J. Govshiov
Department of Physics, University of Texas, Austin, TX.
March 10 1997
Let me first say, in case some of you after reading this article, nod their head and say in bewilderment "this is the most incredible bits of non sense I have ever read", this could be for a reason. I am not a native student to the arts of the flesh. Nor was I ever trained at a recognizable institution except the library. My only knowledge is from self training that arose from a passion to understand the processes involved in Central Nervous System (CNS) regeneration. My original and founded training comes from the realm of physics. As such, I will greatly appreciate any advice and critic any professional researcher would like to share with me about the content and accuracy of the ideas presented.
Let me add, that I see my roll and propose as the young child that shouts "Thay King is Naked". Or in this case "The implication of Spinal Cord Regeneration (SCR) and cloning."
Since the early phases of physiology, man has grown to understand that the heart is a great pump but the brain is the "brain" behind human operation. Our entire complexity, our soul is imbedded some how in the complex jelly-mater of the brain and the s pinal cord, the Central Nervous System (CNS). Later scientific observation found that the local behavior of the CNS is electromagnetic (EM) in nature and carried out by different ion circuit. These ions also function as carriers of different peptides and other chemicals needed for different body functions. Through the balance of these two delicate physical phenomena nature has created life. More appropriately its the old action principle, that says than nature always chooses the extrimal point of the act ion in any system. Basically natural processes are those that are carried at just the right energy.
Two scientific developments of this recent age give rise to an operation that will someday change the face (literally) of the human-kind. This is, rather complicated but still possible with the aid of technology, available and researched at the down of the millennium. This paper will try to outline the procedures necessary to perform this operation. some of the are still being studied at laboratories around the world. This operation, put in over simplified terms, is the regeneration of the brain of ag ing bodies on a brand-new cloned body.
Now that we have stated the goal we can formulate a way to it. I delay discussion of the enormous benefits that will arise, and the applications to "good" genetic engineering to the closure of this article.
First and far most important for this experiment to succeed is to forget the old ways of physiological research. The "lets butcher it and see what happens" method will with no doubt result in the unsuccessfulness out come of this experiment. Rather w e try to save its life. As to the number of species in each set of experiment I think the best starting point would be seven middle aged rats. Where each operation will be considered unique. For notation sake, lets call the rats by {P,Q,R,S,T,X,Y} and ass ociate a number to each. For example X1 would be the original and X2 would be the clone.
Upon dissection of the spinal cord drastic physiological effects take place. Axoplasm in large quantities is purred out of the lesion. Experimentally we find that the greatest chance of recovery occurs in those injuries that suffered the least trauma. I.e. Where the amount of "wasted" fluid is low and the transection is not mutilated. So its plausible that we'll perform an operation that will be the least traumatic to the spinal cord.
The critical factor, determining the destructiveness of the lesion is probably the amount of axoplasm removed. It is probable that generation of collateral sprouts is dependent on nerve cell bodies for synthesis of new axoplasm. Also for big axopla sm loss we see big RNA reduction.
It is because those factors that I suggest having both X1 and X2 in a state of deep hypothermia and performing the surgery while the body is in this state. An obvious and very important benefit of this would be the elimination of axoplasm loss. Of cour se given that the dissected spine will be securely tucked in place before raising temperature In addition, the place should be kept under cool temperature at the early phases of the healing process. At least for the first week.
Another benefit, is that in deep hypothermia we reduce CNS activity to a minimum. The living, or should i say dying being, CNS's doesn't even "know" that it has been transected. It is only at the wakening stages of the operation that the dissected spin al-cord starts noticing the lesion. It is during this stages that we must make sure :
A : the two parts (the cloned spinal and the original brain) are securely attached and coated by some non-rejective degradable material. B : the lesion surface is coated with Schwann cell's. It reasonable to assume that #Schwann cells needed = (the cross-section area of the white matter in the cord)/(the av. Area of an unmeylented axon).It is very reasonable to assume that opening up the synthetic covering of the lesion or messing with it will result in poor connection and thus should be avoided. Still if remyelation is the goal, we must supply a fresh supply of Schwann cells to the lesion daily, especially at the later stages of recovery. I.e. After the first week. This is a tricky procedure and as such should be performed under careful planing and outlining the appropriate tasks to be carried out. The problem of inserting Schwann c ells to the lesion surface is left for the technical development.
