Aplastic Anemia Research Center

This page includes my research prior to 2005.

Click here to access Active Research Blog.

Overview of This Page

I am convinced that there is a way to cure Aplastic Anemia and the answer lies in understanding why and how the immune system decides to attack immature cells.  Finding a cure will require the understanding of the various disciplines listed below and I will use this area of the site to store everything I am currently researching.  

I will constantly try to put everything into layman's terms.  My mission is to simplify, simplify, simplify!  People who constantly try to mystify this stuff are exactly like the computer professionals who try to keep people in the dark - the old mushroom theory (keep 'em in the dark and feed 'em shit).  Well, I refused to be kept in the dark in the world of data processing and I now refuse to be kept in the dark relative to my disease!

As a starting point, my initial bone marrow aspirate and smears revealed "mildly hypercellular marrow with mild hyperplasia or erythrocytic series and hypoplasia of megakaryocytes."  This means that my marrow has a slightly larger number of cells in a cross section of the marrow than is normal and an abnormal multiplication or increase in the number of normal cells with a lower number of megakaryocytes. Megakaryocytes are the giant polyploid  cells (the cells containing the genetic structure or chromosomes necessary for producing additional cells) of bone marrow that gives rise to 3-4,000 platelets each.  So in essence, I have a reduced number of the cells necessary to create platelets. 

If anyone reading this sees information that is incorrect or misinterpreted, please send me a note of correction. Thanks. 

BTW, much of my cursory research while I am traveling will now be at the forum.  It's easier to just send my comments there rather than opening and editing the website.

The Disciplines Involved

In addition to the obvious (clinical trials, treatment protocols and survivability), I am trying to understand the following disciplines as they relate to AA:

• Biology
• Immunology  Another Good Site for Immunology
• Immunopathology
• Biochemistry
• Hematology
• Microbiology
• Genetics and DNA Research
• Human Genome Project
• Multiple Chemical Sensitivities
• Alternative Medicines
• Nutrition and Allergies
• Stem Cells
• Pathophysiology

Other links on the links page. Which I will probably categorize at some point.

http://www.health.sa.gov.au/CanCare/REFERENC/Blood/Hemo/NetIt4/22020x.htm

How to Start - The Scientific Method

Above link is a good place to start by understanding how blood cells are produced.  Dr. Rea said that the biopsy from Strong said I had no Megakaryocytes (A megakaryocyte is the "progenitor" of platelets - one megakaryocyte produces 4000 platelets) present - That is a serious concern as Megakaryocytes is what produces platelets.  No Megakaryocytes, no platelet production!  I asked Sharon to send the biopsy results from Strong to EHCD.  

Nobody mentioned megakaryocytes to me until Dr. Rea came along.  Now I understand that my problem is more at the level of megakaryocytes than it is at the platelet level.  If I could keep my body from destroying megakaryocytes, it would then produce platelets???  Now that I think about it, Dr. Kirsher talked about "immature platelets" and he must have been referring to megakaryocytes (thank god somebody speaks english).

So now when I read Dr. Young's pathophysiology, it is beginning to make a little more sense.  As Richard Laughlin told me a long time ago, the key to understanding what you read is to not let a single word go by that you do not understand.  I literally have to look up every word in his report and then look up the words used to define what I looked up before the fog lifts.

I believe that each of these disciplines plays some part in developing a thorough understanding of the autoimmune and blood related illnesses.  It is a huge undertaking and I am being bombarded with new terminology in each discipline but I will be attempting to sort it out as I go and this is where it will start.  If you know anything about these areas and can help me sort it all out, please send me a note mailto:sales@aebiz.com.

Genetics Glossary - http://www.nhgri.nih.gov/DIR/VIP/Glossary/pub_glossary.cgi

1. Observe the situation
2. Ask a question
3. Turn that question into a testable hypothesis
4. Predict the outcome of your experiment
5. Perform your experiment
6. Analyze the results
7. Evaluate your hypothesis

Hematology Terms - Some Fundamentals Before we Ask the Questions

A Hematology Glossary

Red Cells, also known as erythrocytes, carry oxygen from the lungs to the tissues.

White Cells, also known as leukocytes, are responsible for killing any microorganisms that invade the body.

Platelets are small cellular particles produced in the bone marrow by shedding from very large cells called megakaryocytes and serve as the first line of prevention of bleeding when a blood vessel is damaged.

Anemia - Too few red blood cells in the bloodstream, resulting in insufficient oxygen to tissues and organs

Pancytopenia  Deficiency of all cell elements of the blood, aplastic anaemia.

hypoplastic anaemia  A low red blood cell count that results from the underproduction of red blood cells by the bone marrow. This is often secondary to a drug (chemotherapy) side effect.

thrombocytopenia  A decrease in the number of platelets in the blood, resulting in the potential for increased bleeding and decreased ability for clotting.

granulocytopenia  A reduced number of white blood cells in the circulation.

leukopenia Abnormal decrease in the number of white blood cells.

Polycythemia is the term used to designate overproduction of red cells, white cells and platelets.

Erythrocytosis is the term used to designate the overproduction of red cells alone.

Leukocytosis is the term used to designate overproduction of white cells alone.

Thrombocytosis (or thrombocythemia) is the term used to designate overproduction of platelets alone

Myelofibrosis is the term used to designate an increase in the fibrous tissue of the bone marrow. Myelofibrosis is not a primary process but is always caused by another disorder.

Hematopoietic Progenitor Cells are the parent cells in the bone marrow for red cells, white cells and megakaryocytes. The most primitive hematopoietic progenitor cells are multipotent and give rise to the progenitor cells for red cells, white cells and megakaryocytes.

Myeloproliferative Disease is the term used to define a disease arising in a hematopoietic progenitor cell that results in the uncontrolled (autonomous) overproduction of normal-appearing blood cells in the absence of an appropriate stimulus such as lack of oxygen for red cells, lack of microbial invasion or inflammation for white cells and lack of bleeding for platelets.

Idiopathic (or Essential) is a term used to indicate that the cause for a disease process or disorder is unknown

Clonal is a term used to describe diseases arising from a single cell.

Cytogenetics is a technique used to analyze the number and integrity of a cell's chromosomes

Bone Marrow Aspirate is a technique, similar to drawing blood, for obtaining bone marrow for microscopic examination, cytogenetics and flow cytometry.

Bone Marrow Biopsy is a technique by which a piece of bone containing marrow is obtained when marrow cannot be aspirated to identify the presence of myelofibrosis, and to assess marrow cellularity and architecture.

Flow Cytometry is a technique by which individual blood or marrow cells can be analyzed for clonality.

Uric Acid is a by-product of DNA that can accumulate and cause kidney stones or gouty arthritis if the white count is high or if white cells are being rapidly destroyed by chemotherapy.

Phlebotomy is the removal of blood from a vein to reduce the number of red cells and induce iron deficiency to slow their reaccumulation.

The Questions

I am beginning to believe that what will be most important in my battle is clearly asking the right questions and then researching the answers, so here are some questions I am working:

In acquired aplastic anemia, clinical and laboratory observations suggest that this is an autoimmune disease.  http://aplasticcentral.com/aplastic_facts.htm

Ongoing reading but some fundamentals - At a cellular level, the immune system is charged with remembering microorganisms that are causing problems and develops antigens to destroy these rogue microorganisms.  If the blood cells become targets of the immune system (or apoptosis which is programmed death of unnecessary cells), then the blood cells are in trouble because the immune system is very good at what it does - protect the body from microorganisms it doesn't like (It even remembers that you had chicken pox as a kid and if any small pox like microorganisms show up it destroys them) 

3. What exactly is happening when the lymphocytes attack the immature platelets and how can I follow that thread to understand the immune system malfunctioning - what is Dr. Rea doing to stimulate platelet growth? 

