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Death, Aging, Rejuvenation
Aging, Death, Rejuvenation   2004

I would like to raise a question: Is Rejuvenation possible and how to approach it?
This is just my own opinion about possible procedures. I used certain techniques based on a theory.
As strange as
it sounds some of those techniques and procedures might actually worked.

There are several hundreds theories of aging. New and old ones. Different authors classify them
differently.

I would divide all those theories into 2 groups :

1. Theories of Deterioration from  external cause. "Wear and tear". Theory of free radicals damage
is most fashionable at present time. Antioxidants are considered a cure. Benefits of caloric
restriction  are often explained by that theory. The theory has good objective support in scientific
literature.
Other theories of this group included changes of bacteria in the gut, radiation and accumulation of
mutations during life of an individual, etc. - many more of those theories  were discussed in the XX
century.
2. Internal clock - theories.


I would divide the "internal clock" theories into 2 more subgroups:
1.  Internal clock on the level of cell. Telomeres shortening is the most discussed in literature at
present.
2.  Internal clock on the level of organism. Growth hormone changes was a recent example of
discussion.

Authors are usually very cautious and objective in scientific literature. Telomeres are usually
discussed in connection with "immortal" cancer cells.
Caloric restriction is discussed as a mean of longevity (not rejuvenation per se). Level of growth
hormone (GH) or insulin-like growth factor drops in aged persons.

Mass media speculates wider: telomeres restoration -  possible immortal life,
growth hormone will rejuvenate you, etc.

Original scientific studies usually do not speculate about this.
In no way for example drop of GH level in elderly means that just giving it back will make you
young again.

Scientific literature rarely discuss "Rejuvenation" as it is. Authors mostly talk about "longevity". This
is very different story. Longevity in a wheelchair is very distinct from a youth playing soccer.
One proven method to reach longevity is caloric restriction. Often they say it is the only proven
technique. Using antioxidants as a method to prolong life is in the same category. To put simple, the
Theory says: nutrition produce products of oxidation - peroxides, etc. These products damage
DNA,
proteins, etc. Cell ages. Caloric restriction switches metabolic pathways. There are less free
radicals.
Hence longevity.  Antioxidants work in similar way - they remove free radicals. Another speculation
is
that caloric restriction starting at young age delays reproductive age in animals and this slows down
biological clocks and prolongs life until an environment with high nutrition supply is found.
Animals look small and undeveloped.

Caloric restriction significantly (25%-100% and more) prolonged life in worms, flies, spiders, rats,
mice, etc. Experiments on monkeys and humans are  under way and will take decades.

Well, it is all good  in experiments and theories. Some argue: why in this case prisoners of
concentration camps do not live longer. Another  example came from "Scientific American" - a
person had caloric restriction of something 20 years or more - no effect - he just looks like very thin
undernourished person of 50 years. Examples of opposite opinion also exist (tribes or  groups with
low food consumption or high antioxidants consumption have more centenarians than general
population).
Maybe you really need to start at 10 years of age, look malnourished and weak for the whole life
and live to 100 years, getting to your puberty at 25.
Antioxidants also bring many controversy. Vitamin E did not show many benefits though it was a big
hope for prevention of many diseases of old age.
And so on.  Today there are big trials that disprove the theory. Tomorrow - great experiments that
confirm the theory, at least partially. Many big trials and experiments are published  in leading
journals - Science, Nature, PNAS, New England JM, JAMA, etc.

One bias for caloric restriction experiment in my opinion is following.
Rats live in cage 30cmX60cm for their whole life. There are at least five rats or 20 mice. They live
couple years and die. During the experiments they are celebrities when they live 5 years - some
lucky ones. The Food is a balanced mixture of nutrition. Caloric restriction is - 60% of their regular
meal. They practically don't move - there is no space in cage.
Mostly they sleep whole day. Sometime they fight and mate.
They live  in those conditions at Yale, at Med U of South Carolina, at Russian State Medical
University and I think everywhere else.
To compare our fellow humans we would place 10 people on area of 20 square metres (200 sq.
feet) for 50 years in a row, feeding them with what they want but all the same - let say junk food
from McDonalds.