We now, from observation, observe that at the site of the transection there is a lot of lysosomes which under certain circumstances can cause autolysis of healthy fibers in the vicinity.
A question arises : could we suck the lysossmes from the lesion and apply Schwann cells later on. It seems that the destruction occurring is the first step the body takes in the rode regeneration (maybe as a mean of cleaning up). The problem is that in mo st studied cases the body does not have enough resources to finish the processes and so is left in a worse shape then at the time of transection. Regeneration is a hard enough job without the bodies enthusiasm to "kill in order to re- grow".From experimental studies we find that the stump in axon growth is the myelination of the axon. Physiologically axon is the path through which ions travel. This electromagnetic pulse is built by a fine arraignment of the Schwann cell on the axon tube. On the axon, the Schwann cells wrap and form the nod of revenuer. Through this "separator between Schwann cells" occurs the electric conducting. In lame terms one can say that positive ions enters the axon thus creating a sudden shift at the field inside the axon. This creates an attractor for the negative ions which stream there. In turn, this attracts another positive ion and we have an EM wave.
The important thing to remember is that this is a stable system. We have tow effects that are occurring : 1) the Schwann cell disabling positive ion penetration through most of the cell surface. 2) the positive ion disables growth of Schwann cell in short rings on the axon surface.Now we can better understand what happens at the later stages of the spinal lesion. Once there is an actual dislocation in the spine. There can be no electric pulse coming through from the lesion location and maybe more importantly there can be no cond uction through the entire length of that portion of axon tube. In any case we see that the Schwann cells, especially on the exposed parts, start to myelinate "every thing in there path". Perhaps this is a means for the for the body to prevent further axoplasm loss. So in order to stabilize growth, we need to sustain a natural, stable, chemical environment. This can be achieved by lab generated EM waves because of the net charge of the chemicals involved.
We now come to the "path finding" by the axon. I thought and dealt with this problem quiet a lot, how could -under trillions of possibilities- the axon find its way back? Lets say that after we transplant the spine of Mr. X to his appropriate clone, an d lets assume for a minute that some how all the tissues connect!
Still, our body is made out of different components. Say the arm and the leg. They both connect through the spinal cord to the brain. We know that all commands come from the brain. It would be very bad if the PR guy had to explain, why Mr. X says he wants to drink coffee while kicking the reporter. Or in other words we must have mo re then mere connections, we must have the right connections!
Luckily again for us, a picture of a cross section of the axon in the CNS will help to understand how they will form the right. And I'm sure it would.
Locking at (fig - 1) (TO BE ADED)- a cross section of the axon, we see that the axon shape varies from tube to tube. In some cases they even differ in topological class. (From: Axonal specification of Schwann cell expression and myelination, P.S. Spanc er and H.J. Wienberg in Waxman : physiology and pathobiology of axon). It is pretty obvious to me that this patterns are the way by which axon membrane finds its path back to its original location. Moreover it is pretty clear that this forms could be trac ed back to our DNA.
Experimentally we find strong support that this is the case, in Roger Spary's (I might have the wrong spelling). In these experiments Spary and his colleagues rotated the eye of frog in various angle and exchanged between the left and the right eye. T hey found out, say for a rotation by 180 degree, the frog would associate images from above with down movement and vice versa. This means that the axons reconnected to their original location. The same result was obtained for eye interchange.
Another extremely interesting topic is the connection of wave form and propagating through various shapes. With our physical knowledge and computation it should be possible. In later work I will study the effect of shape variation on the propagating na ture of the pulse.