Lymphocytes are responsible for fighting infections and it appears that in Aplastic Anemia, our lymphocytes have decided that the progenitor cells are an infection?  There must be some connection between the fact that I am low in protein and CD34 is a "glycoprotein.

4. Better yet - make sure I understand in detail lymphocytes, CD34 and T cells and apoptosis, etc. and then formulate the questions.

Thanks to Kenton, gaining a much better understanding, but still have a ways to go.

http://www.bioscience.org/news/scientis/apoptos.htm has a good discussion of apoptosis vs. tumor necrosis factor (TNF) apoptosis involves a single cell. TNF involves a group of cells. 

5.How does environmental illness and MCS relate to Aplastic Anemia?

Best guess so far (9/30/01) is that in addition to inheriting a pre-dispositon to an autoimmune disease, my body's immune system began to malfunction as a result of being continually exposed to a series of viruses (shingles, menangitis) and toxins (pesticides, mercury, paint, fertilizer, varnishes, molds).  If we can isolate the causes by testing what my body may react to, we can  prepare and inject antigens to correct the imbalance.  At the same time, I do the physical therapy (exercise, sauna, vitamins and massage) to rid my body of toxins -so far, it appears to be working.  I am also eating a restrictive diet and taking a large number of vitamins and supplements designed to reset my body's immune system.  

6. So what are these CD34 things anyway and how do they interact with everything else we know?

See Kenton's Research.

7. What is the difference between leukocytes and lymphocytes?

A leukocyte is any kind of White Blood Cell.  A lymphocyte is a specific type of WBC in the lymphatic system (The tissues and organs (including the bone marrow, spleen, thymus and lymph nodes) that produce and store cells that fight infection and the network of vessels that carry lymph.)  So - lymphocytes are part of the immune system that fight infections. (how can they keep saying that hodgkins and AA are unrelated??????)

8. I don't understand the fact that if my immune system is over-reacting, why I want to "strengthen" my immune system. As Sue explains it, my immune system is out of balance and the EI strategy is designed to get it back in balance.  Need to further articulate this in a way that I understand.

According to Docs (Lancet and Rea) what I need to do is "balance" my immune system and so far I think the MCS/environmental approach appears to have a better/less toxic way of doing this.

9. Define, rationalize, categorize, synthesize and understand cytoxins, cytokines, proteins, amino acids, CD's, folic acid, interleukins, lymphocytes, leukocytes, toxins, antibodies, antigens, magnesium, calcium, hemoglobin, hematocrits, phagocytosis, neutropenia, thrombocytopenia, T Cells, B Cells, Killer T Cells, apotosis, fibromyalgia, immune system, thymus, tonsils, adenoids, spleen, allergies as related to immune disorders, CFS (Chronic Fatigue Syndrome, MCS (Multiple Chemical Sensitivities), detoxification vs chelation, depuration, mitosis, vitamins (B12, E, C, A), L-Lysine, Cod Liver Oil, seratonin, glycoprotein, lymphatic system vs lymphocytes vs lymph nodes?

Cytokines are chemical messengers that activate the immune system.

10. Why do all these other diseases keep popping up and what is the relationship with AA: AIDS, HIV, Arthritis, Lupus, Hodgkins, TTP, ITP, MDS-RA, CFS, MCS, Diabetes.  

They are all related to the immune system.  If you start by getting the immune system in  balance (see above) then you can begin to address the specific issue that got your immune system out of whack in the first place.  I question why everyone (and especially NIH) fails to recognize the similarities and have a centralized autoimmune effort going on rather than all the fractionalized efforts.  It sucks that HIV gets all the attention when you consider how most people become infected in the first place and by the way I live in fear of one of my almost 100 transfusions passing HIV or hepatitis or some other ugly disease on to me.

Specific difference between AA and ITP courtesy of Kenton:

ITP involves the direst attack on the platelets.  In AA the low
platelet counts are due to the lack of production (maybe in
combination with direct attack) of the platelets.  I a platlet
transfusion was given to someone w/ ITP the platelets would be
marked with antibodies and subsequently destroyed.  So
transfusions are not effective at combatting ITP.  In AA it seems
that the precursor cells to platelts (and other blood cells) are
attacked and decreased in number.  This causes less of these
cells to be present in the body because there is nothing left to
make them.  It seems thaat splenectomy is a common and effective
(though somewhat radical) treatment for ITP.  This gets rid of
the problem (the spleen is involved in the removal of blood cells
from the body) but still does not give us a cause, Why is the
spleen misbehaving?  According to the article that you sent me
there are numerous causes with a similar result (ITP).  In my
mind these should all be considered different diseases, this
would simplify the diagnosis and perhaps aid in the treatment.
The disease could be grouped by common cause and the effective
treatment for each cause could be systematically tried.  There is
no use trying to combat a genetically induced form of ITP with a
treatment that is more consistantly effective at combatting viral
or environmentally induced ITP.  Keep in mind that the body is
incredibly complex at the biochemical and molecular level and the
substitution or deletion of just one enzyme in different
metabolic processes might lead to the same result even though the
difference isn't even related.  The same can be and is true for
proteins or sugars (riboses) in DNA at the molecular level.  One
small difference can make a huge impact on the function of a
gene.  All because two people have the same symptoms it doesn't
mean that they have a disease originating from the same cause.  I
hope that this answers your question about the difference between
AA and ITP.  I'm sending the web site from NIH, you've probably
already been there but it seems generic enough to understand.
Another interesting note about ITP is the reference to anti-D
immunoglobin.  This is another antibody (biologic) that is
involved in Rh recognition and desensitization.  The
effectiveness of this treatment makes me point toward differences
in Rh complexes present on the platelet cells.  Kind of like I
thought CD34 in your AA, as compared with others AA that might be
affected by other receptors.  The fact that the platelets are
marked by the antibodies that result in the removal of the cells
indicate to me that the spleen is not malfunctioning and that the
cells that are marking the cells for destruction are the problem
(usually B-cells) but I do not know the mechanism that the spleen
uses for removal of the cells.  Something more for me to look up,
in any cae I would have to say that the removal of the spleen is
probably the removal of a healthy organ in most cases of ITP.
Let me know if anything I said contradicts any info. that you
have.  This helps rule out some ideas and bring others to the
forefront.  Also let me know if I can be of anymore help.

http://www.niddk.nih.gov/health/hematol/pubs/itp/itp.htm

11. Dr. Rea had me take seratonin to guard against platelet reaction and it seemed to work.  Why?  Seratonin is a major component of platelets?  What is seratonin?

The Details and My Attempt to Decipher  

(In Aplastic Anemia) ... Morphologically, the bone marrow is devoid of hematopoietic (blood cell formation) elements, showing largely fat cells. Flow-cytometry shows that the CD34 (CD34 is a transmembrane glycoprotein constitutively expressed on endothelial cells and on hematopoietic stem cells. This highly O-glycosylated molecule, containing serine and threonine-rich mucin like domains, binds to L-selectin, but its functional capacity in non-lymphatic venules is uncertain. Studies have suggested that CD34 is important in tethering lymphocytes.  - WHAT THE HELL DOES THAT MEAN?????