I remember  my cat (who was partially wild) stole meat and ate until it started to vomit. He repeated
it many times when he was able to steal some steak or whatever. The same happens in wild - lions
have maybe one successful hunt out of ten. When they catch a prey, they eat like crazy. Much more
than they can digest at the moment. So it looks like there is an instinct. Eat as much as possible.
This is not a joke - 60% of people in western countries are overweight.
Abundant high calorie food and lack of activity. It does not seem that for these people any internal
mechanism restricts food consumption.
Applying this back to rats, we can see that what is considered normal consumption - "ad lib" -
maybe actually big overfeeding of the animals in these cages. In this case 60% of caloric restriction
would be just what rats need, just what the  calories they spend in lazy, uneventful daily life.
So called "control", "normal" rats - that fed as they usually fed might be compared for fat
overweight humans. They consume junk food. They prone to the bunch of diseases of obesity and
low activity. Heart diseases, strokes, variety of cancers, arthritis, etc.
In this case all  the hype about caloric restriction would just be brought to the business of mere
balancing diet and activity in so called "calorie-restricted animals". I haven't seen any discussion of
this problem.
I might be wrong. This is why I use caloric restriction for myself.
I did not work with worms and spiders. But my guess would be that they all are also in artificial
standardized conditions. "Control" worms might be also overfed.
To put rats into bigger cages (to increase activity in control group and balance calories and
catabolism) would be incredibly expensive.
Even at present conditions because of "animal lovers" and bunch of other regulations and
considerations (e.g. sterile conditions), the  price of animal housing is sky-high.

You would say animals in wild  would live longer because they balance activity and consumption.
Well, recently I read that life span of animals  in wild is shorter than animals  in captivity.  Correct
me if I am wrong: pandas  live 15-20 years in  nature
and 20-30 in zoos, bears 20-25 in nature and 30-40 in captivity. Civilization has certain benefits -
vaccinations, good medical care, more or less good hygiene.  So just moving into wild conditions
wouldn't make  you younger or allow to live longer.


Now, to discuss rejuvenation procedures, I would need to talk about aging first. In this case
Rejuvenation could be considered as reversal of aging (deterioration).  Longevity  is different story
- it is prolonged life span. Rejuvenation supposedly should lead to longevity. But longevity is not
equal to Rejuvenation.
Aging leads to Death eventually. It is easier to discuss from that end.


All  written here is just plain speculation, take it with a grain  of salt.

------------------------

Death



Death. Why it happens? Why do we die? Why do animals die? Why do plants die?
What is the need of this?
In my opinion mechanism  of death was selected during Evolution.

From the position of Darwin's theory of Natural Selection everything, that benefits survival of
species and gives advantage in Natural Selection, is preserved in following generations.

Improvements were often left unchanged from the moment of life appearance. Though there could
be other ways, certain mechanisms  were accidentally
selected. These improvements are reproduced in the genome of more  complex species.
Billions of years ago, according to the theory of evolution, chemicals randomly organized themselves
into a self-replicating molecule.

Lightings and UV-radiation helped to create first organic molecules.
This phenomenon is reproducible in a lab. The experiments were described in 1950-60.


First self-replicating molecules were probably  RNA.  First enzymes were probably RNA -
enzymes. Then proteins, DNA  and more complex lipid molecules and polysaccharide came to
the scene.


Death as it is did not exist at the stage of Primeval Soup .  Sure some organic molecules
were destroyed , some new created. But in general it was still that swirling and bubbling primary
broth  - quasi  alive in our understanding as a mixture of biochemical reactions.

Everything in the evolution was build  from the previous  blocks selected sometime by  accident.

Appearance of  lipid membranes allows to compartmentalize the primary broth and create first cells.

At the cellular stage we could already talk about the Death. Cell is destroyed, membrane is broken,
everything leaked out. This is the Death.

Content of cell inside lipid membrane is irradiated. Process of crazy molecular swirling is messed up
by free radicals irreversibly. This is also signs of Death.

Yet at the cellular stage  we  cannot talk about  Aging. Death at this stage is accidental, not
programed.

