ENZO has Patents for binding sites on nucleic acids:

   In essence, the building blocks for DNA, and RNA comprise amino acids which offer preferred binding sites. ENZO has Patented these sites and shown that a certain embodiment, a molecule, can bind reliably and present some for of detectable element. Again, in one preferred embodiment, a phospholuminescent molecule is presented for what amounts to photo detection. All non-radioactive, of course! In yet another Patent, ENZO solved the problem of sticky molecules. Molecules presented in "chains" like hair on a surface, tend to clump or stick. This "quenches" the signal, when a signal is applied to the system (flash of light, for example). ENZO applied like charge to the molecules used for detection, via the choice selection of those molecules (many different molecules can serve the function).
   As such, by way of specific selection, the molecules all contain like charge and they maintain their equidistance and juxtaposition. As photonic energy is directed toward these molecules, the phospholuminescent molecule at the head of the chain, produces a unique signal back to the photo detector. Equate this to a field of sunflowers, no light present, a flash of light from space and an observer in space, next to the source of the flash, looking for the yellow flash back representing the heads of the sunflowers. If all sunflowers are facing the same direction, the system detects the presence of the sunflowers with 100% accuracy, but only because these "flowers" affix only to a targeted field with their highly selective roots, if and only if the binding between root and field is perfect. If not, the flowers would wash away and no such effect would be visible from space. By selection, the stems all contain like charge and thus they present the head of the flower in relative like angle, making the molecules more detectable from a great distance. This analogy works well to illustrate why probing of a target cannot be accomplished with only one molecule of the target and the target is first amplified (into a field of identical targets). Then when probed, the response is unequivocally registered as positive or negative and the response is 100.0000000000 accurate. (as in, extremely accurate). Only then, is a "net genomic solution" to a "net genomic problem", rendered accurate and reliable. Imagine how important this is to sequencing and desequencing, and the accuracy of the work done?
 

   This technology launched the industrialization of DNA, RNA and even Proteomics, as tracking Proteins down to their source involves probing of the RNA/DNA logical pathways. To be meaningful, the process must be accurate and must be reliable.

   (((((BRENNER's first major point))))) moving on...

   Genetic Antisense comes next, as the first set of developments led ENZO to the next logical evolutionary step. If ENZO is uniquely endowed to discover the pathways with its own unique tools, what next?
   Regulation up or down, of those genes which occur in nature! What better way than learning to place "Antisense" Genes, or in the alternative, the placement of "Sense" Genes, or both? Brenner discusses the naked concept. An Antisense Gene does not touch or even necessarily exist within proximity of its targeted Sense Gene. Instead, it merely compliments the target, with its own production of micRNA (Message Inverse Complimentary RNA). Whatever the target gene produces the Antisense Gene paired to it by way of structure, nullifies through hybridization. Gene A produces mRNA X, Antisense Gene A' produces micRNA X', where X and X' bind with extreme efficiency. Outside a living cell, where a protein was once observed to be produced, the placement of the correct antisense gene halts all production of this protein, even though the original gene giving rise to the protein is still present in the nucleus, untouched, unaltered and producing its mRNA. ENZO can regulate anything a cell does! ENZO can add a function to a cell! With Gene Editing, ENZO can make minor corrections, apply minor enhancements, or perhaps shunt a gene altogether.

   Genetic Antisense is far superior to Oligonucleotide because, in order to work at all, Genetic Antisense must be permanently added to a given cell's nucleus. Oligo is applied periodically and thus poses great and permanent, insurmountable issues. Oligonucleotide delivery, concentration and reliability will remain as problems. The larger the molecule and the more complex, the harder to deliver and to maintain in numbers within cells, where the work needs to be completed. Brenner makes this distinction. IMO, if you want to fix a highly complex machine, you can either build a work-around outside the core of the machine or go into the machine and fix it according to its nature. Everyone knows the permanent fix is the desirable fix. Oligo is inferior to Genetic Antisense because it equates the same as the prior illustration.
   More importantly, like any machine, everything effects everything else. Genetic Antisense forces the scientist to perfect the placement of the genes and trial run the final model to perfection. Oligos exist in cytoplasm only and do not reveal their long term effect as to rise and fall of concentration and elimination.
   A Genetic Antisense placement transcribes at a certain and known pace and the placement of the antisense gene is permanent, affixed according to promotor and terminator, all known commodities and all yielding predictable behavioral models. CONTROL over the chromosomal content is in question here.
   Genetic Antisense wins out. Last analogy, IMO, this is like fixing software by utilizing the original compiler to write a more perfect program, versus, running an entirely different external program to try and fix a problem after it occurs. Oligo is reactive by nature, Genetic Antisense is proactive by nature.  Brenner understands this distinction.