I know very little about these subjects. Attempts to find Information on cloning and incubating abilities did not succeed, especially On studies performed in the United States. As though research does not Exist or is covered up quite successfully. I kn ow from discussions with several biologist that some of the engineering capabilities that will be described later, are present within our present capabilities, but I'm not sure as to what extent.
We basically need to be able to incubate a clone. And to actually fit the original (grown) brain in the growing skull. If any body knows of research being done on growth stimulation? This information can prove highly valuable. Notice that for experime nts to be done on rats this factor is not as important because of the relative rapid maturation of the rat brain as compared to humans.
As the starting point for the surgical operation I would take the later 2/3 of the embryo development. It is during this time that the brain is most developing. Also it will enable the regeneration to take place in a system that is already growing. What we need:
1. Clone an embryo from X1 (from the same genetical code of the original - this is very important for a couple of reasons: i) As was pointed out earlier the axon will find the right place to hook because of its shape. This in turn is connected to the genetic code of the individual. ii) As in any operation there is the possibility of rejection and because of the nature of this operation it seems to that it will play an even bigger role. iii) The brain and the spinal cord constituted the emerging embryo. The chances of this operation ever being done between two genetically distinct specimens seems very low.First we need to study the nature of the pulse at the designated lesion. We need to know the amplitude and the frequency of the pulse at the cross-section. I hope I would be able to calculate these numbers from general conditions. The key issue here i s the axial length of the Schwann cells *(the wave length). The axon of the CNS is relatively homogenous so if we assume constant velocity we can find the frequency we need to generate. The problem of amplitude seems less obvious and will be addressed in later work.
Viewing the axon microscopically we see that there is a stable balance between the ions and the myelin sheath. On periodic intervals there is no coating and through these points the ions penetrate the cell, creating local imbalances in the EM field. I t is reasoned here that this is a two-way process. i.e. That the myelin prevents ion to enter the axon AND that the ion penetration prevents the total myelination of the axon. So it would prove highly beneficial to produce the same pulse in the disconnect ed axon.
There are a couple of possibilities. The first would be to use "wiring" already present in the clone brain. Experimental evidence shows that certain brain mater improves the generating abilities of the axon. We can drive electrical pulses from the out side through this mater to the lesion. The other possibility is to use a short metal cylinder around the spinal cord in order to create an EM pulse inside the spinal. Careful consideration must be taken because of the permitivity nature of the covering su rface. I am not sure which of this will succeed (if any) but the second seems more appealing because of its non-evasive nature. Hopefully this will give the adjacent axon with enough of its natural environment in order to suppress demyelination.
At the beginning of the third portion of pregnancy we begin the actual transfer. We start by bringing both X1 and X2 to a state of deep hypothermia. The reasoning behind this was described earlier. In general we wish to prevent large scale degradation of the CNS. Now we come to the actually transfer. The crucial point here I think is where to make the final cut. i.e. The final connection. I'm unaware of actual experiments on the correlation between the place of lesion and the regeneration capabilities of the CNS.
I suggest that in the initial 7 rat sample we vary The transection site. The suggestions are: 1. At the tuber cinereum. Separating the pons from the temporal lobe and Keeping the oculomotor nerve. 1.1 Keeping the cerebellum of X1. 1.2 Keeping the cerebellum of X2. 2. After the mammillary bodies. Again separating the pons from the Temporal lobe. 2.1 Keeping the cerebellum of X1. 2.2 Keeping the cerebellum of X2. 3. An horizontal cut, just below the cranial nerves VI,VII,VII. Separating at the connection between the medulla oblongata and the pons. 3.1 Keeping the cerebellum of X1. 3.2 Keeping the cerebellum of X2. 4. An horizontal cut at the bottom of the medulla oblongata. Connecting at the median aperture This time only keeping X1 cerebellum. *All cases connection the cerebellum at the superior penduncles.This ordering of the lesion location. Going downward along the Brain stem and spinal cord is intended. Although at first it seems that the last two locations (# 3,4) should prove more successful, I think the first two have better chances for a couple of reasons. All research done suggests the time of the embryonic formation as the time of most successful SCR. The total axon paths damaged in the first two location would be the least because of the small cross section of the damaged axon. The smaller cr oss section would aid the axon reconstruction at the site of lesion.