Organic molecules may age (oxidation, conjugation, etc.) and cell would die.  But damaged
molecule are repaired or synthesized fresh usually. Hence, no good reason for a single cell organism
to age. Irreparable  damage from external cause leads to Death, not Aging. This is accidental death.
For multicellular organisms, there is a parental organism that ages and eventually dies after next
generation is born. For mono-cellular organism, there is parental organism that divides and becomes
the next generation. There is no Aging leading to death of parental organism.

Mono-cellular organisms are practically immortal in a right environment.

Mechanism of division was selected during evolution. Cell has volume (3-D). Surface membrane is
measured in square units (2-D). Growing beyond limits cause inadequate supply of nutrition from
environment. Division solves the problem.

Microbes, bacteria are immortal. Some divide every 20 minutes. In a an hour they multiply 8 times.

Tumour cells divide slower. They are eukaryote. Most aggressive divide once a day.
There is no need for Aging.

They would die if you do not feed them. They die when you  kill them with undiluted bleach in a
flask.
Otherwise they grow unstoppable. No aging.


Multicellular organism  supposedly  has several control  mechanisms to prevent excessive growth
and division, to kill an extra cell. Apoptosis, programmed death, is used.

Cancer  cells often loose the control mechanisms.

A scientist from Yale once pointed out to me that we can not say these cells are immortal. Maybe
they divide and parental cell dies.
Indeed.  We do not follow  the fate of every individual cell during experiments.  They should give
more children cells than parental cells die. Otherwise there would not be the multiplication.
It is a possible scenario. We do not follow the  fate of individual bacteria as well. Maybe they
actually undergo aging.

From the other hand  some experiments suggest  that new cells
contain roughly half of the parent cell after division. So it is not Death or
Aging. Next generations  contain 1/4, 1/8, 1/16 and so on  of the parent cell. Any mechanism that
is more complex than simple division would cause disadvantage in evolution and would be quickly
eliminated.
On the level of mono-cellular organism neither  Aging nor Apoptosis make a lot of sense. Though
Nature often have hidden reasons.

Flick's phenomenon and telomeres shortening make sense for a multicellular organism. Cells of
multicellular organism do age. They stop dividing after 70 divisions or so.

A multi-cell organism has clear cut advantage over a mono-cell.

Why would mechanism of Death   selected for multicellular organisms?
Scenario could be the following.

Big organism have more advantages  compare to smaller one - stability, protection, etc.
There are not so many  natural enemies for  whale or elephant  because of their size.
Large animals could be brought down by bacteria, viruses, small parasites, and lack of available
food. Rare predator attack whale or elephant.

No wonder that  evolution was going  into the direction of bigger animals  - just look  at giant
bones of dinosaurs and you will feel some appreciation.

Millions years ago not only Dinosaurs  but also insects or crustacean were enormous. Cockroaches,
spiders or dragon flies with size of one metre  (3 feet).
Going back in time we find shells of enormous molluscs.
Many things imply that bigger animal with unstoppable growth have a lot of advantages compare to
small ones. New species often evolve in their size. An example is growth acceleration in  humans.
Females prefer taller mates that lead to taller children.
Shorter plants usually cannot compete with the taller forms in the wild. A short mutant in a patch of
tall plants would be shaded out


Mechanism of  reproduction was also selected during the evolution. Any new animal, be it a mouse
or a human, starts life as a single cell. That cell differentiates and develops into the complete animal.
It assures that a rare advantageous variant sequence of DNA will be rapidly propagated by natural
selection. It is much easier to reproduce and maybe get some  new genes or favourable mutations
starting from a single cell than starting from a whole multicellular organism. Functions of most genes
will have been
optimized by random point mutation and selection in any given species. Signalling between cells
during the development process ensure that everything ends up in the right place. Tiny changes in
these Signalling processes can have very large effects on the resulting animal. Genome, with at most
forty-sixty  thousands  genes, is able to specify the creation of a human body containing trillions of
cells, billions of carefully wired neurons and hundreds of different cell types all amazingly sculpted
into organs as diverse as the liver and the brain. This is why mechanism of proliferation of  fertilized
oocytes was selected.

