02629555 2008-05-14

dementia; CNS bacterial infections; brain Toxoplasma gondii; Acanthamoeba
infections;
Listeria infections; prion diseases; subacute spongiform encephalopathies and
macular
degeneration may also be treated.

We also plan to deactivate, remove, the GADD45x-is gene to try to augment our
stem cells
to more proliferation, then restore its function once the cells are ready for
use.


CA 02629555 2008-05-14

As well as the above creatures we could use the HOX treatments on (close
relatives of the
dinosaur precusor), eg. elephants to mammoths, tigers to sabre tooth tigers,
alligators/crocodiles, bats, any and all birds.

Any and all methods and techniques for Cloning Elephants, Narwhales, walruses
and any
and all expensive ivory, teeth, bones (whales), human, skulls, jaws, horns
(horns 5%).
We need to determine the precusor cells to the ivory (possible from the jaws,
and/or skill
and/or dentine surrounded on one side by pulp).

We need to know the signalling mode of these creatures that cause them to grow
the ivory
(eg. pressure missing, feedback molecules, hormones, substances and their
concentrations,
produced by the cells external surrounding cells and/or internally within the
cells eg. causing
the stem cells DNA to line up and the cell to split into two robust (cytoplasm
- organelles)
divisions)...

We could tissue culture their stem cells (especially from creatures with the
best quality ivory)
and find the gene(s) and/or chemicals that stimulate the stem cells to
specialize into ivory.
One method to enhance growth is to Gerard's patented stem cells recycling
method (and
perhaps isolation and further culturing the cells with the longest telomeres
into new colonies
possibly surrounded with dental pulp on one end)... We plan to grow any and
all teeth and
bones and horns and tusks possibly using precusor cells, (stem cells using my
patented cell
proliferation techniques possibly bone marrow and red blood cells and plasma).
We could
grow teeth, tusks, bones (including whale bones) by differentiating their stem
cells via
signalling and emulating the necessary factors (environment that) the cells
naturally grows in.
We will induce the bone cell differentiation via bone morphogenetic proteins
from large
numbers of stem cells and/or Germ cells and/or bone marrow derived from the
stem
cells/germ cells. We could use synthetic matrix/scaffold that are porous,
varying shapes and
sizes to seed new bone growth. Possibly using coral, ceramic and animal
collagen.

We could feed calcium and proteins to prompt and calls on cells such as
osteoblasts and
other cells to lay down new matrix that mineralizes and forms hydroxyapatite
and collagen.
By using a porous matrix/scaffolding we allow blood vessels (and/or synthetic
blood and/or
liquid/serum tissue culture medium) snake through the bone framework (and/or
applied
directly to control/manage growth of the teeth/bones/tusks/horns) carrying
many different
compounds that orchestrate bone remodeling, such as calcitrol (a form of
vitamin D-3),
caicitonin (a thyroid hormone that prevents bone resorption), parathyroid
hormone (which
works with calcitrol to regulate calcium and phosphate metabolism), and
prostagiandins (fatty
acids that perform hormonelike functions).

We could also use foam/polymer/co-polymer to contain the direct tissue culture
of bone
marrow (and/or precusor stem cells seeded for its own proliferation; also stem
cells into bone
marrow; and stem cells into red blood cells and plasma at the right time the
stem cells are
treated with signals, proteins, amino acids ... pressure, hormones, bone
morphogenetic
protein growth factors), surrounded with surrounded by live vascular tissue,
surrounded by
medium such as nutrients oxygenated pulsing with nutrients.

Additionally (we could use neurotrophin family includes nerve growth factor
(NGF), brain-
derived neurotrophic factor (BDNF), neurotrophin-1 (NT-1), neurotrophin-3 (NT-
3), and


CA 02629555 2008-05-14

neurotrophin-4 (NT-4) and/or apply with stem cells to apply to patients
suffering from spinal
cord injury).

REFERRENCE MATERIAL...
TAKEN FROM WIKIPEDIA:

List of Bone Morphogenetic Proteins

BMP Known functions Gene Locus
*BMP1 does not belong to the TGF-,6 family of proteins. It is a Chromosome: 8;
BMPI metalloprotease that acts on procollagen I, II, and Ill. It is Location:
8p21
involved in cartilage development.

Acts as a disulfide-linked homodimer and induces bone and Chromosome: 20;
BMP2 cartilage formation. It is a candidate as a retinoid mediator. Location:
20p12
Plays a key role in osteoblast differentiation.

Chromosome: 14;
BMP3 Induces bone formation
Location: 14p22
BMP4 Regulates the formation of teeth, limbs and bone from Chromosome: 14;
mesoderm. It also plays a role in fracture repair. Location: 14q22-q23

BMP5 Performs functions in cartilage development. Chromosome: 6;
Location: 6p12.1
Chromosome: 6;
BMP6 Plays a role in joint integrity in adults.
Location: 6p12.1
Plays a key role in osteoblast differentiation. It also induces Chromosome:
20;
BMP7 the production of SMAD1. Also key in renal development and Location:
20q13
repair.