Notice that in method #2 we can vary the lesion in X1 and X2 and make both surfaces of the connection with the same cross section. This is sure to aid the regeneration process.
Remember that both X1 and X2 are under a state of deep hypothermia, so there should not be a considerable loss of axoplasm and other necessities. For case #1 I suggest the use organic glue to connect X2 Mammillary bodies and X2 tuber cinereum with X1 's temporal lobe. For case #2 one could use a cone shaped, non-rejective polymer, attached to X2 Mid brain and anchored to X1's temporal lobe.
For cases #3 and #4, Insert the two ends into two cylinders. The first would go on X2 spinal. It should be at the exact diameter of X2 spinal cord. On X1 spinal end I would put a shorter cylinder again at X1 cord diameter. Finally we should connect th em together. This will serve for several functions,
1. Provide support for the healing of the cut. The same concept of a fracture. 2. Help stop swelling. 3. HOLD THE AXON ALIGNED VERTICALLY AS IT SHOULD. Notice that if there is a significant age difference between X1 and X2, hence the diameter of the two cylinders may vary. Thus special consideration must be applied to the connection and covering of the cylinders.We must transfer the brain in its entirety. It is my opinion that we should transform the old cerebellum with it. The strongest argument I feel is that the cerebellum coordinates most of our every-day functions. At first the mind performs this operatio ns in the developing new born. As the brain gets used to performing them they are moved to the cerebellum. This priceless knowledge could prove life saving to our awakening rat. But I feel that this subject should be experimented.
We're basically implanting the old brain on the new spinal cord. This operation, if possible will reduce many of the obstacles standing on our path.
Now is the time to start the "de freeze" process on our system. Notice that still the brain is on life support. i.e. Blood circulation is maintained from the outside. It is expected that the fresh supply of blood and the higher pressure will induce mor e rapid growth in the embryo. These life support should be disconnected and the final connection of blood vessels to the brain should take place only at a stage where the born body of X2 can safely able to sustain enough blood circulation. This can be don e in stages.
After the initial shock it is time to apply Schwann cells and other growth stimulator to all connection sites, these could come directly from the embryonic brain. Earlier I discussed only the lesion to the spinal cord or the brain stem. It was assume d that other tissue including peripheral nerves will regenerate. These sites could still benefit from a little hand. Hopefully by the time X2's body is able to sustain life support to the brain all nerve and axon connection have formed. We are almost fini shed.
I say almost because we are left with the problem of the size of the scull versus the size of the brain. One thing we might try is to keep the scull cap with X1's brain and connect it to X2's opened scull. I'm not sure if this procedure is possible. I would appreciate any information on this mater.
Other questions that arise are: The possibilities of prolonging the embryo and the brain, in the incubator, in order to generate a bigger clone. The ability to manipulate one aspect in the embryo development, say the size of the skull. The ability of the brain to survive in embryonic fluid - experiments suggest that the spinal fluid an embryonic fluid are very much related.
This is a very complicated operation, there is no doubt. But it is not a science fiction story. Most of the procedures outlined have been performed in the past. Some steps to successfully complete this operation are still missing. Some of these arise, I hope, from my lack of knowledge. But some of them are serious problems that need to be handled with seriousness. Most of these problems are engineerical in nature. Luckily the human engineering abilities are very much evolved.
The successful out come of this procedure will radically change the way we view the medical process. Deficiencies that proved fatal in the past would become curable. I give a short list of the possibilities opened by this procedure. I'm sure that with little imagination one can find numerous more.