Brenner notes that Oligo can be highly toxic, applied outside to inside, as is necessary for any higher multicellular organism. But if produced by the nucleus, can be perfectly acceptable.

   Brenner concludes (Read this with tears in your eyes!!!)>“Therefor an approach such as ENZO’s ought to find application to any disease, because the approach focuses on the source of the problem, rather than its expression”. WOW! HE GETS IT!

   What does he mean? I think it is quite simple. In an overly simplistic model, one can think of evolution as lining up problems and organisms. The organisms that fail, fail according to the problems. Those that survive, survive do so because of mysterious differences, perhaps superiority to the organisms which tried and failed. But science has taken this observation to its extreme. The molecules that formed in early primordial oceans, leading to the living cell, the multicellular organism and essentially, to us, did so according to rules. Rules exist at all levels, molecular, polypeptide, structure, cell, organelle, organ, organism. His point? Disease is a disruption of the rules at some level. ENZO is looking at the GERMANE level. The ultimate and precursory level upon which a simple FIX can defeat a given virus, pathogen, bacterium, fungi, protein deficiency or overproduction. Guess what? ANYTHING YOU WANT A CELL TO DO, WITHIN NATURE, ENZO CAN MAKE THE CELL PERFORM!!!! That is his point. Creative use of this (these) convention(s), allows a scientist to teach a human cell to produce inhuman inverse complimentary viral micRNA’s. To the human cell, it is of no consequence whatsoever. To the virus attempting to gain entry and replicate, it is fatal. Apply three versions, and a certain virus may not be able to mutate or replicate or even slightly recombine within a human cell, altered as such. 9801-230 or the Genetic Antisense product HGTV-43, represents just such an advantage, added to CD34+ immune system precursor cells, targeted to HIV-1. (or HIV-2)

   IT WORKS!

   HURRAY! Moving on...

   Gene Editing. Brenner is astonished and so am I! ENZO scientists must have been studying the retroviral mechanisms, exactly how a virus adds its DNA to existing human chromosomes. They thought “why can’t we write a virus which attacks the human chromosome in much the same manner, but instead of teaching the chromosome to make copies of the virus, we instead provide instruction for replacing a defective sequence or adding a sequence for enhancement or even a meaningless shunt of information to shut down the target?” It can be done. It was done. Teaching that a TNA or “Triple Helix” state can be provoked, that the provocation can be accomplished to allow the DNA sequence to naturally select a more desirable sequence and to see that sequence bind; this is what ENZO has accomplished. The mechanisms deployed by virus’ to perpetuate their numbers by forcing human cells into submission have been harnessed. ENZO can reverse a viral effect. Rewrite the virus’ coding. Write coding to fix natural mutations or provide fixes to undesirable sequence edits. Enhance a gene. Repair a gene. Turn off a gene from inside the gene. Erase a portion of a gene. Indeed, the hyper typewriter effect with automatic erasure is a good analogy. I would say, a typewriter that remembers the dynamic structure of a whole paragraph (equating paragraph to whole gene) so at the end of the paragraph, the writer can say, “oops, I see three mistakes, or four, or five etc... and can enter the edits at that juncture, automatically fixing all cited characters by scrubbing out the old characters and inputting the new, without failure or flaw.
   Amazing! GENE EDITING. ENZO literally has genies in their bottles. Make a wish?

Brenner talks about ENZO Probes for identifying disease presence, absence and of course, all with perfect accuracy. “Gene Analysis” such as “Abnormality Detection” or “Infectious Disease Diagnostics” will all come of age through these innovations.

   Brenner mentions the “PathoGene” Product line which identifies which cell is infected and where within the cell, the infection has taken place.

   Brenner says a “second generation” is just about complete. Testing will be automated and currently includes about 45 pathologic targets. This will render Culturing techniques obsolete, creating lots of space in Farmingdale for an indoor ice rink for the kiddies! But better still, ENZO will not be leasing any more space anytime soon. ENZO’s complete base of technology would actually fit inside their headquarters with plenty of room to spare. It would not even fill one office, in theory. Things are getting smaller and more profound, day by day.