Mice deficient in CD34 exhibited no detectable abnormalities in postsurgical leukocyte rolling in cremaster venules. Antibodies blocking L-selectin function reduced rolling in CD34 deficient mice suggesting that CD34 lacks major significance as a ligand for L-selectin. The endothelial ligands for L-selectin are currently unknown.)   MY GOD - WHY DON'T THEY SPEAK ENGLISH!!! cell population, which contains the stem cells and the early committed progenitors, is significantly reduced. In vitro colony culture assays suggest profound functional loss of the hematopoietic progenitors, so much so that they are unresponsive even to very high levels of hematopoietic growth factors.  Our bone marrow is deficient in the vital cells that ultimately produce platelets, WBC's and RBC's (progenitors).

Little evidence points to a defective microenvironment as a cause of aplastic anemia. In patients with severe aplastic anemia, the stromal (Connective tissue cells of an organ found in the loose connective tissue) cell function is normal, including growth factor production. Adequate stromal function is implicit in the success of marrow transplantation in aplastic anemia because frequently the stromal elements remain of host origin.

The role of an immune dysfunction was suggested in 1970, when autologous recovery was documented in a patient with aplastic anemia who had failed to engraft after marrow transplantation; Mathe proposed that the immunosuppressive regimen used for conditioning promoted the return of normal marrow function. Subsequently, numerous studies have shown that in approximately 70% of patients with acquired aplastic anemia, immunosuppressive therapy improves marrow function. Immunity is regulated genetically (by immune response genes) and also influenced by environment (eg, nutrition, aging, previous exposure). Although the inciting antigens that breach immune tolerance with subsequent autoimmunity are unknown, human leukocyte antigen (HLA)-DR2 is over-represented among European and American patients with aplastic anemia.

Suppression of hematopoiesis (formation and development of blood cells) likely is mediated by an expanded population of cytotoxic T lymphocytes: cluster of differentiation 8, HLA-DR+ (CTLs: CD8, HLA-DR+), which are detectable in both the blood and bone marrow of patients with aplastic anemia. These cells produce inhibitory cytokines, such as gamma interferon and tumor necrosis factor, which are capable of suppressing progenitor cell growth. (The T lymphocytes are killing the progenitors!) These cytokines suppress hematopoiesis by affecting the mitotic cycle (cell division) and cell killing (apotosis) through induction Fas (Fas is a known inducer of apoptosis and is important in the regulation of several aspects of the immune system, including cytotoxic killing of cells potentially harmful to the organism such as virus-infected or tumor cells.)-mediated apoptosis (Programmed cell death) It also has been shown that these cytokines induce nitric oxide synthase and nitric oxide production by marrow cells, which contributes to immune-mediated cytotoxicity (The phenomenon of target cell destruction by immunologically active effector cells. It may be brought about directly by sensitised T-lymphocytes or by lymphoid or myeloid "killer" cells, or it may be mediated by cytotoxic antibody, cytotoxic factor released by lymphoid cells, or complement) and elimination of hematopoietic cells.

Anemia is a disorder that results in a decrease in the ability of the blood to carry oxygen. Anemia is itself not a diagnosis but merely a sign of underlying disease. The initial classification of anemia is best accomplished by examination of the data from a hematology analyzer and by an examination of the peripheral blood smear. The physician most commonly classifies anemias initially by the instrument's red cell indicies, especially the mean corpuscular volume (MCV). On newer counters, the red cell distribution width (RDW) or red cell morphology index (RCMI) is another useful measurement. The anemia may be microcytic, normocytic, or macrocytic.

Additional Definitions, Glossaries and Revelations

MACROCYTIC ANEMIA

Macrocytic anemias are less commonly encountered than normocytic or microcytic anemias. These anemias may be caused by marrow failure such as aplastic anemia and myelodysplasis, or caused by deficiencies of vitamin B12 or folic acid; or caused by autoimmune hemolysis or cold agglutinins.

Myeloid Stem Cell
Produced by the pluripotent
stem cell. It will eventually
mature into red or white blood
cells or platelets.

Lymphoid Stem Cell
Produced by the pluripotent
stem cell. It will mature into
a T cell or a B cell

Hematopoiesis
The formation and development of
blood cells

Red Blood Cell (Erythrocyte)
Contains haemoglobin which
carries oxygen. Lives for 120
days

Platelets (Thrombocytes)
Helps to control bleeding
by clotting blood at sites of
injury. Lives for 5 - 9 days.

Neutrophil
A white blood cell that
engulfs and kills
bacteria

Macrophage
A white blood cell that
engulfs and kills
bacteria

Monocyte
Travels to areas of
infection where it
becomes a macropahge

Eosinophil
Kills parasitic worms
and removes bacteria

Basophil
Intensifies and hastens
the immune response
to infection

Mast Cell
Similar to a basophil but
found in tissue instead of
blood

B Cell
Produces plasma cells

Plasma Cell
Makes antibodies
which recognize and kill
infections

T Cell
Kills infections directly and
stimulates B cells to produce
antibodies

Myeloblast
Immature cell which
eventually becomes a white
blood cell

B Lymphoblast
Immature B cell

T Lymphoblast
Immature T cell

Monoblast
Immature monocyte

Megakaryocyte
One megakaryocyte
produces 4000 platelets

Megakaryoblast
An immature
megakaryocyte

Proerythroblast
An immature red blood
cell

Myeloblast
Immature cell which
eventually becomes a white
blood cell

Myeloblast
Immature cell which
eventually becomes a white
blood cell

White blood cells, depending
on type of cell, it may live for
a few hours or a few years

Pre B Cell
Immature B cell

Prothymocyte
Immature T cell

Hematocrit

• What is the hematocrit?
• How is the hematocrit measured?
• What is a normal hematocrit?
• What does a low hematocrit mean?
• What does a high hematocrit mean?

What is the hematocrit?

The hematocrit is the proportion, by volume, of the blood that consists of red blood cells. The hematocrit (hct)is expressed as a percentage. For example, an hematocrit of 25% means that there are 25 milliliters of red blood cells in 100 milliliters of blood.

How is the hematocrit measured?

The hematocrit is typically measured from a blood sample by an automated machine that makes several other measurements at the same time. Most of these machines in fact do not directly measure the hematocrit, but instead calculate it based on the determination of the amount of hemoglobin and the average volume of the red blood cells. The hematocrit can also be determined by a manual method using a centrifuge. When a tube of blood is centrifuged, the red cells will be packed into the bottom of the tube. The proportion of red cells to the total blood volume can be visually measured.

What is a normal hematocrit?

The normal ranges for hematocrit are dependent on age and, after adolescence, the sex of the individual. The normal ranges are:

• Newborns: 55-68%
• One (1) week of age: 47-65%
• One (1) month of age: 37-49%
• Three (3) months of age: 30-36%
• One (1) year of age: 29-41%
• Ten (10) years of age: 36-40%
• Adult males: 42-54%
• Adult women: 38-46%

These values may vary slightly between laboratories.

What does a low hematocrit mean?

A low hematocrit is referred to as being anemic. There are many reasons for anemia. Some of the more common reasons are loss of blood (traumatic injury, surgery, bleeding colon cancer), nutritional deficiency (iron, vitamin B12, folate), bone marrow problems (replacement of bone marrow by cancer, suppression by chemotherapy drugs, kidney failure), and abnormal hematocrit (sickle cell anemia).

What does a high hematocrit mean?