BMP8a Involved in bone and cartilage development Chromosome: 1;
Location: 1 p35-p32


CA 02629555 2008-05-14

BMP8b Expressed in the hippocampus. Chromosome: 1;
Location: 1 p35-p32
BMP10 May play a role in the trabeculation of the embryonic heart. Chromosome:
2;
Location: 2p14
BMP15 May play a role in oocyte and follicular development. Chromosome: X;
Location: Xp11.2
hide]

v=d-e

Cell signaiing: TGF beta signaiing pathway
TGF beta family (TGF-01, TGF-02, TGF-03)
Bone morphogenetic proteins (BMP2, BMP3, BMP4,
BMP5, BMP6, BMP7, BMP8a, BMP8b, BMP10,
BMP15)
TGF beta superfamily of ligands Growth differentiation factors (GDF1, GDF2,
GDF3,
GDF5, GDF6, GDF7, Myostatin/GDF8, GDF9, GDF10,
GDF11, GDF15)
Other (Activin A and B/inhibin A and B, Anti-mullerian
hormone, Nodal)
TGFBR1: Activin type 1 receptors (ACVR1, ACVR1B,
ACVR1 C) - ACVRL1 - BMPR1 (BMPRIA - BMPR1 B)
TGF beta receptors TGFBR2: Activin type 2 receptors (ACVR2A, ACVR2B)
- AMHR2 - BMPR2
TGFBR3: betaglycan
TransducerslSMAD R-SMAD (SMAD1, SMAD2, SMAD3, SMAD5, SMAD9)
- I-SMAD (SMAD6, SMAD7) - SMAD4
Ligand Inhibitors Cerberus - Chordin - DAN - Decorin - Follistatin -
Gremlin - Lefty - LTBP1 - Noggin - THBS1
Coreceptors BAMBI - Cri to
Other SARA

Chemical & Engineering News
August 25, 1997


CA 02629555 2008-05-14
Other SARA

Chemical & Engineering News
August 25, 1997

Copyright 1997 by the American Chemical Society

I l- . . v19"~
BONING UP

At the human body shop, bioceramics or biopolymers combined with bone cells
and
growth factors are likely to lead the parts list

STEPHEN K. RITTER
C&EN Washington

A woman struck by a car in Florida loses 3 inches of her crushed lower leg and
could be left
to live with a drastic disability for the rest of her life. A Detroit teenager
with a marble-sized
divot in his thigh from the removal of a benign tumor could spend the rest of
his life being
overly cautious.

But the woman is now back to ballroom dancing, and the teenager is swimming
again and
plans to go skiing.

These dramatic tales of people who against great odds recovered from accidents
or diseases
to live reasonably normal lives are examples of the benefits reaped from
research that began
in the 1980s to characterize with increasing accuracy the content and
structure of human
bone. The success of this research now is resulting in new drugs and
treatments for bone
diseases, a host of new synthetic materials to repair bone fractures or to
serve as bone
replacements, and tissue engineering techniques to induce bone growth from
scratch using a
patient's own bone ceils.

"There is more activity now in this field than there ever has been before,
which I find
gratifying," says Ralph E. Holmes, professor of surgery and head of the
division of plastic
surgery at the University of California, San Diego, Medical Center. Holmes,
who invented a
scanning electron microscope backscatter imaging technique to quantitatively
measure
ingrowth of bone in implants, has been a leader during the past 20 years in
evaluating many


CA 02629555 2008-05-14

synthetic materials. "I think the current activity is a sign of the maturity
of all the sciences
involved and a recognition that not just one thing can make bone heal," Holmes
observes.
Although bone seems lifeless, it actually is made up of a very alive, porous
framework that is
constantly rebuilding itself. Bone is a composite material made up of collagen
protein fibers
threading through hydroxyapatite, Ca5(PO4)30H. Hydroxyapatite makes up about
70% of
bone structure and essentially all of the enamel in teeth. The coliagen
fibers, a bundled array
of cross-linked helical polypeptide strands, provide extra strength, allowing
bones to flex
under stress.

BENEATH BONE'S HARD EXTERIOR...

Bone tissue replaces itself through the action of cells called osteociasts
that produce acids to
dissolve (resorb) hydroxyapatite and enzymes to break down collagen. The
resulting release
of calcium and proteins prompts other cells called osteoblasts to lay down new
matrix that
mineralizes and forms hydroxyapatite and coliagen. Some growth factors, such
as bone
morphogenetic proteins, are manufactured by bone cells themselves to either
increase or
decrease bone remodeling.

Blood vessels snake through the bone framework, too, carrying many different
compounds
that orchestrate bone remodeling, such as calcitrol (a form of vitamin D-3),
calcitonin (a
thyroid hormone that prevents bone resorption), parathyroid hormone (which
works with
calcitrol to regulate caicium and phosphate metabolism), and prostaglandins
(fatty acids that
perform hormonelike functions). Beneath all of that is the bone marrow that
creates the
osteoblasts and osteociasts as well as the red and white blood cells.

Skeletal deficiencies from trauma, tumors and bone diseases, or abnormal
development
frequently require surgical procedures to attempt to restore normal bone
function. Although
most of these treatments are successful, they all have associated problems and
limitations.
For a minor fracture, usually a few weeks in a cast are all that is needed for
the bone to
repair itself. For a more severe fracture or one in a particularly tricky
position, a bone cement
or filler may be used to help strengthen the fractured bone so it heals
faster. These
procedures may or may not require metal hardware such as plates, pins, or
screws for extra
support.

For severe fractures, a bone graft may be needed. In a typical bone graft,
natural bone or a
synthetic material is shaped by the surgeon to fit the affected area and held
in place with
hardware. Over time, the natural bone growth process takes over and at least
partially
resorbs the grafted bone. Key to the level of resorption is the porosity of
the implant material
that allows ingrowth of vascularized tissue.

Surgeons have been performing bone grafts for years. The preferred method is a
procedure
called an autograft, where bone fragments are taken from a patient's own body-
usually a hip
(iliac crest), the pelvis, or ribs-and affixed to healthy bone. An alternative
method, called an
allograft, uses bone donated from a cadaver and works nearly as well.

Bone grafts are painful, complicated procedures that generally involve a long
recovery


CA 02629555 2008-05-14

period. Drawbacks to autografts are the two surgical procedures needed,
meaning longer
hospitalization and recovery time, and higher cost. Besides, the body doesn't
carry much
spare bone.