   Talks about market sizing and potential.

   Talks about recent Patent wins.

   Covers recent events, in reverse chronology. The mouse model does indeed, host human hepatacytes, and the abstract to be delivered at AASLD does discuss the suppression of human liver cancer TUMORS and CELLS! I strongly suspect human patients utilizing EHT-899 and who had Hepatacellular carcinoma, have seen suppression of their cancer, a fact EASILY proven by classic observation of tumor size and growth progression. If chemo is applied and tumors are readily reduced, EHT-899 will be there to hold the cancer growth in place, rendering the cancer in FULL REMISSION. Theory for now, fact on Oct 27 2000, I suspect.

   Investment thesis:

   HIV/AIDs market size only in US explored at a rate of $20,000 per patient. 18.5 Billion. My note: No one else has a d--- thing coming. Nothing. Zip. If 9801-230 works, ENZO will get this whole market. Any drug with even an inkling of an opportunity to ultimately compete, would need many, many years to pre-clinical
trial, human trial, pass through the FDA and so forth. Remember also, there are other immunodeficiencies such as SCID, which remain to tackle. This model represents one which can displace ablative processes. The only need for ablation is, in my view, a partial to prompt the body to fill the void and to harvest a large number of Hematopoietic Stem Cells, thus solving two problems in one therapeutic action. (Take limited bone marrow, transduce precursors, put them back, no real mass ablation). However, remember this: If a more advanced AIDs condition yields damaged bone marrow stromal cell structures and no precursors to speak of, Bone Marrow Transplant will be necessary. In a late stage AIDs condition, I wonder if there is enough of the old immune system left to mount any defense? BMT’s are not done for the AIDs condition right now, because HIV is still present and the process is essentially foiled by this fact. Now ENZO has a means to protect the transplanted tissues and cells. The scales have been tipped. I am wondering, if you transplant a viable marrow and cellular colony from a compatible donor, does a late stage AIDs patient need rejection meds or not? Normally, they are needed because even when you ablate, plenty of immune system cells remain which will attack the transplanted immune system. In the case of a late stage AIDs patient, there is little left of their immune system. I have to wonder if the anti-rejection drugs can be left behind? If so, the transplant, having been transfected with HGTV-43 will IMMEDIATELY show engraftment, growth, CD4+ mature cells in Peripheral Blood and so forth.

   Most importantly, the remaining organs and organelles which assist in maturation will finally prove (as research supports already) the rest of the human body is left undamaged by HIV and these organs are still functional. This points strongly to the presence of HIV mRNA in the cytoplasm of immune system cells as the KEY culprit in causing the immune system to produce surges of CD4+ T-Cells none of which mature and all of which exhibit a 26 day life cycle instead of the full 90 days. If this is so, a miraculous salvation of the immune system and of course, of many patients, will take place. The only remaining issue is to find a bone marrow donor and deal with any and all secondary issues promptly. There is great hope here. Remember, with SCID children, bone marrow and immune system transplants were successfully completed from a sibling t the patient. As Dr. Cohn noted, “only a small starter colony was required to restore an entire immune system in these children”. No we have to remember, the Thymus is physically huge in childhood and relatively benign in adulthood. But equally, the Thymus still processes in adulthood and the back-up organs are still present. Apparently, the spleen processes T-Cells and the Lymphatic system also processes T-Cells. There is scientific evidence to allow one to project that HGTV-43 is going to change someone’s life for the better, very shortly. In HIV+ Patients, the transition is slow because there is no mechanism to tell the infected cells or for that matter, the Patient’s existing
immune system, “die now, get out of the way”. If there were, the small starter colony would quickly fill the void according to natural hormonal signals produced by the body. The body thinks it has suffered a would and has lost blood content. This is the natural response. Remember the use of leeches and blood letting? Perhaps the observation Drs. of long ago were making, was the body’s direct response to create new cells. Maybe there was a modest medicinal effect. Most certainly the technique was not well understood and overused. But in some cases, it could be effective. Purge some cells to prompt the creation of fresh new cells, from reservoirs deep within bone marrow structures which are not yet infected by the illness in question. Surging fresh new cells into the system could help, but not if a Patient is already weakened and unable to produce new cells as a matter of health and vitality level, pre bloodletting. As I said, a misunderstood practice.   HEP-B market discussed. 2 Billion people with infection.  Brenner says: Easily established 4.2 Billion Dollar market.