Higher than normal hematocrit levels can be seen in people living at high altitudes and in smokers. Dehydration produces a falsely high hematocrit that disappears when proper fluid balance is restored. Some other infrequent causes are lung disease, certain tumors, a disorder of the bone marrow known as polycythemia rubra vera, and abuse of the drug erythropoietin (Epogen) by athletes for blood doping purposes.

What is Apotosis?  (Cell Biology)

Chart below shows what Roche Diagnostics know about apoptosis (physiologically regulated cell suicide).  In Aplastic Anemia, our immune system causes premature apoptosis. Apoptosis is the opposite of necrosis - cell death by accident.  Further information on apoptosis including a clickable version of the chart can be found at http://biochem.roche.com/prodinfo_fst.htm?/apoptosis/

I'm pretty sure that understanding CD34 and how it relates to the immune system, protein status and amino acids is critical to finding the immune system linkage I am looking for, but look at this and tell me what it means!?

ALTERNATE NAMES FOR CD34  

• CD34 [HUGO gene name]
• gp105-120

MAJOR LINKS FOR CD34  

• NCBI LocusLink Record: 947
• Mendelian Inheritance in Man (OMIM): 142230
• SwissProt annotated protein record: P28906

CELLULAR FUNCTION OF CD34 Link to additional info in FORUM  

• Cell-cell adhesion
• Inhibition of hematopoietic differentiation?

DISEASE RELEVANCE OF CD34 AND FUNCTION OF CD34 IN INTACT ANIMAL  

• No abnormality in leukocyte trafficking was detected in the CD34 knock-out mice
• In 1 of 2 reports of CD34 knock-out mice, a decrease in hematopoietic progenitors was found in the knock-out mice
• Utilization of CD34 mAb to quantitate and purify lymphohematopoietic stem / progenitor cells for research and for clinical bone marrow transplantation

• Families in which CD34 is a member
  • CD34-->sialomucin-->Mucins

MOLECULAR STRUCTURE OF CD34  

• A heavily glycosylated type I transmembrane protein. There are two forms of the CD34 protein, resulting from alternative splicing
• The complete extracellular region is present in both forms of CD34
• There is a cysteine-rich repeat (Ig-like domain) in the extracellular region
• The full-length form of CD34 molecule has an intracellular domain, which contains consensus sites for protein kinase C (PKC) phosphorylation, serine and threonine phosphorylation by other kinases, and tyrosine phosphorylation (To date, only serine phosphorylation by has been actually demonstrated)
• The truncated form of CD34 lacks most of the intracellular domain, including many of the potential phosphorylation sites

  	Full-length form	Truncated form
  Full amino sequence	385	328
  Intracellular region	73	16
  Transmembrane	23	23
  Extracellular region	258	258
  Signal sequence	31	31

MOLECULAR MASS OF CD34  

POST-TRANSCRIPTIONAL MODIFICATION OF CD34  

• One species contains exons 1 through 8 and forms the full-length form of CD34
• An alternative splice variant results in the insertion of an additional exon (exon X, 194bp) between exon 7 and 8; this introduces a translational stop codon, which results in the truncated form of CD34 with a shorter cytoplasmic domain
• The transmembrane and extracellular regions of both forms of CD34 are identical

POST-TRANSLATIONAL MODIFICATION OF CD34  

• Beginning at the NH2 terminus, the extracellular domain is heavily N- and
• O-sialoglycosylated
• Serine phosphorylation of the intracellular domain has been demonstrated, and there are potential sites for serine, threonine, and tyrosine phosphorylation

SUBSTRATES FOR CD34   - No information

ENZYMES WHICH MODIFY CD34   - No information

LIGANDS FOR CD34 AND MOLECULES ASSOCIATED WITH CD34  

• Expressed on early lymphohematopoietic stem and progenitor cells, small-vessel endothelial cells, embryonic fibroblasts, and some cells in fetal and adult nervous tissue
• Also, expressed on hematopoietic progenitors derived from fetal yolk sac, embryonic liver, and extra-hepatic embryonic tissues including aorta-associated hematopoietic progenitors in the 5 week human embryo

SELECTION OF OTHER CD34-SPECIFIC REFERENCE MAB  

REVIEWS

1. Krause DS,Fackler MJ,Civin CI,May WS CD34: structure, biology, and clinical utility. Blood 1996 87:1 PubMed

2. Sutherland DR,Keating A The CD34 antigen: structure, biology, and potential clinical applications. J Hematother 1992 1:115 PubMed

PRIMARY CITATIONS

3. Baumheter S,Singer MS,Henzel W,Hemmerich S,Renz M,Rosen SD,Lasky LA Binding of L-selectin to the vascular sialomucin CD34. Science 1993 262:436 PubMed

4. Civin CI,Strauss LC,Brovall C,Fackler MJ,Schwartz JF,Shaper JH Antigenic analysis of hematopoiesis. III. A hematopoietic progenitor cell surface antigen defined by a monoclonal antibody raised against KG- 1a cells. J Immunol 1984 133:157 PubMed

5. Fackler MJ,Krause DS,Smith OM,Civin CI,May WS Full-length but not truncated CD34 inhibits hematopoietic cell differentiation of M1 cells. Blood 1995 85:3040 PubMed

6. Gaudernack, G. and Egeland, T. Leucocyte typing (ed.Schlossman, S.F. et al), pp. 861-4. Oxford University Press, Oxford (1995)

7. Huyhn A,Dommergues M,Izac B,Croisille L,Katz A,Vainchenker W,Coulombel L Characterization of hematopoietic progenitors from human yolk sacs and embryos. Blood 1995 86:4474 PubMed

8. Lin G,Finger E,Gutierrez-Ramos JC Expression of CD34 in endothelial cells, hematopoietic progenitors and nervous cells in fetal and adult mouse tissues. Eur J Immunol 1995 25:1508 PubMed

9. Nakamura Y,Komano H,Nakauchi H Two alternative forms of cDNA encoding CD34. Exp Hematol 1993 21:236 PubMed

10. Perrotti D,Bellon T,Trotta R,Martinez R,Calabretta B A cell proliferation-dependent multiprotein complex NC-3A positively regulates the CD34 promoter via a TCATTT-containing element. Blood 1996 88:3336 PubMed

11. Sato N,Sawada K,Koizumi K,Tarumi T,Ieko M,Yasukouchi T,Yamaguchi M,Takahashi TA,Sekiguchi S,Koike T In vitro expansion of human peripheral blood CD34+ cells. Blood 1993 82:3600 PubMed

12. Satterthwaite AB,Burn TC,Le Beau MM,Tenen DG Structure of the gene encoding CD34, a human hematopoietic stem cell antigen. Genomics 1992 12:788 PubMed

13. Suda J,Sudo T,Ito M,Ohno N,Yamaguchi Y,Suda T Two types of murine CD34 mRNA generated by alternative splicing. Blood 1992 79:2288 PubMed

14. Sutherland DR,Abdullah KM,Cyopick P,Mellors A Cleavage of the cell-surface O-sialoglycoproteins CD34, CD43, CD44, and CD45 by a novel glycoprotease from Pasteurella haemolytica. J Immunol 1992 148:1458 PubMed

15. Tavian M,Coulombel L,Luton D,Clemente HS,Dieterlen-Lievre F,Peault B Aorta-associated CD34+ hematopoietic cells in the early human embryo. Blood 1996 87:67 PubMed