Allografts eliminate the need for a second surgery, but after sterilization,
the donated bone
loses much of its strength. In addition, when bone grafts are taken from
cadavers, there is
the risk of rejection or of transmitted diseases such as hepatitis B or AIDS
caused by
infection with the human immunodeficiency virus. Allograft procedures have
been on the
decline since late 1993 when the Food & Drug Administration issued regulations
governing
bone and tissue banks because of the risk of transmitting infectious diseases.

A third option, which today is becoming more viable, is use of a synthetic
material to replace
lost bone. Synthetic materials have the advantage of eliminating the need for
surgery to
claim bone for the graft procedure, and eliminating the chance for rejection
or transmission of
infectious disease. An additional benefit is a significant reduction in
medical cost and, in
general, a faster recovery time.

The ideal bone substitute, according to Holmes, would approximate the
autograft, requiring
minimally that it be biocompatible and osteoinductive so that the body's
natural bone-making
process eventually would replace the implanted material. Most synthetic
materials, however,
are weakly resorbed or do not resorb at all. But low resorption is not
necessarily a drawback,
he says.

In some applications, such as a long bone fracture, one might hope for
complete resorption
of a synthetic material. But in other cases, such as reconstruction of the
chin, resorption may
not be desirable at all. Thus, a combination of autograft and synthetic
material with hardware
often turns out to be the best solution, Holmes notes.

The driving force to develop new treatments for bone diseases, the fractures
associated with
them, and fractures from trauma is that they make up an important segment of
the health
care industry. Americans suffer some 5.6 million fractures, and surgeons
perform about 3.1
million orthopedic procedures in the U.S. each year, according to the American
Academy of
Orthopaedic Surgeons. Industry experts expect these numbers to increase
steadily as the
general population grows older.

In 1995, the latest year for which statistics are available, there were some
426,000 bone-
graft procedures in the U.S., according to Medical Data International (MDI), a
market
research firm based in Irvine, Calif. That makes bone second only to blood
transfusions on
the list of transplanted materials.

The worldwide bone-graft market was estimated to be about $800 million
annually, about half
in the U.S., MDI reports. In 1995, about 58% of bone-graft procedures were
autografts,
compared with 34% allografts, and 8% synthetic materials, and nearly half of
the procedures
involved the spine. The percentage of synthetic procedures is expected to rise
substantially
as new biomaterials continue to come onto the market.

Treating bone diseases
When osteociasts dissolve old bone faster than osteoblasts can replace it, the
result is
osteoporosis. Osteoporosis is characterized by loss of bone density that can
lead to
debilitating fractures of the hip and spine. The disease usually strikes
postmenopausal


CA 02629555 2008-05-14

women (age 50 to 70) as a result of decreased estrogen and progestin levels.
The disease
also can occur later in life (after age 75) in women and, as a result of
testosterone
imbalance, in men.

According to the National Osteoporosis Foundation, an estimated 20 million
women in the
U.S. have osteoporosis. That number is projected to grow to more than 35
million by 2015.
Furthermore, the financial impact of osteoporosis in the U.S. has been
estimated to be
almost $14 billion per year in medical costs and lost productivity. These
sobering statistics
are the driving force for research into drugs to prevent or treat osteoporosis
and other bone
diseases. Industry analysts expect the global osteoporosis market for
pharmaceuticals to
reach $5 billion by 2005.

Osteoporosis can be detected by radiological imaging techniques used to
measure loss of
bone density. Besides prevention through diet, a number of therapies are being
used to treat
or prevent the disease. Estrogen replacement therapy, approved in 1988 by FDA,
is
common. Combined estrogen-progestin therapy as well as calcitonin therapy also
are being
used.

Diphosphonates are a class of compounds that are now becoming available in the
U.S. as
nonhormonal drugs for prevention of osteoporosis and as a treatment to help
prevent bone
fractures in patients who already have osteoporosis. The drugs are also
proving successful
for treatment of Paget's disease of bone (a chronic dissolving of normal bone
followed by
disorganized, enlarged, and weakened bone formation) and heterotopic
ossification
(abnormal overgrowth of bone, usually at postoperative sites such as the hip).
Although the
exact mechanism of action of diphosphonates is unclear, they accumulate on
bone surfaces
and inhibit osteoclast resorption, allowing the bone-formation process to
dominate.

DIPHOSPHONATES HELP BOOST BONE DENSITY

The use of diphosphonates to treat bone disease was first advanced by emeritus
research
chemist M. David Francis and coworkers at Procter & Gamble in Cincinnati.
Francis' other
research accomplishments include discovering the benefits of adding fluoride
salts to
toothpaste to prevent tooth decay and adding pyrophosphate (P407 4-) for
tartar control
(C&EN, March 11, 1996, page 34).

Francis led the development of etidronate (disodium 1-hydroxyethane-1,1-
diphosphonate), a
compound that Procter & Gamble originally had looked at as a detergent
additive to chelate
calcium and other hard-water ions. Etidronate, now marketed by Procter &
Gamble as
Didronel, is used in the U.S. and abroad to treat Paget's disease and
heterotopic ossification.
Procter & Gambie also markets Didronel in 17 countries for osteoporosis.

However, in April of this year, Merck announced that it had become the first
company to
receive final U.S. approval for a diphosphonate to help prevent or treat
osteoporosis.
Alendronate (sodium 4-amino-l-hydroxybutylidene-1,1-diphosphonate) is now
marketed in


CA 02629555 2008-05-14
49 countries as Fosamax.

FDA's decision to clear the drug, according to the company, was based
primarily on two-year
results of an ongoing six-year study of more than 1,600 women ages 45 to 59.
The drug is
working as well as hormone replacement therapy to increase bone mass at the
hip and
spine, while a placebo group is showing a gradual loss of bone mass.