Hence, having younger (smaller) and older (bigger) animals make sense.

But there is the problem:  adult animals (or plants) consume all the nutrition around and there is not
much left for next generation.

Then it is why  mechanism of Death was selected as a mean for next generation to establish.

Imagine the following:

First, mechanism of proliferation  was selected . If mono-cellular organism  does not have
mechanism of reproduction it does not produce offspring with possible  favourable mutations.
Let say there are primitive bacteria spontaneously created in primeval soup. Just few of them. They
consume nutrition, they live happily, practically immortal but they do not divide. Then set of enzymes
appeared during mutations. The set allows bacteria to split. Synthesis of the new set is
disadvantageous. It requires additional energy and nutrition. But advantage of splitting  allows new
mutants immediately overgrow non-proliferating cells.
Selection is really cutthroat.

Bacteria growing in a tube represent  mini evolution. Any bacteria that fit for the environment are
also selected on the ground of simplicity. In a toxic environment (antibiotics)  mutant bacteria having
a new mechanism of neutralisation would survive. But the more simple bacteria will eventually
overgrow complicated  ones. Unless complication gives  big advantage  that outweigh   the benefits
of simplicity.
Another example: HIV virus is very complex compare to some other viruses. But because of
the complexity it evades immune response. This is why it is so successful.
For  humans  there was some discussion that hernias are direct
problem of  erect posture. So erect posture leads to disadvantage. From the other hand erect
posture  frees the hand, that allows to produce the tools and this gives huge advantage in the
survival in natural world by creating possibility to adapt to virtually any condition.

All high animals have  just 4 extremities. Obviously it is enough in cutthroat environment. Growing
additional couple of extremities  will consume much more resources and take much more time, not
giving big advantage  in speed or protection of an animal.
Thus, danger of losing one of the limb is not so big as the danger to not proliferate fast enough.

Bilateral design of majority of animals was chosen at the level of primitive species. Though lower
multicellular  organisms may have 3-side symmetry or 5-side symmetry like sea stars. Often
something chosen in the selection becomes building blocks for higher species.
Maybe  5-side symmetrical tiger would be possible but it requires  intermediate chain.

The same is true for the mechanism of Death. Once chosen,  it is replicated in all  higher organisms.
It should give an advantage in ultimate species survival. Otherwise it loses sense and species would
be wiped out.

So, what is advantage for Death?

Immortal and childless animals are growing and growing.   Mutations  lead to species producing
offsprings. They have disadvantage of been more complex. But advantage of  proliferating animals is
occupation of vaster areas. They displace immortal and childless species.
Now, if  proliferating animals are immortal they do not give offsprings enough space or food.
Next steps of mutation brought mechanism of programmed death at certain age.

Let consider situation when out of  two species one is immortal and producing offsprings, other is
mortal with offspring production as well.
Mortality looks like an obvious disadvantage in stable environment. Adult organism are more
powerful and more protected compare to juvenile.

However, next catastrophe will wipe out all the species altogether. Only some mutants can adopt to
new
environment.
But "mutant" for a multicellular organism imply that the mutation happened when the organism had
only one cell (the stage of fertilized oocyte for example). Otherwise it needs precisely the same
mutations on the level of several cells - infinitely impossible event.

Thus, any species that had the mechanism to allow next generation to flourish (mechanism of
programmed Death) will have advantage. They produce more offspring than immortal species.
Death of parents spares  the space and  nutrition. Immortals can die form accidents only. It is too
long for children to expect a vacancy.

Then, having more offspring automatically means having more mutations (by mere number of
events). Having more mutations means more probability for adaptation to the new environment.

Immortal multicellular organisms, even if they existed in the past, were quickly wiped out by
accidents and overgrown by mortal multicellular organisms. 

Again all this is just a hypothesis.




Aging


How  did aging appeared?

If mechanism of Death was chosen by evolution, how was aging chosen?

Why for example at certain age animals do not die  instantly and just give  the room for the next
generation.
Nurturing is one explanation at least for human society.