16. Tindle RW,Nichols RA,Chan L,Campana D,Catovsky D,Birnie GD A novel monoclonal antibody BI-3C5 recognises myeloblasts and non-B non- T lymphoblasts in acute leukaemias and CGL blast crises, and reacts with immature cells in normal bone marrow. Leuk Res 1985 9:1 PubMed

WWW RESOURCES

Portions copyright by Garland Press and by the International Workshops on Human Leukocyte Differentiation Antigens; used with permission

Modified 10/14/99   prow@ncbi.nlm.nih.gov

The hallmark of aplastic anemia is the empty bone marrow, and by all measures hematopoiesis is markedly reduced. Not only are the distinctive hematopoietic precursor cells, the young forms of the erythroid and myeloid lineages and megakaryocytes, absent by visual examination of the aspirate and biopsy morphology, but imaging of the marrow spaces of the vertebrae shows uniform replacement with fat. Cells bearing the CD34 antigen, measured by flow cytometric phenotyping and a marker of the progenitor and stem cell compartment, are virtually absent in blood and marrow. Functionally, cells capable of forming erythroid, myeloid, and megakaryocytic colonies in semisolid media are much reduced, and in vitro assays of very primitive, quiescent hematopoietic cells, close to the true stem cell, show a similar consistent and severe deficit⁷. Estimating from these assays, it is likely that patients with aplastic anemia present with pancytopenia when their stem cell and progenitors have fallen to about 1% or less of normal numbers. While some blood cells are produced even at this low level, evidence of the enormous compensatory capacity of the marrow, such a profound deficiency has important qualitative consequences, perhaps reflected in the shortened telomere length measured in granulocytes of patients with aplastic anemia,⁸ compatible with an extremely stressed state of hematopoiesis. A deficient hematopoietic stem cell compartment is replaced in the course of bone marrow transplantation.

The viability and differentiation of blood cell progenitors depend on specific hematopoietic growth factors, mainly produced by the marrow stroma(connecting tissue) However, laboratory studies have generally shown normal function of aplastic anemia patients’ stroma, and the circulating blood levels or in vitro production of almost all cytokines is normal and indeed elevated in the great majority of patients.9 The contention that stromal accessory function is largely normal in aplastic anemia is also supported by the success of bone marrow transplantation, after which many stromal elements remain of host origin, and the general ineffectiveness of growth factor administration in patients with severe disease.

The efficiency of immune system destruction of hematopoiesis is also obvious in animal runt disease and in human transfusion-associated graft-versus-host-disease; in these syndromes, small numbers of alloreactive T cells lead to fatal aplastic anemia.¹¹ A large amount of laboratory data supports the hypothesis that in most patients with acquired aplastic anemia, lymphocytes  (White cell of the blood that are derived from stem cells of the lymphoid series. Two main classes are recognised, T and B lymphocytes, the latter responsible (when activated) for production of antibody, the former subdivided into subsets (helper, suppressor, cytotoxic T-cells) and responsible both for cell-mediated immunity and for stimulating B-cells) are responsible for the destruction of the hematopoietic cell compartment⁹. 

Early experiments showed a suppressive effect of patients’ lymphocytes on hematopoietic colony formation of normal persons and on the patients’ own bone marrow. These cells produced a soluble inhibitory factor that ultimately was identified as -interferon (<protein> A family of glycoproteins proteins derived from human cells which normally has a role in fighting viral infections by preventing virus multiplication in cells. IFN alpha is made by leucocytes and IFN gamma by fibroblasts after viral infection. IFN _ is produced by immune cells after antigen stimulation. IFN _ and _ are also called type I interferons, IFN _ as Type II interferons, more usually classed as cytokines.) cytokine, protein> A variety of naturally occuring polypeptides that are members of the family of cytokines which affect functions of specific cell types and are found in small quantities. They are secreted regulatory proteins produced by lymphocytes, monocytes and various other cell types and are released by cells in response to antigenic and non-antigenic stimuli.

Interleukins are of the larger class of T-cell products, lymphokines which are now more frequently considered as cytokines. The interleukins, of which there are 12 identified to date, modulate inflammation and immunity by regulating growth, mobility and differentiation of lymphoid and other cells. Included among the cytokines are cachectin and lymphotoxin which are now known as tumour necrosis factor-alpha and tumour necrosis factor-beta, respectively.

Interleukin-1 inhibitors are the first well-described proteins involved in the feedback regulation of interleukin activities. The origin and functions of the two most novel cytokines, interleukin-11 and interleukin-12 (also referred to as natural killer cell stimulatory factor), have only recently begun to be understood.

Cytotoxic lymphocyte activation, and most recently the intracellular presence of type 1 cytokines,¹² can be measured by flow cytometric methods in patients’ blood and marrow. The result of this immune process is destructive, with CD34 cell death induced through the Fas-Fas ligand pathway protein> Cell surface transmembrane protein (35 kD) that mediates apoptosis ) and by activation of intracellular pathways leading to cell cycle arrest and the release of nitric oxide. This compound is produced from L arginine by the enzyme nitric oxide synthase. Acts as a potent vasorelaxant via elevation of intracellular cGMP in vascular smooth muscle.

Immunity is local and has been modeled in vitro in cell culture systems in which low concentrations of -interferon are secreted into the marrow microenvironment. In an animal model of aplastic anemia in which bone marrow failure is produced by injection of alloreactive lymphocytes, pancytopenia can be prevented by treatment with a monoclonal antibody to -interferon.¹³ In summary, acquired aplastic anemia appears to share an autoimmune pathophysiology with other human diseases in which T cells effect organ-specific apoptosis of target cells.

Crux of the issue may be determining what foods/antigens/other incitents may impact cells bearing the CD34 antigen either positively or negatively  and/or understanding what is happening genetically.

 That's bubba's hypotheses for now.

amino acid

A class of organic molecules that containing an amino group and can combine in linear arrays to form proteins in living organisms.

There are twenty common amino acids: alanine, arginine, aspargine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine.

They are key components in all living things from which proteins are synthesised by formation of peptide bonds during ribosomal translation of messenger RNA.

All the amino acids have the L configuration, except glycine which is not optically active. Other amino acids occurring in proteins, such as hydroxyproline in collagen, are formed by post translational enzymatic modification of amino acid residues in polypeptide chains.

There are also several important amino acids, such as the neurotransmitter y aminobutyric acid, that have no relation to proteins.

Amino acids can now be produced by biotechnology in bulk using fermentation and biotransformation.

Acronym: AA

What is the linkage between CD34, proteins, cytokines, amino acids and interkeukin?  Is it reasonable or advantageous to increase amino acids and/or proteins to balance the immune response or is that working against me?

In the last century, both our understanding of the origin of aplastic anemia and the definitive and supportive treatment of the individual patient have improved enormously. Aplastic anemia in most cases is immunologically mediated, sharing pathophysiologic mechanisms with other autoimmune diseases. Unfortunately, stem cell destruction may be quite advanced by the time the patient presents with pancytopenia. Replacement of hematopoietic tissue by either stem cell transplantation or suppression of the pathologic immune response to allow recovery of the patient’s own marrow are effective. Improvements in methods for monitoring hematopoiesis and immune system activity and especially in the application of immunomodulatory drugs should be of practical benefit in the clinic. Major research questions remain about the nature of inciting antigens and the determinant of the aberrant immune response, as well as the fundamental pathophysiologic relationship among aplasia, dysplasia, and paroxysmal nocturnal hemoglobinuria.