Procter & Gamble has another diphosphonate compound, risedronate (sodium 2-(3-
pyridinyl)-1-hydroxyethylidene-1,1-diphosphonate), that is in late-stage Phase
III clinical trials
for treatment of osteoporosis and Paget's disease. In March, the company filed
a new-drug
application with FDA for treatment of Paget's disease, and plans to file
another application for
treatment of osteoporosis in 1998. In May, Procter & Gamble announced it was
forming a
global alliance with Hoechst Marion Roussel-the pharmaceutical company of
Germany's
Hoechst-to commercialize the new drug under the name Actonel.

Mending and making bones
Poly(methyl methacrylate), an old standard, has been used for decades as a
synthetic filler
to repair skeletal defects and affix metal implants to bone. Usually methyl
methacrylate is
polymerized in situ at the site where additional bone is needed. The polymer
hardens to
become stronger than bone and is generally a good substitute.

One of the advantages of poly(methyl methacrylate) as well as similar types of
bone fillers
and cements is that they can be injected through the skin. But there are a
couple of problems
critical to the use of these materials. One is that they usually must cure in
situ and in doing
so can generate heat that could damage surrounding soft tissues. Another, and
perhaps
more important issue, is that most of the materials are minimally degradable
or don't degrade
at all, nor can they support ingrowth of new bone tissue.

The concept of developing materials that are biocompatible and can at least be
partially
resorbed in the natural bone-growth process grew out of the ground-breaking
research in the
1960s by Marshall R. Urist, a professor of orthopedic surgery at the
University of California,
Los Angeles.

Urist was the first researcher to conclusively demonstrate the phenomenon of
osteoinduction, or the natural process of bone desorption and formation. For
more than a
century, surgeons had recognized the ability of demineralized bone to aid bone
healing. But
it was Urist's work on implanting demineralized bone segments into animals and
being able
to induce new bone growth within the implants that led him to conclude in 1965
that cells in
the bone matrix stimulated cells at an implant site to differentiate into
osteoblasts and
osteoclasts. He later showed that bone morphogenetic proteins are the sole
inducers of bone
cell differentiation. His original paper reporting those findings was recently
reprinted as a
landmark paper in the Journal of NIH Research [9, 43 (1997)].

Urist's findings coupled with better diagnostic techniques subsequently gave
rise to a greater
understanding of bone structure and the bone-formation process. This
understanding, in turn,
led to the development of a host of calcium-based synthetic bone products
designed to
mimic natural bone and to actually be resorbed by the body. It has been only
recently, after
many years of animal trials, that these materials have advanced to the point
that they are
acceptable for use in humans and are starting to gain FDA approval.


CA 02629555 2008-05-14

One company well ahead in the synthetic bone implant market is Interpore
International of
Irvine, Calif. In November 1992, Interpore became the first company to receive
FDA approval
for a synthetic bone-void filler.

The company's Pro Osteon hydroxyapatite is made from coral through a
thermochemical
process developed in the 1970s. Currently, it is the only synthetic product on
the market that
has a porous infrastructure similar to natural bone. The interconnected
structure of the coral
remains intact throughout processing, providing a matrix through which blood
vessels and
new bone tissue can grow.

INTERPORE'S SYNTHETIC BONE GRAFT...

Interpore acquires between 2 and 4 tons of coral each year from atolls in the
Pacific and
Indian Oceans to make its product, less than 1% of the total annual amount of
coral imported
into the U.S., the company says. The amount of coral harvested for import is
controlled by
the Convention on International Trade of Endangered Species of Wild Fauna &
Flora
(CITES) and is generally agreed upon by ecologists to present little threat to
fragile reefs as
long as the coral is harvested in ways that sustain the reefs as living
structures. One coral
"head" weighing 150 to 200 lb provides enough material for several hundred
bone grafts.
The synthetic material is prepared by heating the coral-which is essentially
calcium
carbonate-with ammonium phosphate at more than 200 C for 24 to 60 hours to
obtain about
95% hydroxyapatite. The material is processed into block or granular form and
sterilized by
gamma radiation.

Used similarly to natural bone in autograft procedures, the synthetic material
has about the
same length of healing time. A surgeon can shape a block of the material, for
example, to fit
into a fracture crevice or a carved out portion of a long bone. The graft area
is then stabilized
with a metal plate and screws, which later can be removed.

The natural porosity of the material does have the drawback of reducing its
strength, notes
David C. Mercer, Interpore's president and chief executive officer. But the
porous structure
provides room for bone tissue to immediately grow into the pores of the
implant. However,
the material is only partially resorbed and replaced by natural bone. The
company is now
evaluating in preclinical studies a related new product that has a higher
resorption rate.
Pro Osteon is currently approved for nonweight-bearing treatment of fractures
at the wide
end of long bones and for jaw and reconstructive facial surgery, according to
Mercer.
However, the material has been used in many cases to replace a short section
of a long
bone, he says. In such cases, the limb must be immobilized for a long period-
perhaps
several years-to ensure new bone growth has become strong enough to support
weight.
Sales of Pro Osteon have increased steadily since 1992, reaching $11.7 million
in 1996, up
47% from the previous year. Sales in the first half of 1997 continue to be
strong, increasing
13% from last year's first half to $6.3 million. lnterpore also has approval
to sell its bone
substitute in 41 countries and began international marketing in 1995.


CA 02629555 2008-05-14

Although synthetic biomaterials on the market or under development work well
for their
intended functions, none has yet proven to be sufficiently strong or able to
be processed into
a large enough piece to stand in as a complete replacement for long bones.

Research by University of Texas, Austin, chemistry professor Richard J. Lagow,
however,
appears to have come the closest to that goal. Lagow makes a high-purity
hydroxyapatite
from scratch by reacting calcium metal, calcium hydroxide, and phosphoric acid
at 700 to
850 C. Lagow also has come up with analogous high molecular weight linear
calcium
polyphosphates by reacting hydroxyapatite with phosphoric acid. The success of
his
research has led to the creation of a small Austin-based company, called
OsteoMedica Inc.,
to develop the synthetic materials as potential complete bone substitutes.

