[extropy-chat] Re:Resuscitation: and Armchair Cryonicists-hypothermic liferaft

Extropian Agroforestry Ventures Inc. megao at sasktel.net
Wed Dec 15 13:56:05 UTC 2004


United States Patent Application 	20040097534
Kind Code 	A1
Choi, Byung-Kil ;   et al. 	May 20, 2004

------------------------------------------------------------------------
Composition for the protection and regeneration of nerve cells 
containing berberine derivatives

Abstract

Disclosed is a composition for protecting nerve cells, promoting nerve 
cell growth and regenerating nerve cells comprising berberine, 
derivatives thereof or pharmaceutically acceptable salts thereof. The 
composition has protective effects against apoptosis of neuronal stem 
cells and differentiated neuronal stem cells, an effect of inducing the 
regeneration of nerve cells, a regenerative effect on neurites, a 
neuroregenerative effect on central nerves and peripheral nerves, a 
reformation effect on neuromuscular junctions, and a protective effect 
against apoptosis of nerve cells and a neuroregenerative effect in 
animals suffering from dementia and brain ischemia. Therefore, the 
composition can be used as a therapeutic agent for the prevention and 
treatment of neurodegenerative diseases, ischemic nervous diseases or 
nerve injuries, and for the improvement of learning capability.

&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&

[0150] Next, the head of the rat was fixed on a stereotaxic apparatus to 
operate on the occiput, and then the tail was fixed so that it descended 
downwardly at an angle of 30.degree.. After incising the occipital bone, 
an electrocauterizing needle having a diameter of 1 mm or less was 
inserted into the alar foramina positioned at lower part of the first 
cervical vertebra under the occipital bone. At this time, this approach 
must be carefully done so as not to damage the muscles in the alar 
foramina. Thereafter, the vertebral artery was electrically cauterized 
by intermittently applying current. After the complete 
electrocauterization of the vertebral artery was confirmed, suturing was 
carried out using operating clips. After 24 hours, the operating clips 
were removed. Finally, the common carotid arteries were occluded using 
the silicone tube rings for 10 minutes to induce ischemia. If light 
reflex did not disappear within 1 minute, the cervical portion was 
further tightly sutured. Rats which did not show the complete 
disappearance of light reflex were excluded from the experiment because 
they underwent no damage to the CA1 region. After 10 minutes, the common 
carotid arteries were loosened to reperfuse. For 20 minutes after the 
reperfusion, loss of consciousness was observed. At this time, only rats 
which showed consciousness loss period within 20.+-.5 minutes were 
selected for subsequent experiments.

&&&&&&&&&&&&&&&&&&&&&&&&&&&&&

2) Experimental Results

(1) Concentration of Berberine, Influence of Body Temperature and 
Ischemia Inducing Time

[0157] The highest concentration of berberine was set to 300 .mu.g/0.1 
kg, and 600 .mu.l (1 mg/ml) of berberine was intraperitoneally injected 
to white rats weighing 200 g. In order to determine an optimal ischemia 
induction time, 2.about.3 rats were selected and ischemia-induced over 
5, 10, 20 and 30 minutes, respectively. 1 week after reperfusion, they 
were sacrificed and their hippocampal tissue sections were obtained to 
observe the number of damaged nerve cells. 10 minutes after ischemia 
induction, damaged pyramidal cells in the hippocampal CA1 region were 
found to be reduced to 1/4 of their original numbers. The ischemia 
induction time of 10 minutes was determined to be most optimal for 
evaluating the effects of berberine.

[0158] For statistically analyzing the effects of berberine, a sham 
operated group having undergone an operation in the same manner without 
ischemia induction was used. For comparing the effects of berberine, a 
control group administered with physiological saline at the same dose as 
berberine was used. Berberine was intraperitoneally injected into all 
experimental groups.

[0159] It is well known that reduction in body temperature during 
ischemia induction prevents damage to nerve cells in the hippocampus and 
thus exhibits neuroprotective effects. Therefore, in order to evaluate 
the neuroprotective effect of berberine, after ischemia induction and 
reperfusion, the body temperature of all rats was maintained at a 
constant (37.+-.1.degree. C.) for 6 hours.

(2) Observation of Damaged Nerve Cells

[0160] When ischemia was induced by 4-VO and then reperfusion was 
performed, nerve cells in the neocortex, striatum, hippocampal CA1 
region and cerebellum were damaged. Among them, pyramidal nerve cells in 
the hippocampal CA1 region were the most susceptible to the induced 
ischemia, and started to undergo cell death 72 hours after reperfusion. 
In order to observe delayed neuronal death in the hippocampal CA1 
region, 1 week after reperfusion, the time when almost all nerve cells 
were damaged, white rats were sacrificed and tissue sections from the 
hippocampus were observed under an optical microscope. In a sham 
operated group having undergone no ischemia, normal hippocampal nerve 
cells were observed in the stratum pyramidale (490 .mu.m long)(see,A and 
B of FIG. 21).