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October 4, 2001 Update

The more I read and research, the more I am convinced that AA should be treated like other autoimmune diseases.  Most of them are described in very similar manners and treatments are also very similar.  So I will continue to research autoimmune diseases in general and look for environment and clinical approaches towards healing the immune system.

October 8, 2001 Update

Pretty sure that I am onto something - CD34 presence is significantly reduced in patients with AA.  CD34 is somehow linked to threonin, cystine and some other amino acid (i'm too tired to look it up).  I am low in all of the aforementioned amino acids so will take supplements.  Let's see what happens.  There is a part of me that believes it couldn't be that simple or one of the brilliant researchers at NIH would have tried it--- but what the heck - i have nothing to lose.  my alternative is to continue to get transplants or do a BMT.  I am definitely going to exhaust every environmental/natural approach i can find.

October 18, 2001 - Kenton Joins the Fight

Thank God for Kenton - He is my nephew and has a strong background in biochemistry. He currently works for a major pharmaceutical company and is helping to decipher the CD34 mystery and some of the other stuff that I am having a hard time understanding.  Here is what he has contributed so far:

CD34 Research Relative to Amino Acids

My analysis and fundamental questions posed to Kenton

1. Is the addition of tons of vitamins and supplements (C, A, B12, Folic Acid, E, Multi, Selenium, Arginine, Taurine, Cystine, Threonine, etc.) really worth the effort or am I just switching from medicine to vitamins.

2. Explain and understand the CD34 threonine linkage. My threonine is low, threonine is component of CD34?  CD34 is significantly reduced in AA patients.  I lose it afar that - is there something that can be done, or I can do, or genetic/stem cell researchers may find that will alleviate this CD34 deficiency?

“Something is destroying my hematopoeitic cells before they mature.
Interesting that L-selectin binds cd34 and is seen as a homing receptor for lymphocytes.  It is only supposed to bind endothelial cells (possibly for destruction) and it is not supposed to bind to hematopoeitic cd34.”

 My understanding goes like this:

Aplastic Anemia essentially says that one or both of the following are happening:

1. My stem cells are not properly creating the essential blood cells (RBC, WBC, Platelets)

2. The progenitor cells are being attached by my lymphocytes (killer T Cells) because they are being recognized as malformed by my immune system

In all the "stuff" out there, I have narrowed it down to these statements:

"Morphologically, the bone marrow is devoid of hematopoeitic (blood cell creation) elements, showing largely fat cells...CD34 cell population which contains the stem cells and committed progenitors (the parent cells in the bone marrow for red cells, white cells and megakaryocytes. The most primitive hematopoietic progenitor cells are multipotent and give rise to the progenitor cells for red cells, white cells and megakaryocytes).  In vitro colony assays suggest profound functional loss of the hematopoietic progenitors, so much so that they are unresponsive even to very high levels of hematopoietic growth factors."

" Immunity is regulated genetically (by immune response genes) and also influenced by environment (nutrition, aging, previous exposure)...human leukocyte antigen (HLA-DR2) is over represented among patients with Aplastic Anemia."

"The result of this immune process is destructive, with CD34 cell death induced through the Fas-Fas ligand pathway and by activation pathways leading to cell cycle arrest and the release of nitric oxide."

"Suppression of hematopoiesis likely is mediated by an expanded population of cytotoxic T lymphocytes (killer cells): cluster of differentiation 8, HLS-DR+ (CTLs: CD8, HLA-Dr+), which are detectable in both the blood and bone marrow of patients with Aplastic Anemia.  These cells produce inhibitory cytokines, such as gamma interferon and tumor necrosis factor, which are capable of progenitor cell growth.  These cytokines suppress hematopoiesis by affecting the mitotic cycle and cell killing through induction Fas-mediated apoptosis (regulated cell destruction).  It also has been shown that these cytokines induce nitric oxide synthase and nitric oxide production by marrow cells, which contributes to immune mediated cytotoxicity and elimination of hematopoietic cells."

"Cells bearing the CD34 antigen are virtually absent in bone and marrow (of AA patients).  Functionally, cells capable of forming erythroid, myeloid, and megakaryocytes colonies in semisolid media are much reduced."

Then following the track I mentioned before, I went off and tried to learn more about CD34 and formed the linkage with Threonine, Arginine and Cystine.

This information certainly seems to point to an effort toward "balancing the immune system" which is Dr. Rea's claim to fame and I appear to making that happen, but counts are still not rising.  I feel much stronger and have eliminated a lot of toxins, but counts crashed last week.

Kenton's Edited Response

There is a relationship between CD34 and low amino acids.  CD34 is a glycoprotein (protein that contains a part that is a carb) that spans the outer membrane of cells.  Proteins (and glycoproteins are made up of constituent groups called amino acids.  These amino acids are linked by peptide bonds (chemical bonds between the carboxyl and amino groups of amino acids) the low levels of both may be related. 

It appears from a chemical structure of cd34 that it may made up primarily of the amino acids of which I am deficient (Threonine, Arginine and Cystine). Maybe my low levels of cd34 are due to low levels of amino acids and my body cannot synthesize the protein because of this but I believe it more likely that the low levels of amino acids are due to low levels of cd34.  I don't believe that my body is having problems producing cells with cd34.  It is possible that the cd34 protein or something that binds to the protein is marking the cells for destruction.  Thus destroying the amino acids. 

Increasing intake of amino acids is worth a shot, but it may be detrimental to kidneys (excess protein).  Most of the amino acids that are low are non-essential.  My body should be able to produce these by itself.  If my body was having problems producing these amino acids I would expect more health problems than anemia.  The other essential amino acids are brought in from extraneous sources.  Since I am low in both essential and non-essential I should look in other areas for the problem. 

CD 34 is a membrane protein.  It has a part that extends well outside of the cell and a part that extends inside of the cell.  Many membrane proteins are transport proteins that act to bring "stuff" inside or outside of cells. Seems like the actual structure of cd34 is in dispute.  Can't seem to find anything definitive.  I do see that there are possible phosphorylation sights for all of the amino acids I am deficient in.  (Phosphorylation is an energy favorable reaction) not important but in the fact that it shows me that the extra cellular tail probably is made up of a lot of the aminos I specified.  (Only
one definitive though) Cd34 is also possibly related in allot of cell-to-cell binding.  Found primarily in hematopoietic cell lines.  Cell to cell binding such
as clotting (platelets may have an unusually high number of cd34 binding sites or more active cd34 binding sites.

Something is destroying my hematopoeitic cells before they mature. Interesting that L-selectin binds cd34 and is seen as a homing receptor for lymphocytes.  It is only
supposed to bind endothelial cells (possibly for destruction) and it is not supposed to bind to hematopoietic cd34.  Maybe it is binding hema cd34 in
my system.  

All right cd34 and threonine.  Okay let’s start from the beginning.  Again, cd34 is a protein.  Proteins are made up of amino acids.  Amino acids share a similar chemical
structure, most notably that they all have two "ends" one carboxyl end and one amino end (that’s where they get their name from).

Chemically I do not have to know what constitutes these groups as long as I
recognize that they both have differences in charge (think magnets likes repel opposites attract) These amino acids are linked (bonded) together by peptide bonds between the two chemical groups of different amino acids. 

Amino acids linked by peptide bonds.  These linked amino acids are proteins.  These
proteins can actually include chemical structures that are not altogether protein in nature.  We are going to focus on glycoproteins.  Glycoproteins have somewhere
within their structure a chemical component that is carbohydrate in nature (not an amino acid but a structure that is organic in nature (primarily Carbon)and is usually made up of Carbon Oxygen and Hydrogen. 