Key to developing these compounds for implant materials, Lagow says, is their
high purity,
which does not retard bone growth. Also important, he says, is holding the
reaction
temperature below the 1,200 C temperature at which calcium phosphate fuses
into a
ceramic.

Besides controlling bulk size and shape, during his proprietary synthesis and
processing
method, Lagow can moderate the interconnecting porosity of the synthetic
material-in the
range of 150- to 400-pm pores-to match the density of different types of bone.
"Interconnecting means that the body can vascularize it quickly and bone can
then grow into
the material much faster because there is a greater surface area for the
osteoblasts and
osteoclasts to work," Lagow notes. "Otherwise, the osteociasts must tunnel
through the bone
matrix to resorb the synthetic material, which is a much slower process."

Lagow has collaborated with UT Austin chemical engineering professor Joel W.
Barlow and
others to develop a selective laser sintering technology that they have
patented to fabricate
complex bone shapes from the calcium phosphate materials.

Barlow and his research group developed a technique whereby they use a spray
drier to coat
hydroxyapatite or calcium phosphate powders with a poly(methyl methacrylate)
that acts as a
binder. The materials readily fuse in the sintering process but maintain their
interconnecting
pore structure. And, unlike other bioceramics being investigated as bone
substitutes, they
retain their high strength during processing, Lagow says.

LAGOW S METHOD TO SYNTHESIZE...

The laser can be guided to form intricate bone shapes by computer, using data
sources such
as magnetic resonance imaging or computed tomography. The molded ceramic is
then
heated to remove the polymer and further processed. Thus far, the researchers
have been
able to generate a wide range of bone sizes and shapes. The technology has
since been
licensed to BioMedical Enterprises Inc., San Antonio, which is pursuing
biocompatability
studies of fabricated calcium phosphate implants.

OsteoMedica's goal is eventually to provide surgeons with synthetic molded
bone "blanks"
that can be custom shaped to fit a patient's needs. The company's bioceramic,
called
Megag raft 1000, so far has been successful in tests replacing tibia sections
in rabbits and


CA 02629555 2008-05-14
other animals.

And in a very successful study on dogs, sections of the radius were replaced
with the
synthetic bone. After 11 months, support plates and screws were removed, and
the dogs
eventually regained full use of their legs. One of the most promising aspects
of the study was
that, some three years after the time of the implant, the synthetic material
was completely
resorbed- results that haven't been reported for other synthetic materials.
OsteoMedica is
preparing to begin clinical trials in humans for spinal fusions in the U.K.
and Australia, where
regulatory requirements aren't as stringent as in the U.S.

"Being able to synthesize novel hydroxyapatite material to make it accessible
to higher bone
ingrowth and remodeling rate is a critical step," says UC San Diego's Holmes,
who has
evaluated both Interpore's and OsteoMedica's products. While he finds both
materials work
well for their intended use, a product such as OsteoMedica's hydroxyapatite
being available
in different pore sizes would be particularly useful for a wide range of
applications to control
the level of resorption, he notes. "Wi'th a range of porosities and resorption
rates, surgeons
could learn to choose the synthetic material for a particular application much
in the way they
choose a suture material."

Glen O'Sullivan, an assistant professor of orthopedics at Stanford University
Medical Center,
also has used lnterpore's product in his patients and has worked on a clinical
study of spinal
fusions in sheep using OsteoMedica's material. He, too, finds both materials
perform well for
their designed use.

"One advantage of OsteoMedica's product is it is one of the hardest
materials," O'Sullivan
notes. "For example, lnterpore's material can be crumbled between the fingers,
while
OsteoMedica's is strong enough to drill holes in it. This makes OsteoMedica's
a good
candidate for use in the spinal column, whereas Interpore's would not be-and
it isn't
approved by FDA for that."

Holmes and O'Sullivan agree that there isn't one material that is going to be
suitable for all
applications. "With any of these new products," O'Sullivan says, "one likes to
be optimistic,
but the hopes and expectations may not always pan out for all applications."
It makes sense,
he adds, that FDA only approves what might appear to be broadly applicable
bioceramic
materials for a narrow range of applications.

There are perhaps dozens of calcium-based synthetic materials in addition to
Interpore's and
OsteoMedica's that have received approval for use in the past couple of years
or are
anticipating approval soon. In 1996, for example, FDA approved OsteoSet, a
calcium sulfate
(plaster of paris) bone-void filler developed by Wright Medical Technology,
Arlington, Tenn.,
that is reasonably resorbed by the body in as little as eight weeks. Another
product cleared in
1996 by FDA for repair of cranial defects is a hydroxyapatite bone cement
developed by
American Dental Association Health Foundation researchers called BoneSource.
The
material will be manufactured by OsteoGenics Inc. and distributed by Howmedica
Leibinger,
a division of Pfizer.

An injectable bone cement developed by Norian Corp., Cupertino, Calif., is
described by the
company as a biocompatible, moldable compound made by mixing calcium
phosphate,
tricalcium phosphate, and calcium carbonate with sodium phosphate solution
into a
toothpastelike substance. Norian currently has approval to market its Skeletal
Repair System


CA 02629555 2008-05-14

in Europe and Canada, and is working toward regulatory approval in Japan and
the U.S.,
mainly for treatment of wrist and hip fractures.

New directions
In 1993, FDA approved one of the first bone graft substitutes that capitalized
on a new
concept for products to facilitate bone repair. Collagraft, marketed by
Bristol-Myers Squibb
subsidiary Zimmer Inc., Warsaw, Ind., is a hydroxyapatite/tricalcium phosphate
and bovine
collagen that must be mixed with a patient's own bone marrow.