1. Considerably extending human (and animal) life span. To a point unimagined in the past. We are not at the steps of immortality yet, but this will bring us allot closer. 2. Cure to many of today's incurable diseases. We can interact with our RNA. We can manipulate our own genetic code! For instance we can remove from the clone recognizable "bad genes". A patient grown with a Genetically Determined Disease (GDD). Could be re grown with that genetic deficiency removed. And there are even simpler cases.We can correct pregnancy deficiencies as well. Say a pregnant mother drinks heavily during the pregnancy. In many such cases, the baby is born with incurable physiological deformations. Cloning a new host in a stable environment (in those cases where the brain was not effected) will cure the deformation.
Notice that in both these cases the operation should prove more successful, or at least easier to perform. There isn't such a drastic size difference between the host and the brain, so we don't have a drastic problem at the skull. Second, both the hos t and the brain are at their growing stage, as I pointed out earlier this should prove beneficial.
3. Any injury inflicted on the body, in which the brain was not harmed. Consider for instance a practical procedure to the battle-field, in which the first aid treatment to a causality is by putting him in a freeze bag. This bag already exists for various other purposes. Assuming no brain damage this bag can be evacuated to the nearest lab. Their a clone may be grown and the soldier just might eventually have a normal life. 4. Technically it would be possible to choose the gender of the clone! All we need to do is to change one chromosome. This is realizable. Thus it would be possible for (the male) Mr. X to bare children and for the female Mrs. X to impregnate him! I'm neit her transvestite nor gay. But this possibility I'm sure will appeal to many. It does to me at a later cycle.And the list goes on and on. The outcomes and possibilities of this procedure stretch the limits of our imagination.
To the late Louis and Pedros. For I'm sure that without their
Tender love and care, I would not have come to this.
To him that will make this successful.
to All.
If any body has comments, suggestions or corrections to this document, I would beg of them to feel obliged to send me an email on the mater. Thanks in advance.
On Remyelination by Schwann Cells,
1. Blight A.R. and Wise Young (1989) Central axon in injured cat spinal cord recover electrophysiological function following remyelination by Schwann cells. J. Neurol. sci., 91: 15-34 2. Blakemore W.F. Remyalination of CNS axon by Schwann cells transplanted from the sciatic nerve. Nature, 266: 68-69 3. Blakemore W.F. and Crang A.J. (1985) The use of cultured autologous Scwann cells to remyelinate areas of persistent demyelination in the CNS. J. Neurol. Sci., 70: 207-223 4. Harrison B.M (1979) Remyelination by cells introduced into a stable demyelinating lesion in the CNS. J. Neurol. Sci., 46: 63-81On Growth Mediators:
1. Bregman B.S. et al. (1995) Recovery from spinal cord injury by antibodies to neurite growth inhibitors. Nature, 378: 498-499 2. Iwasaki Y. et al. (1991) Trophic effect of angiotensin II, Vasopressin and other peptides on the cultured ventral spinal cord of rat enbryo. J. Neurol. Sci., 103: 151-155On Recovery Aids:
1. Blight A.R. and Gruner J.A. (1987) Augmentation by 4-aminopyridine of vestibulospinal free fall responses in chronic spinal-injured cats. J. Neurol. Sci., 82: 145-159 2. Bostock H. et al. (1981) The effects of 4-aminopyridine and terathylammonium ions on normal and demyelinated mammalian nerve fibres. J. Physiol. 313: 301-315 3. Smith K.J. et al. (1981) The restoration of conduction by central remyalination. Brain 104: 383-404Evidence of Conduction by Anatomical Continuity:
1. Dimitrijevic M.R. (1988) Residual motor function in spinal cord injury. In: Waxman S.G. (Ed.), Functional Recovery in Neurological Diseas, Raven Press, New York, pp. 139-155 A Good Reference: 1. Seil F.J. (Ed.), Neural Regeneration and Transplantation, A.R Liss Publ., New York. 2. Waxman S.G. (Ed.), Physiology and Pathobiology of Axons, Raven Press, New York.On E&M Theory:
1. Jackson J.D., Classical Electrodynamics, John Wiley & Sons, New York.