They are usually primary energy sources for cells.)  In other words this protein has been attached to a carbohydrate and has formed a stable product. (Not predisposed to being broken apart into its constituent amino acid and carbohydrate groups)  This is cd34.  A
glycoprotein.  CD34 is a membrane protein.  This means that it is found attached to the outer membrane of cells.

They are usually attached between the two phosholipid bilayers of the cells. Cells have many membrane proteins.  Some only extend above the membrane (in contact only with stuff outside of the cell).  Others extend only into the inside of the cells.  Others extend both inside AND outside of the cell.  These are transmembrane proteins. (they extend across the cellular membrane)  This is the type of protein that is cd34.  I would guess that the carbohydrate portion of this glycoprotein is the section that actually spans the membrane (the charge of the amino acids would not bond there very well. 

A phospolipid does not like charge.  It has a charge on its phosphate end but this is attached to a long hydrocarbon tail that lacks charge.  Because this tail does not like the charges of the amino acid it would not want the amino acid to be located very close to the tails.  This is where the neutral (more or less) carbohydrate section could come into place.  It acts as a bridge that connects to proteins on each side of the cell membrane.  Because this is stable it is labeled a glycoprotein. 

Many membrane proteins allow particles (charged particles (ions etc.) that would normally not be allowed across) into the cells. Others actually just send signals into the cell when the proper receptor is stimulated.  I'm not sure what cd34 is used for yet but I do have some ideas.  Funny thing is that the exact structure of cd34 is in dispute.  There are definitely two conformations.  One has a section that extends into the cell only a short distance.  The other extends into the cell a much longer distance.  It sounds like the extracellular portion of the protein is the same for both
conformations (structures).  Because of the similar structure of cell membranes between all cells it could be very difficult to differentiate between the cells.

This is where proteins come into play.  Each cell has a different membrane protein profile.  This is the mechanism that the immune system uses to attack
invading cells.  If it does not have proteins that are seen as being the same as the organism the cell is marked for destruction.  Okay that’s it for now but I
still have to explain threonines potential role in this.

Good Link from Melanie - Center for Blood Research

http://cbr.med.harvard.edu/investigators/index.html

A large number of human diseases of unknown origin, but mostly involving immune reactions against the patients’ own tissues, show associations with specific genes of the major histocompatibility complex (MHC) region of chromosome 6. Our laboratory studies the genetics of immune function in humans, particularly within the MHC. Specifically, we investigate the relationship between genetic differences in the human MHC with differences in the immune function of a variety of "white blood cells," or leukocytes.

November 10, 2001

Update from Kenton on CD34 Fas-Fas Ligand research with a great animation of the apoptosis process:

I'm sending you this link because it visually explains the Fas-Fas Ligand pathway of cell apotosis, thought to play a role in the destruction of the Megakaryocytes.  I've been going through alot of the postings on your web site.  You've done a very good job with research and presentation.  You would make a good scientist. 

Basically if this is the system of cell destruction then it indicates that the Fas molecule is being incorrectly triggered by Fas Ligand and destroyed. Now the question is what is marking your cells for this interaction to occur (look at the viral epitope in the diagram, this is what is marking that cell for destruction).  It could be that your Megakaryocytes are displaying a surface antigen that is foreign when they are produced.   If this is the case it would basically indicate that it is not your immune system that is malfunctioning.  It is simply doing what it has been instructed to do since you were born.  Destroy cells that are properly marked.  This is a problem with the expression of antigens on the surface of your Megakaryocytes.  Has little to do with a malfunction in your immune system. 

The other possibility that I had seen was that your immune system was accidentally reading your "self" cells as "non-self" (foreign bodies).  This would mean that there was a problem with the receptor sites on your immune cells.  They were destroying normal cells that had normal types and amounts of binding sites.  This is more like the over active immune system theory.  (Instead of the viral epitope in the diagram place one of your own cell surface proteins.   cd34?)  This can also incorporate the Fas-FasL apoptotic pathway.  There are however probably several different types of AA that occur because of either of the above reasons. Anyway I hope you are feeling better soon.  Here is the site.  It does not deal directly with AA but it gives an overview of how and why this system is activated. MHC-1  Major histocompatibility complex-1 TCR  T-cell receptor CD8  cell protein on T-lymphocytes

http://www.cat.cc.md.us/courses/bio141/lecguide/unit3/apofas.html  - something weird about this link - cut and paste the url into your browser

April 16, 2002 Restart

After giving up in frustration and getting back to a somewhat normal life, I will restart with cd34 research.  I believe the fundamental issue is the following:

What is causing my bone marrow to produce hypocellular aspirations and/or what is causing the resulting cells to be marked with the cd34  marker.  The highlighted discussion from Kenton above is the start, now I need to go on and flesh this information out into something that I can understand. The discussion below is from the AA Facts Page - Time to go back to the beginning and challenge all the assumptions to date.

A bone marrow biopsy is performed in addition to the aspiration so that the cellularity may be assessed both qualitatively and quantitatively. In aplastic anemia, these specimens are hypocellular. Aspirations alone may appear hypocellular owing to technical reasons (eg, dilution with peripheral blood), or they may look hypercellular because of areas of focal residual hematopoiesis. A core biopsy gives a better idea of cellularity; the specimen is considered hypocellular if it is less than 30% cellular in individuals younger than 60 years or less than 20% in those older than 60 years. A relative or absolute increase in mast cells may be observed around the hypoplastic spicules. A proportion of marrow lymphocytes greater than 70% has been correlated with poor prognosis in aplastic anemia. Some dyserythropoiesis with megaloblastosis may be seen in aplastic anemia.

Gathering data points as an old boss used to say:

cellularity

The degree, quality, or condition of cells that are present.

cellular immune theory

A concept, put forth by Elie Metchnikoff, that cells, not antibodies, were responsible for the immune response of an organism.

(05 Mar 2000)

Pick it up here - My BMB site is hurting enough so I can't really think anymore.

Note - The term hypocellular is not in the medical dictionary so who is it that has the authority to invent a word which is at the root of our disease?

May 10, 2002 Discussion on Stem Cell Research with Nephew Kenton a Biologist

Hi Kent,

Hi Uncle Bruce, how are you? Don't worry about bothering me. I don't mind reading over research papers for you. Just sometimes I might not be able to respond immediately. The article you sent me is actually very vague in content. I am trying to access the actual report or a more techincal press release. The homeobox genes, such as HOXB4, serve to encode transcription factors involved in the process of normal development and differentiation of cells and organs. This means that they do control the production of different types of cells from stem cells. They use the term "supercharge" but it sounds like the cells are induced to divide at an accelerated rate by the introduction of this gene. Thus they are able to use small numbers of stem cells to produce large numbers of stem cells that in turn become large numbers of blood cells which can be used to rebuild a damaged immune system. When ready for human trials the patient would have their cells destroyed by irradiation, the use of drugs (cyclosporin) or combinations thereof (depending on what needed to be wiped out). The stem cells generated in the lab could then be introduced into the patient where they would quickly differentiate into the various blood cells. The paper says nothing about whether or not repeated injections of the genetically altered stem cells would be needed. As the original stem cells that were introduced die off or differentiate, will new ones be produced to take there place? If so these new cells that are produced will be the genetically modified form with HOBX4 inserted into them. Will these cells continue to proliferate at the rate observed in the laboratory? This could lead to an overabundance of stem cells and a resulting overabundance of blood cells. They would need a way to "turn off" the gene or at least regulate it. Sometimes this is simple other times it isn't. The prospect of inserting gentically altered material into patients is controversial. There is no way of knowing how this material will interact in living systems. Mutations to the gene may occur (As the article says some homeobox genes can produce flies with 4 wings, legs on their heads). But sofar this along with fetal tissue research offer the best possibilities for AA. Honestly both are a long way off. This because it has just been performed and fetal research because of the contoversy involved. The gene interacions for this procedure must be mapped out and explained before this will go anywere and then more effective animal trials would have to be performed before they even thought of trying this in a human clinical trial. Chances are they would prefer to use products derived from the expression of HOXB4 instead of the actual insertion of the gene. This would delay the developments even further as the interaction of each of the products with the cells was researched. As I said the paper was vague so if I find anything else out I will e-mail it to you. If you have anymore questions let me know. Do you understand how fetal tissue research could be used to produce stem cells? Do you want to know how different genes can be turned on and off by various chemical signals in the lab or in the body?