In late 1996, Interpore signed a license and development agreement with
Quantic
Biomedical, San Rafael, Calif., for a technology to use a gellike material
containing bone
growth factors from a patient's own blood that can be combined with its coral-
based Pro
Osteon to provide accelerated bone growth. Preclinical feasibility trials are
expected to begin
by the end of this year.

These examples illustrate the impact that the "delayed discovery" of bone
morphogenetic
proteins (BMPs) has had in opening the research community to a new direction:
the concept
that BMPs, bone cells, and various hormones could form the basis of an
engineered system
for bone repair that includes bioceramics or biopolymers. In essence, the
world of
bioceramics is being wed to the world of tissue engineering.

Although Urist's work on osteoinductivity was definitive, most researchers
weren't convinced
until BMPs actually began to be cloned by recombinant DNA methods some 20
years after
his landmark research paper.

In 1988, the first group to clone a BMP was that of senior director John M.
Wozney at
biotechnology company Genetics Institute, Andover, Mass. [Science, 242, 1528
(1988)].
Today, one of Genetics Institute's molecules, BMP2, is in clinical trials for
fracture repair,
spinal fusion, and other possible applications. Several other biotechnology
and
pharmaceutical companies are testing the more than 30 BMPs cloned thus far for
potential
use in bone- and tooth-mending applications. "BMPs are destined to bring
osteogenesis
under the control of surgeons before the turn of the century," Urist noted in
a commentary on
his landmark paper.

"Despite the great advances in the synthetic materials, one still needs the
bone cells,"
O'Sullivan states. "Bone cells are needed in the implant material and you want
a means of
stimulating bone cell activity. This is where research with BMPs is really
going to take off."
O'Sullivan notes that even during autograft procedures, surgeons attempt to
aspirate bone
celis from adjacent bone to incorporate into the implanted bone, and sometimes
use material
extracted from presurgical blood donation by the patient to help induce
implant bone growth
and resorption.

O'Sullivan points out that a critical step remaining for BMPs is to find the
optimum carrier for
implantation. Although bioceramics likely will work well, he says, they do
have the problem of
slow resorption. Thus many researchers believe that biodegradable polymers
will work best
as delivery devices for human growth factors, he says.

One of the leaders in the biodegradable polymer area is Antonios G. Mikos, an
associate
professor in the department of chemical engineering and the Institute of
Biosciences &
Bioengineering at Rice University, Houston. "The advantage of using polymers
is that one


CA 02629555 2008-05-14

can very accurately engineer their mechanical properties and degradation
characteristics,"
he explains. The size and shape of the scaffold can be made to order as well,
depending on
which bone a potential patient may need.

Mikos and his research group are working on strategies to naturally grow bone
from scratch
either in vitro or in vivo by seeding natural or synthetic polymer scaffolds
with bone cells or to
use the scaffolds as conduits to induce new bone growth from surrounding
tissues. Several
substrate materials are being investigated by a number of researchers, Mikos
notes,
including poly(a-hydroxy esters), polyanhydrides, polyimides,
polyphosphazenes, and
collagen.

The success of such strategies is dependent on the scaffold material's being
biocompatible,
osteoconductive, and quickly degradable into products that can be metabolized
or excreted,
he explains. For example, poly(lactic-co-glycolic acid) breaks down to lactic
acid and glycolic
acid, which are metabolized in the body and excreted as carbon dioxide and
water.

Osteoblast transplantation onto a polymer scaffold would eliminate the problem
of donor
scarcity, immune rejection, and pathogen transfer by taking the needed cells
from a patient's
own body, Mikos points out. Although osteoblasts may be obtained by a variety
of methods,
including bone chips from an injury site or enzymatic digestion of harvested
bone, the most
desirable method would be to obtain the cells from the patient's own bone
marrow.
Osteoblasts obtained from bone marrow, for example, can also be expanded in
tissue culture
in a lab and seeded onto a polymer scaffold for implantation.

Poly(lactic-co-glycolic acid) has been extensively investigated as a material
for tissue-
engineering scaffolds because it already has been approved by FDA for use in
surgical
sutures, can be made with controlled pore size, and degrades well. The first
such scaffolds
were designed by biomedical and chemical engineering professor Robert S.
Langer at
Massachusetts Institute of Technology and Joseph P. Vacanti of Harvard Medical
School in
the late 1980s to create an in vitro environment that enables cells to
organize themselves to
form functioning tissues. Langer and Vacanti prepared crude scaffolds by
bonding together
poly(lactic-co-glycolic acid) fibers into a two-dimensional network.

In 1991, working with Langer and Vacanti, Mikos (then at MIT) further
developed the polymer
scaffolds by incorporating sodium chloride crystals into the copolymer matrix
by adding
crystals to a solution of the dissolved polymer. The salt crystals were later
leached out,
leaving behind a porous polymer matrix. Mikos was able to control porosity and
pore size by
varying the concentration and size of the crystals.

In another technique, Mikos extruded polymer fibers and aligned them in the
shape of the
desired scaffold. He embedded the arrangement in a polymer with a higher
melting point and
bound the scaffold together by heating. After cooling, he selectively
dissolved the embedding
medium, leaving behind an interconnected, highly porous structure.

Mikos, graduate student Susan L. Ishaug-Riley, and coworkers have recently
conducted
feasibility studies to show that bone formation in vitro and in vivo is
possible by culturing rat
osteoblasts in three-dimensional poly(lactic-co-glycolic acid) foams of
different pore sizes
(shown on the cover of this issue).

In one study, the polymer foams supported the proliferation of the seeded rat
osteoblasts in
--i


CA 02629555 2008-05-14

vitro to form a calcified bonelike tissue after two months [J. Biomed. Mater.
Res., 36, (1997)].
The goal of the study was to gain a better understanding of the important
parameters in the
design of an osteoblast foam-culture system before attempting osteoblast
transplantation in
vivo.