   Here, in a minor sense, Brenner erred. He incorrectly states “efficacy out to 20 weeks shown”. Correct by reading the chart ENZO provided. Incorrect if you read the Tolerization model from ENZO’s homepage or simply call and ask Dr. Engelhardt. Tolerization is intended to be a permanent effect. In the animal models, tolerization was applied but then removed. The effect has lasted years. The Patient “202” shown at the UBS conference had EHT removed at week 20. But it is now logically week 56. How do I know this? Grant made in January. That has to be post week 8, where EHT shows its effect. Add in the elapsed weeks. Call Dr. Engelhardt and ask “Is EHT-899 intended to be permanent after week 20”. I know the answer is yes and he can answer the question as it was part of the publicly disclosed materials of the past several years. The one could ask “if Patient 202 has subsequently seen any degradation of the effect, you would have to report this, would you not?” I know the answer is also yes, as the Tolerization model hangs its hat on PERMANENCE. Hence, the effect is permanent. This does not mean that the surrogate markers will not fluctuate, but it does mean EHT-899 need no longer be applied. In theory, HEP-B may clear with time in the treated individuals.

   I hope, if ENZO sees HEP-B viral loads begin to drop from the level established at week 10 (Carrier) and sees natural deterioration of HEP-B’s numbers, they would report this also. it is very important to HEP-B pathology. That the constant condition leading to veremia, is prompted by the immune system to liver reaction and when the highly specific molecular targeting is turned off, HEP-B loses a vital advantage. Constant inflammation of the liver could serve as a harboring shield for HEP-B and when removed, the immune system may well eliminate HEP-B as it does in most patients. Brenner does not note Tolerization model is the perfect HEP-B vaccination.

   The pricing models include limited detail and the summary simply states $111 in just two quarters (current plus 2 more). In general terms, by April, 2001, by MY calculation. The rest is a heap of numbers for the pencil and eraser gang.

   However, the Appendices are huge and contain vast amounts of info. I am impressed by Brenner’s grasp of HIV Pathology. I hope they see a few factors to Gene Therapy, Stem Cell Therapy and ENZO’s approach which I feel are very, very important to consider.

   1) HGTV-43 is not a chromosomal altering application. It works as a discrete loop without effecting the current U-1 region it mimics.

   2) Inside the treated cell, there is NO integration of HIV RNA/DNA and thus NO provirus in the cytoplasm. This factor is proving to be the KEY to understanding HIV’s greatest weapon against the human immune system. It also explains why drugs can never work against HIV. HIV will mutate and will replicate in the presence of drugs. Drugs can never be concentrated highly enough to be effective inside cells. The HGTV-43 approach attacks HIV when it is molecularly exposed, inside cells. It halts HIV 100% and allows 0% integration. It allows 0% mutation. These are key to understanding Genetic Antisense and the applications which will emerge from the model and its protective Patents. BRENNER WAS UNABLE TO ASCRIBE A VALUE TO THIS. Only a $ 600+ target stemming from success with the current applications. I SAY, IF HE SAYS $ 600, $ 2000 IS POSSIBLE, IF THE COMPANY SHOWS IT INTENDS TO AGGRESSIVELY ADD SEVERAL DISEASES TO THE LIST. LEVERAGE TOLERIZATION AND ANTISENSE GENE THERAPEUTICS.

   Agricultural applications and Bioprocessing applications are mentioned. No value ascribed to ENZO from these important areas. But they are there for the taking...

   Appendix B is a very comprehensive study of HIV pathogenesis. Very impressive. Was this provided to him by ENZO? Who cares. It is factual and that is what matters. INTEGRATION is the viral step ENZO seems to have halted with HGTV-43. That is the net result of my read. This makes the living cell act as a roach motel in the strict sense of the term. HIV keeps coming in, unfolding, reverse transcribing and BANG, no integration. A perfect model for the problem at hand. a) Stop HIV from harming the immune system. b) Collect it and destroy it. HGTV-43 does both.

   There are discussions of HEP and Gene Chips. Lastly, a hasty discussion of Proteomics.

   ENZO will be there, in each field, making its mark upon the state of the art in the sciences which it elects to pursue.

   Sincerely,

   Larry Glaser