But lets say new generation of young adults already reached puberty . Why would not  old
generation die in
an instant. Why do they age?

Actually there is nothing impossible. Some  butterflies grow couple years,  mate,
produce new generation and die in one day.
Some plants  live 1 year, some live 2 years and then invariably die. Some live 1000 years.
Lemmings, having  overgrowth of population, just  go and
massively commit suicide in the sea. No questioned asked.


Why aging in other animals take several years?

It might be selected by cut-throat evolution during another catastrophe.

A relatively mild Cataclysm happened in the same time when all adults died programmed to do so.
Older animals would survive the disaster. But they died instantly from internal timer. Younger
generation did not tolerate the mishap.

All species that did it - just disappeared. Younger animals were not adapted  to new
conditions.
Only animals with mechanism of aging  were selected. Maybe the aged animals are not so good
as younger ones. They already slightly deteriorated. But it is  better than nothing.

Anyway. It  makes sense why  Death and Aging  were selected during the Evolution.



Biological clock


Obviously you need biological clock to time  and alarm the point when you need to die and when
you need to age.
Aging  is the same death as  in those one-day butterflies, but  a little bit prolonged one.   Not
immediate kill but gradual shut down  of protection mechanisms - immunological, genetical repair,
and just other  necessary functions of proliferation. As soon  as those functions turned off -
an organism starts  to deteriorate and wear off.

Interesting that our organism is not a static thing. We probably replace all  molecules in our body
within relatively short period of time. Obviously water is replaced completely every couple months.
Water is 60% of our body. Other molecules are replaced as well.
Thus,  human being is just some unstable chemical organisation around genetical information.
Molecules swirl and disappear like crazy.
DNA in cells maybe is not changed so fast, but the cells themselves change  quickly in most of
cases. There is no need to age just because of wearing.

Biological clocks are necessary and there is no doubt they exist.

The theories of aging  that consider death as a sequence of  events caused by external causes -
like oxidations  and  accumulation of genetical errors should anyway include  the idea of
Internal Biological Clock as well.
Simple  example  is  menopause in women.  Somewhere at 40-55 all women undergo menopause.
Unavoidable. Though there were reports of childbirth at 60 yo but considering  one chance in
billions, it is just  a joke.
Males have their equivalents too.
All people after certain age change their immunological, hormonal, metabolic levels and so on.
Majority of organs decrease their function . Thymus involute. Fat cells overgrow others,  muscle
cells degenerate etc.
The same happens to all other multicellular animals and plants.

Now the real question:  Is it possible to break the  biological clock or at least slow
down and rejuvenate?





Possible methods of rejuvenation.


1. Telomeres lengthening and other genes regulation.

Several genes that might play role in aging described recently. Telomerase encoding genes are the
most discussed. In theory, if we lengthen the telomeres (these are the ends of chromosomes that are
shortened with each division by very interesting mechanism), we could overcome cell tendency to
lose ability for proliferation. Hence, we have "immortal cells". Hence, immortality or at least
longevity is reached - cells divide indefinitely and repair organism. Basically,  to restore telomeres
length we need to activate production of or to introduce into the cell an  enzyme named telomerase.
To reach each cell in the body we would use viral vectors or smallRNA.
Working with virus production and then cell transduction I would say it is difficult, very difficult.
There are too many obstacles to use them efficiently. SmallRNAs would be more promising. In any
case, when you go from  theory to practice problems rise exponentially.
Biggest question:  will it work? Immortal cell (cell which is able to divide indefinitely) does not mean
immortal body. One of the outcomes would be uncontrollable cell growth and, hence, cancer.
Actually scenists  discuss telomeres mostly in connection with cancer. They are very cautious saying
about the rejuvenation. Mass media extrapolates  the findings  to
possibility of longevity and further  rejuvenation.