In a subsequent in vivo study, rat bone marrow osteoblasts were seeded onto
polymer foams
and implanted into the rat mesentery (the membrane of the abdominal cavity)
[J. Biomed.
Mater. Res., 36, 1 (1997)]. Growth of islands of mineralized bonelike tissue
in the foam
surrounded by fibrovascular tissue was observed within one week and had
significant
penetration of bone tissue into the scaffold after seven weeks.

The findings were encouraging, the researchers note, because they indicate
that the
regenerative potential of the seeded polymer scaffolds for new bone growth
with transplanted
cells and secreted bone growth factors may further induce bone growth from
adjacent bone.
Ishaug-Riley received a student outstanding research award from the Society
for
Biomaterials for the in vivo study at the society's 23rd annual meeting in New
Orleans in
May.

Osteoblast transplantation is not a straightforward approach, Mikos says,
noting that
because bone is highly vascularized, it is not possible to engineer and grow a
complete bone
or bone fragment in vitro and transplant it. "The maximum thickness of new
bone one can
create in vitro is a few hundred micrometers, which is not significant for
clinical applications.
However, the goal is to form new bone tissue in vivo and not in vitro. Then,
vascularization
becomes equally important to bone formation and necessary for regeneration."

A critical issue for cell transplantation is which phenotype of transplanted
cell should be
used. "It is not clear if one should transplant osteoblasts or preosteoblasts
or progenitor
cells," he says.

Mikos believes it will be possible in the next decade for tissue-engineered
implants to be
used for the reconstruction of skeletal deformities resulting from trauma,
tumors, or abnormal
development. "I hope that new cell-based therapies will be developed for the
treatment of
osteoarthritis and osteoporosis based on combinations of degradable
biomaterials, growth
factors, and cells," he says. "Yet, the main drawback with new polymers is the
time and effort
needed to get FDA approval for their use."

O'Sullivan is optimistic about the prospects of tissue engineering in bone
repair, but also
cautious about a couple of potentially critical problems. FDA currently is not
certain about
how to regulate tissue-engineered products, he says. (A problem with tissue
cultures is
potential contamination with a fungus, bacteria, or mold.) "And it will be
interesting to see if
the new technology will become available in an affordable manner, given that
the couple of
companies working on BMPs have spent a tremendous amount of money during the
past few
years to develop a research infrastructure." He thinks the BMP companies will
end up
controiling the technology development path for the synthetic implant
companies.

"The next step is going to be a fine balance between cost and whether the
outcome is going
to be worth it."

n coral that is converted to hydroxyapatite and processed into block or
granular form. X-ray ima
)one fragment was removed and replaced by the synthetic material. Metal plates
and screws ob


CA 02629555 2008-05-14
-rt the ankle until it heals.

hemCenter ^Pubs Div. Home Page

We are so looking at cloning fish (especially high in omega 3) and the cells
that produce or
are rich in omega 3 for food any and all varieties, any and all climates for
sushi, chinese
seafood cuisine, fish and chips and any and all cuisine, including
pelletisizing for food for
livestock, also cloning the silivary glands that produce bird's nest soup, and
jellyfish dishes,
as well as clone any and all livestock (including chicken GP 33%), for flesh
as well as fur and
for leather and skin...

(3.5%)
Growing com on the top layer could package (hereinafter, package/packed refers
to
biodegreadable absorbant yet stiff enough to support planst such as corn)
large tight rooting
and the stalk stands up - packed surrounding the roots is enough soil and/or
peat and/or
manure/compost/partially treated sewage and any and all organic wastes and/or
chitin/chitosan to provide for the corn (and/or any and all plants to last
through maturation -
the packages fit in holes in large styrafoam trays and/or lava rock, and a
further layer of lava
rock below. Underneath the lava rock is a perforated strong waterproof
material (eg. PVC) to
hold up the two layers above and finally the bottom is a catch basin, where we
could grow
lobsters/crawdads/crayfish/tilapia/any and all shell fish/and any and all
fishes and any and all
water creatures; the entire system could be flushed with UV treated salt water
and re-
circulated UV treated salt water and low concentration feed tea
manure...Alternatively if the
tea manure has too much nitrogen, we could skip the middle layer of lava rock
and rather
plant the com in tight packages with enough peat and top soil in the rooting
package
surrounded by either a thick layer of styrafoam with holes that fit the
rooting packages, and
or lava rock to keep the corn from tipping and then place a membrane -
multiple layer (eg.
sealing above the perforated strong waterproof material eg. PVC) to keep the
nitrogen out,
from entering the catch basin where the lobsters/crawdads/crayfish/tilapia/any
and all shell
fish/and any and all fishes are housed or more importantly to keep the salt
water below from
tainting the corps above, unless the farmed creatures below are fresh water
species.
Alternatively, we could design the system where water does not pass freely
between the
crops hydroponics' in packages surrounding the stalks of eg. corn we could put
bags of algae
(that are passed with tea manure) above and the farmed water boume species
below we
could recycle water, where nitrogen-ammonia water caused by feces/uneaten are
treated
with UV and then micro organisms in sand and membrane to take out the salt
which
produces fresh water that is re-circulated to the plants/crops above, then the
opposite the
water from the crops (that have nitrogen removed by the roots of the crops)
can be mixed in
salt if for salt water re-circulation. This rotational re-circulation, may
create symbiotic
synergies. We could grow these stacked farming systems on further floors up
all the way up
tall buildings... possibly with TALL GRAVITY TO ELECTRICITY INVENTION buried
underground to provide energy. We could also grow root foods in third world
countries (as


CA 02629555 2008-05-14

well as rice) such as yam, sweet potatoes and potatoes which may be higher
yield than rice
- in its ability to fill stomachs (even gingseng)... by packing the area
surrounding the roots
with a large footprint of tightly peat/top soiVmanure/slightly treated
sewage/compost/and and
all organic waste, perhaps surrounded by a burlap (or biodegradable - any and
all covers) to
wrap, and placed in a hydroponic solution (eg. tea manure; possibly supplement
with
phosphorous/potassium and vitamins), surrounded by support such as thick
styrafoam flats
to support the stock and lava rock surrounding the packaged roots - with ample
packaging of
root growth space so as not to impede the growth of root crops - possibly in
sky scrapper
green houses.