http://www.cap.org/captoday/archive/2002/platelet_disorders_feature.html -

Back in the research mood starting August  18, 2002 - What exactly is going on with my blood cells?

October 4, 2002 - How are hemophilia, AA, AIDS, MDS, and other such diseases related ?  What are the common threads?  How are they dissimilar?

What is going on - I saw an article on gene therapy for hemophilia - dopes is apply?

Hemophilia defined:  

haemophilia

<disease, haematology> A haemorrhagic diathesis occurring in two main forms:

1. Haemophilia A (classic haemophilia, factor VIII deficiency), an X linked disorder due to deficiency of coagulation factor VIII.

2. Haemophilia B (factor IX deficiency, Christmas disease), also X linked, due to deficiency of coagulation factor IX.

Both forms are determined by a mutant gene near the telomere of the long arm of the X chromosome (Xq), but a different loci and are characterised by subcutaneous and intramuscular haemorrhages, bleeding from the mouth, gums, lips and tongue, haematuria and haemarthroses.

(18 Nov 1997)

Hemophilia is considered an ideal disease for gene therapy because it's caused by a single malfunctioning gene, and only a small increase in clotting factor in the bloodstream could provide great medical benefits

AIDS - Trying to understand the linkage

There appear to be lots of similarities - What are the differences?

Important note: Information in this article was accurate in 1995. The state of the art may have changed since the publication date.



An autopsy case of acquired immune deficiency syndrome (AIDS) with preceding aplastic anemia.

Pathol Int. 1994 Dec;44(12):850-6. Unique Identifier : AIDSLINE MED/95170923
Nagasaki M; Harada T; Torii I; Nakano A; Furuya H; Tanaka J; Hirai K; Morikawa S; Department of Pathology, Shimane Medical University, Izumo,; Japan.

Copyright ©1990, 2000. ÆGIS. All materials appearing on ÆGIS are protected by copyright as a collective work or compilation under U.S. copyright and other laws and are the property of ÆGIS, or the party credited as the provider of the content. Feedback/Contact Us.

Other Related Diseases:

Myelodysplasia Acute Leukemia Myelofibrosis Lymphoma Multiple Myeloma Hairy cell leukemia B12 deficiency Folic Acid Deficiency

So, what are the similarities and differences with:

Lupus, Aids, Hemophilia, Non Hodgkins Lymphoma, Hodgkins, Leukemia, MDS and other autoimmune related disorders?

I absolutely agree on the elimination of supplements and trying to get everything naturally. I believe the amino acids/proteins are central to our problems but I would rather get them naturally. I am about to drop all vitamins and supplements. The research is primarily aimed at trying to gain a fuller understanding of why the cells are being malformed in the first place. I have a good feeling that we are doing the right things as far as getting the immune system in balance but still think we need to figure out why the cells are being marked for destruction in the first place. I am also trying to regroup and assemble all of my notes along with yours and others to find the rosary stone. I absolutely agree on the issue of more people sharing their strategies . There are so many thread to follow it is hard to stay focused, but we will prevail!

Choline

Introduction

Movement of a limb is achieved by exciting a muscle so that it contracts. Muscles do not contract of their own accord, they rely on messages from the brain via nerves.

How do nerves tell muscles to contract?

A signal is transmitted from the brain to the end of the nerve attached to the muscle which it wishes to contract. There is a small space between the end of the nerve and the muscle and it is across this neuromuscular space that the nerve pushes small quantities of a chemical called acetylcholine. When sufficient acetylcholine attaches to the outer surface of the muscle cell, the muscle cell becomes excited and contracts. Should you run out of acetylcholine then the muscles will not be able to contract, even if the they are adequately stocked with energy (ATP).

Physiologists reckon that acetylcholine is actually broken down inside the neuromuscular junctions during prolonged exercise. Nerve cells then "grab" the choline floating by in the blood, using it to make new acetylcholine and as a result, your blood choline levels start decreasing. Naturally, if your choline levels fall too far, acetylcholine production can come to a relative standstill, and your nerve cells will simply refuse to stimulate your muscles.

What is the function of Choline?

Choline is a vitamin like compound which is an essential part of the human diet as it is used buy the body to produce acetylcholine. Without choline, acetylcholine cannot be produced and the the body cannot function normally. Choline is also an extremely important structural element of cells, especially cell membranes, and is essential for the process of breaking down fat for energy.

Choline and the 'Wall'

When you run a race like the 5km, 10km or half-marathon the choline concentrations remain sufficient. It appears that your choline levels drop dramatically only when you run a marathon or exercise continuously for approximately two hours or more. Some exercise scientists believe that this drop in choline is behind the devastating fatigue which strikes near the end of a marathon - referred to, by many marathon runners, as the 'Wall'.

Some scientists reason that choline supplements if taken at the right time and in the right amount might help the nervous system continue to stimulate muscle cells and keep you striding toward the marathon finish line at your desired pace.

Where does choline come from?

Choline is available in foods such as: liver, cauliflower, soybeans, spinach, lettuce, nuts, and eggs . The bottom line is that even a choline rich natural diet probably would not prevent the drop in blood choline levels which happen after 20 miles or so of marathon running.

For a marathon, how much and when?

The right amount is probably about 2.5 grams, swallowed about an hour before your marathon begins . This additional dosage of choline in your blood may begin to fall three hours after you've taken it (e.g., two hours into your marathon), so it makes sense to take another 2.5 gram dose at the 10 to 13 mile point of your race.

Any side effects?

Choline is perfectly safe to take, the only potential problem being an occasional bout of diarrhoea or some pretty foul flatulence.

Supplements

'Boston Sports Supplement' contains carbohydrate and electrolytes along with choline and is sold as a powder which you mix with water.

Sources of choline in our daily diet are: liver, cauliflower, soybeans, spinach, lettuce, nuts, eggs and wheat germ.

Associated Pages

The following Sports Coach pages should be read in conjunction with this page.

• Ergogenic Aids
• Supplements

Associated Web Sites

The following web sites contain more information on this topic:

• None at present

Associated Journals & Books

The following journals and books contain more information on this topic:

• Peak Performance - Issue 73 (page 7)

Help

If you would like any further details or help on this topic then please email me .

CD34 is a transmembrane glycoprotein constitutively expressed on endothelial cells and on hematopoietic stem cells. This highly O-glycosylated molecule, containing serine and threonine-rich mucin like domains, binds to L-selectin, but