My guess would be that telomeres are some of the "final cogwheels" in the Internal biological clock.
It is useful to divide the clock on intracellular and whole body levels just for understanding. But in
reality the levels are not separable, they continue one into another. It is easy to understand that
intracellular production of any protein, any enzyme depends on transmembrane regulation.
Hormones activate receptors that activate DNA transcription and protein production.
Other genes that play role in cellular aging are described recently as well.
I do not know how to use this method at present time

2. Hormone replacement.

When I studied in med school, several time I met mentions about hormonal side effects that cause a
person to look much younger.
Well known for example are:
If a woman in menopause suddenly resumes menstrual cycles and looks younger than her age, we
might suspect an ovarian Tumour producing excess of estrogen.
If a person has thyrotoxicosis - excess of hormone thyroxine - he/she loses weight and looks much
younger
Persons with manic phases of bipolar disorder look younger, more energetic.
Certain hormones - corticosteroids for example - might be increased or decreased  in elderly.
Couples that look younger were shown to have increased level of sexual relationships. Again, that
supposedly should significantly change the hormonal levels.

If we give a person some of those synthetic hormones -  estrogen, thyroxine - we can sometime
reproduce the younger looks.

It is not the same as plastic surgery or skin creams and massages. It invigorates whole body. Skin
as well.
There is heavy price - recent debates are waxing and waning - if estrogen replacement increases or
decreases risks of cancers (uterine, breast, ovarian etc.) Thyroxine excess takes heavy tall on
cardiovascular system.
Psychiatric disorders (mania for example) was linked to ceratin changes in neurotransmitter levels.
Certain psycho tropic drugs change the ratios. Supposedly they can be employed for rejuvenation.
Unfortunately, you would rather receive a psychosis than younger look.

On internet you can find massive information about hormones that rejuvenate you.
I saw some websites that list practically any major hormone:
estrogen
androgens
corticosteroids
growth hormone
thyroxine
DHEA
melatonin
and some others as a drug that leads to rejuvenation.
The websites often have references to scientific literature.
Most of the websites just sell you those hormones or offer courses of hormone therapy.

My personal opinion would be that blind replacement can cause more harm than  lead to any
desired rejuvenation.

Useful information that I would extract from the item is that A) Practically every hormone in the
body is involved.
B) Certain means of rejuvenation are actually documented. That gives hope.

I do not use hormones personally. If I would employ them, I would use them in very small quantities
to mimic physiology and in special controlled conditions that I will describe below.
The doses that are used in regular therapy are actually way to high compare to the natural levels of a
healthy organism.

3. Antioxidant therapy.

As we discussed, in the best case antioxidants are the method for longevity, not rejuvenation.
I use mega-doses of antioxidants available over the counter. I use only those  that I consider safe. I
have a lot of doubts they give measurable effect.
Anyway I use them. Would not throw away.

4. Caloric restriction.

I use it as well. 60% of regular meal - is way to much - you would barely move your feet. I regulate
it.
My opinion: if you increase your activity, you can eat more. Just balance your food and motions.
Though I might be  wrong.


5. Hypothalamic regulation.


This is scary level. Hypothalamus is the area of your brain that sits on the base of your skull.
Why do we discuss hormones above? Well, hypothalamus contains hundred of centres that regulate
your metabolism and first of all, hormonal status.
Studies of hypothalamus are complicated - mostly there are postmortem studies of brain sections.
To perform functional study you need to use micro-neurosurgical techniques (still very rude) and try
to register changes in a hundred or so cells out of several billions. Changes would include minuscule
production (nano and Pico grams) of poorly defined neurotransmitters. The infinitely small quantities
of those neurotransmitters are produced by only few out of hundreds cell connections and exist only
several nanoseconds.
In general to study this area is more difficult than to look for a needle in a haystack. If you consider
that there are only few centres have appropriate expertise, you would understand why the progress
is relatively slow. Even fewer labs study rejuvenation.
Though there were reports that transplantation of hypothalamic tissue from younger animals to older
could result in rejuvenation-like effect.