We could also methylation, imprinting to increase harvests of any and all
crops including any
and all bio crops.

We could coat any and all seeds (especially bio crops seeds) as well as pack
chitin/chitosan
around roots.

In all of the above we use chitin and chitosan interchangeably.

The some of the following can be interchangeably between any and all creatures
and any
and all plants.

We are culturing stem cells (using testing for the best timing and
sequences...) and
enhancing their proliferation by using GM-CSF (granulocyte-macrophage colony-
stimulating
factor) + fetal bovine serum (FBS) (and/or any other mediums) to methylate in
the p15 CpG
island. Viable cells were responsible for this epigenetic change. Following
the GM-
CSF + FBS application the culture was added to inhibitors for DNA
methyltransferase
(DNMT) and histone deacetylase (HDAC) caused the demethylation of nearly all
CpG sites in
the p15 CpG island on every allele sequenced. GM-CSF may be able to induce de
novo
methylation of the p15 gene, using HDAC(s) as well as DNMT(s).

Taken Wikipedia:

Neoplastic tumors often contain more than one type of cell, but their
initiation and continued
growth is usually dependent on a single population of neoplastic cells. These
cells are
usually presumed to be clonal - that is, they are descended from a single
progenitor cell.
The neoplastic cells typically bear common eg netic or epigenetic
abnormalities, an evidence
of clonality. For some types of neoplasm, e.g. lymphoma and leukemia, the
demonstration of
clonality is now considered to be necessary (though not sufficient) to define
a cellular
proliferation as neoplastic.

Nocodazole and Colchicine have the opposite effect, blocking the
polymerization of tubulin
into microtubules, we could try these drugs to increase the proliferation of
stem cells.

We are also using proto-oncogenes to increase stem cell proliferation (see
below taken from
Wikepedia).

Proto-oncogene


CA 02629555 2008-05-14

A proto-oncogene is a normal gene that can become an oncogene due to mutations
or
increased expression. Proto-oncogenes code for proteins that help to regulate
cell growth
and differentiation. Proto-oncogenes are often involved in signal transduction
and execution
of mitogenic signals, usually through their protein products. Upon activafion,
a proto-
oncogene (or its product) becomes a tumor inducing agent, an oncogene.u
Examples of
proto-oncogenes include RAS, WNT, MYC, ERK and TRK.

edit Activation

The proto-oncogene can become an oncogene by a relatively small modification
of its
original function. There are three basic activation types:

= A mutation within a proto-oncogene can cause a change in the protein
structure,
causing
o an increase in protein (enzyme) activity
o a loss of regulation
= An increase in protein concentration, caused by
o an increase of protein expression (through misregulation)
o an increase of protein stability, prolonging its existence and thus its
activity in
the cell
o a gene duplication (one type of chromosome abnormality), resulting in an
increased amount of protein in the cell
= A chromosomal translocation (another type of chromosome abnormaiity),
causing
o an increased gene expression in the wrong cell type or at wrong times
o the expression of a constitutively active hybrid protein. This type of
aberration
in a dividing stem cell in the bone marrow leads to adult leukemia

Mutations in microRNAs can lead to activation of oncogenes.v New research
indicates that
small RNAs 21-25 nucleotides in length called microRNAs (miRNAs) can control
expression
of these genes by downregulating them.r7l

edit Proto-Oncogenes

There are several systems for classifying oncogenes, 8 9 but there is not yet
a widely
accepted standard. They are sometimes grouped both spatially (moving from
outside the cell
inwards) and chronologically (paralielling the "normal" process of signal
transduction). There
are several categories that are commonly used:

Category Examples Description

Usually secreted by specialized
cells to induce cell proliferation in
themselves, nearby cells, or distant
Growth factors, or c-Sis cells. An oncogene may cause a cell
mitogens to secrete growth factors even
though it does not normally do so. It
will thereby induce its own
uncontrolled proliferation (autocrine
IOog), and proliferation of


CA 02629555 2008-05-14

neighboring cells. It may also cause
production of growth hormones in
other parts of the body.

Kinases add phosphate groups to
other proteins to turn them on or off.
Receptor kinases add phosphate
groups to receptor proteins at the
epidermal growth factor receptor surface of the cell (which receive
(EGFR), platelet-derived growth protein signals from outside the cell
Receptor tyrosine factor receptor (PDGFR), and and transmit them to the inside
of
kinases vascular endothelial growth factor the cell). Tyrosine kinases add
receptor (VEGFR), HER2/neu phosphate groups to the amino acid
tyrosine in the target protein. They
can cause cancer by turning the
receptor permanently on
(constitutively), even without signals
from outside the cell.

Src-family, Syk-ZAP-70 family, and
Cytoplasmic BTK family of tyrosine kinases, the _
tyrosine kinases Abl gene in CML -
Philadelphia chromosome
Cytoplasmic
Serine/threonine Raf kinase, and cyclin-dependent kinases and their kinases
(through overexpression).
regulatory subunits

Regulatory Ras protein
GTPases -
Transcription
-
MYC- gene
factors
We plan to add
Cargo transport

In the cell, small molecules such as gases and glucose diffuse to where they
are needed.
Large molecules synthesised in the cell body, intracellular components such as
vesicles, and
organelles such as mitochondria are too large (and the c osol too crowded) to
diffuse to
their destinations. Motor proteins fulfill the role of transporting large
cargo about the cell to