I mention this level because A) hypothalamus regulates production of bunch of intermediate
messengers that in turn activate hypophysis, that activate adrenal glands, ovaries, thyroid gland etc
(maybe many other areas in th body) that in turn produce the hormones discussed above. (Full
description you could find in a textbook of Endocrinology)
B) because of complexity of hypothalamus it is a good candidate were a "supreme cogwheel" of
biological clock may reside. One hint would be proximity of epiphysis - small pea-like gland that
produce melatonin and regulates circadian rhythms. It would not be surprise that all that is
interconnected in the hypothalamus.
One of objection  - what is  about worms? They do not have hypothalamus as it is. But they age
and die as well as humans. Explanation would be that worms also have nervous system and some
other part of it may play the central role in the biological clocks. Another explanation would be that
there is no such centre at all, everything happens on the lower levels.

I was interested in methods of rejuvenation since childhood. Maybe I read too many science fiction.
When I was in medschool a saw some mentions of unusual phenomenon in magnetic radiation.
To make story short, eventually I found a book  - 400 pages of fine script.
I found the book in Central Medical Library in Moscow.
Even our relatively large library in Russian State Med U did not have this book.
In addition I read the dissertation thesis, that were related to that book.
Several Russian scientists developed relatively complicated theory, done many hundreds of
experiments.
Theory is a little bit complex, involves discussion of stress and adaptation syndrome. Obviously
hypothalamic area also discussed.
I am not big fan of that theory. But result the authors described were striking:
they were able to restore menopause in  rats, prolong rats life span almost twice and more
important repeat it again and again. By the description from the book "rejuvenated rats" looked the
same as young, but were just bigger in size.
Well, you saw a lab rat, you know that older rats have rare hair, bleak eyes, are in menopause etc.
Young rat, in opposite, would have bright red eyes, very sleek hair, menstruations, etc.
This is no surprise - all growth of hair, production of oil by skin, production of moisture in lacrimal
glands and so on are regulated by the  hormones. The same is true for rats as well as for humans.
Authors described the methods that they found mostly empirically.
Moreover they tested some methods  on humans and described treatment of variety of diseases -
asthma, gastritis, peptic ulcers, low grade skin cancers (non-melanoma) just to name few.
As I understand they did not describe rejuvenation in humans. Though later there were some
mentions in  literature. Authors describe zones of activation, training and stress.
In general the methods are difficult to apply - they require permanent control of organism condition
for many months, maybe years. Control of hormonal level and control of blood work. In addition
parameter  would vary from individual to individual. In modern medicine it is much faster and more
reliable to cut out skin cancer with knife or give powerful proton pump inhibitors for a  gastric ulcer.

I realized that I used some of the methods described, even before I read that book. Since then I
added some more methods for my own use. Mostly I use the methods that I feel safe and efficient.
For example transplanting of hypothalamus would  be out of question. It is a wacking unsafe
procedure.
Recently there were reports that DHEA, melatonin or other hormones could make miracles: older
people feel as young.
I would wait confirmation of the effects. Some authors suggest  rejuvenation may be
reached by individually designed cocktails of hormones.
I would think that any hormone given makes more mess than regulation. Practically every hormone
described have loops and pathways that prevent overproduction. If you just replace the hormones
by
giving the substitute, you wack the production of your own hormones those that injected as well as
many others. All hormones are complexly interrelated.
Individual cocktails would be better, but  the doses described in literature are wacking.
Again I might be wrong. Maybe everything is so simple - just get cocktail and you will jump as
a newborn bunny.

Considering different information from different sources, I would propose following theory:

To rejuvenate yourself you need to recreate the hormonal and metabolic level  of young organism.

This is actually very difficult to reach: your organism have all the means to produce all the necessary
hormones. Levels of hormones in elderly are not so far from the levels in youth actually. The
difference is several percentages, not several folds.  But biological clock switch to underproduction.
The idea is to cheat the biological clock and to restore the youth levels.
The most difficult part is to get into the window of optimal production.
You can not go too low - just wouldn't have any effect on rejuvenation. You can not go above the
optimal level - it will cause stress and even more deterioration.
So approach is very individual. There is no quick effect - you can not rejuvenate from looking 50 to
looking 30 overnight. You really need a lot of commitment and time.
The methods which you choose are not of so great importance. Practically any action toward your
body would cause changes in your metabolic and hormonal status. Physical exercise, herbs, yoga,
hormones (low doses) etc - we could list thousands.

Personally I used available antioxidants, slight caloric restrictions, herbs (some of substances in
herbs like ginseng, Echinacea, etc mimics hormones in their chemical structure), self-hypnosis,
neurolinguistic programming (NLP),  auto-training, different exercises, lactic bacteria, etc. Important
is the ability to adjust the level of treatment - reduce the dose of herbs or increase dose of exercise
etc.

Methods of control  are of much great importance.
The described methods include measurement of hormonal level and of certain fractions of cells in
the blood work(that is also regulated by hormonal and metabolic reactions)

If  I would have a clinic of rejuvenation therapy I would probably use many of those methods - they
are objective and reproducible. But they require blood work.

For individual use there some subjective methods exist. They are described in literature as:
If you are in the condition of rejuvenation you feel great energy, hunger, restlessness. Your sleep
would be normalised, you are fighting regular infections much more efficiently - inflammatory
reaction might be exaggerated, but they last shorter time. Your eyes would glisten as in youth. You
will feel constant euphoria and need in doing something.
There are some other  subjective criteria.
Personally, I would add: you will notice that your nails and hair grow like crazy in those conditions.

The biggest problem is how to hold in this condition of rejuvenation for a prolonged period of time.
You really can not go overboard - super activation would just wack all your system. You can not
go under activation - just no any useful effect. You need to get it is just right.
Solution is tight, continuos control.
Boy, it's really tough in the modern stressful environment (would not describe all the junk that I had
in recent years), but it is possible. Dedication is your solution.
Some formulas to calculate doses of stimuli are described in the literature, but I feel it is easier to go
with individual subjective feelings.
Below there is an animation that contains some simple elements of NLP and might help to tune a
brain a little.


Now, what did I get by using those methods?
Well not much, but I hope for more.
At present time I am 35 yo I do not really have good control group in my personal experiment.
Sometime people asked me if I am younger than my brother (He is younger than me for several
years). Maybe it is some sort of control group.
When I come into a bar (can not say often, maybe once in a year with friends) waiter often  asks
my ID. It is funny also because I do not use alcohol. So, to drink a Coke I need to show ID, being
35 yo.
Sometime people ask me are you 18 or 19
When I played water polo at Yale, undergrads told me they thought I was freshman (as I guess also
18 or 19). That time I was 33. Teenagers often start to talk to me in the way if I am another
teenager.
And so on.

So, in general it is really hard to measure rejuvenation. All measurements are mostly subjective.
Even
in  well designed scientific trial you find either some description of biochemical changes (which
doesn't say you anything about rejuvenation as it is) or some subjective descriptions  - "patients felt
much better, felt rejuvenated and invigorate". How much rejuvenated? How many years were
reversed? It is very difficult to estimate.
If I say to a person that I  have MD and PhD, he says "aha". Mentally they immediately adjust and
now they say that I am probably 30. OK. Sometime they just say: you look so young.

Again 35 is not the age where you really can brag that the methods worked. I would wait couple
more decades to say that there was measurable effect. Unfortunately. You do not have the right for
a
mistake. If a method did not work , you can not go back and change it. You already are behind for
several years.

But my personal, subjective feeling is that the methods do work. I believe that it is possible by tight
control maintain the level of younger organism. It  just requires a lot of work. 

Below there is animation that might help to tune in the desired "optimal rejuvenation condition".
Animation requires Flash file player
I use that animation for myself.
I doubt that it will necessarily help you. Maybe 1 out of 10. If you are not sure, don't' touch it.
The animation contains some simple NLP information.
Animation
Basically looking at the animation couple times a week for 5 min should start the tuning process (at
least I feel so).
You should target at the condition when you feel inexhaustible energy, euphoria, normalized sleep,
and some feelings that is hard to describe - like a fine tingling in your fingertips, subjectively it feels
like a pouring energy. It is difficult to explain, but when you feel you understand it.
If you do not feel anything, it means it did not work, forget it.

So to finish I would say that Rejuvenation is difficult but probably possible to reach  - at least in a
certain degree.    A.K.2004 copyright
Animation
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