[0161] C and D of FIG. 21 as control groups show apoptosis. When cells 
are induced to undergo apoptosis by an external or an internal stimulus, 
they shrink to lose their original shapes. This shrinkage breaks the 
junctions with other adjacent cells so that the interaction between 
cells is disrupted. When the shrinkage proceeds to some extent, the cell 
membranes form apoptotic bodies like a bulla. In the hippocampal CA1 
region of the control group administered with physiological saline (D of 
FIG. 21), it was observed that nerve cells underwent apoptotic 
morphological changes after ischemia induction. In addition, it was 
observed that tissues was relaxed and separated from adjacent cells, 
unlike B of FIG. 21. From these observations, it was confirmed that the 
cell bodies of nerve cells lost their original pyramidal shape and were 
condensed, thereby appearing to be single cells. Furthermore, it was 
confirmed that subsequent nuclear chromatin condensation and nuclear 
envelope collapse led to apoptosis of nerve cells. On the contrary, 
nerve cells in the hippocampal CA1 region administered with berberine 
were similar to normal cells in terms of their morphology (see, E and F 
of FIG. 21). At this time, because necrotic nerve cells around the CA1 
region were very difficult to distinguish from microglias, only viable 
pyramidal nerve cells in the CA1 region were counted. In F of FIG. 21, 
separated cells were observed above and below the hippocampal region and 
cell bodies were condensed. This demonstrates that the damage to nerve 
cells was great enough to induce apoptosis. Nevertheless, it was 
observed that a great number of nerve cells were protected from 
apoptosis and their original pyramidal morphology was maintained. This 
suggests that berberine has a protective effect against damages to nerve 
cells in the hippocampal CA1 region induced by 4-VO. Although it was not 
confirmed what stage during apoptosis influences nerve cell survival, it 
was certain that berberine has a significant protective effect against 
apoptosis of nerve cells (see, E and F of FIG. 21).

(3) Protective Effect of Berberine Against Damage to Nerve Cells

[0162] In order to examine the neuroprotective effect of berberine after 
ischemia induction, berberine was intraperitoneally injected 0 and 90 
minutes after ischemia induction.

[0163] In the sham groups, the density of viable cells was measured to 
be 308.+-.6.6 cells/mm.sup.2 (at 37.degree. C.). In the control groups 
administered with physiological saline, the density of viable cells was 
measured to be 28.+-.3.8 cells/mm.sup.2 (at 37.degree. C.). There was 
cell loss in these two groups. On the other hand, in the experimental 
groups administered with berberine, the density of viable cells was 
measured to be 257.+-.9.6 cell/mm.sup.2. In conclusion, berberine was 
determined to have a significant neuroprotective effect (p<0.05).

[0164] As described above, the composition according to the present 
invention regenerates axons and dendrites of nerve cells, thereby having 
a protective effect against nerve cell injuries, a positive effect on 
nerve cell growth and a regenerative effect on nerve cells. In addition, 
the composition according to the present invention can be used as a 
therapeutic agent for the prevention and treatment of neurodegenerative 
diseases or nerve injuries, in particular, dementia, Parkinson's 
disease, Alzheimer's disease, epilepsy, palsy, ischemic brain diseases, 
trauma to the spinal cord and peripheral nerve injuries.

[0165] Although the preferred embodiments of the present invention have 
been disclosed for illustrative purposes, those skilled in the art will 
appreciate that various modifications, additions and substitutions are 
possible, without departing from the scope and spirit of the invention 
as disclosed in the accompanying claims.

* * * * *

Aside from its carrier capacity DMSO was chosen for its ability to 
inhibit damage from a light freeze

There are a number of complementary chemistries besides from the ones 
cited readily available.



Eugen Leitl wrote:

>On Wed, Dec 15, 2004 at 03:01:49AM -0600, Extropian Agroforestry Ventures Inc. wrote:
>
>  
>
>>The concept goes thusly:
>>The liferaft =
>>-sleeping /body bag like sack made so that no 2 arms or legs touch each 
>>other or body
>>-zipper + ziplock seal
>>- control/RFID biomonitor keypad with on outside
>>-stage 1- evacuate liner to ensure good skin contact with sack
>>-person put inside without outer clothes , shoes etc
>>-put cooling hood or cap over all of head less face, face cover ziplock 
>>cover after body cooled off
>>-start in 2 parts; activation of emergency cooling packs layer of sack 
>>to quick cool body; hood cooling cycle
>>    
>>
>
>Useless. This gives you no advatage over an ice bath.
>
>  
>
>>co2 based for higher cooling rate
>>    
>>
>
>More than useless. You can't go below 0 C, or you'll get freezing injury.
>
>  
>
>>-infusion of adjuvants may include:
>>Caffeinol as neuroprotectant; berberine in DMSO solution as 
>>neuroprotectant; cannabidiol in DMSO solution as neuroprotectant
>>-optional defib cycle to pump neuroprotectants into cooling  body uniformly
>>    
>>
>
>You have to maintain the circulation. Best do achieve this is life support.
> 
>  
>
>>The adjuvants are designed to allow the brain to survive a longer 
>>cool-off time than the usual 3-5 minutes.
>>    
>>
>
>Sorry, but your science is garbage. I'm being delibertely harsh here, because
>otherwise you won't get the message.
>
>  
>
>>as well as allow for easier re-start of body by hospital medical team
>>-once body temp is near 32F optional external hookups to maintain cooled 
>>body during extended transport
>>
>>Once the working prototype is designed and tested  , the actual mfg 
>>costs may be quite reasonable
>>    
>>
>
>If you have to live in the sticks, you have to rely on people. No machinery
>is going to help.
> 
>  
>
>>Morris Johnson
>>
>>Hara Ra wrote:
>>    
>>
>>>For Eugen:
>>>
>>>   Thanks for taking my point and clarifying it. If you read my sig 
>>>file, I don't think I am an "Armchair cryonicist". Please correct me 
>>>if you think so, and explain why.
>>>      
>>>
>
>Of course I wasn't commenting on what you wrote, but on periodical resurgence
>of well-meaning-but-clueless armchair cryonicists.
>
>  
>
>>>PS We currently do no longer use the Thumper, which is an obscene 
>>>bastard of equipment fully capable of major injury to both patient and 
>>>rescue team. Respiratory support is no longer in the protocol, because 
>>>      
>>>
>
>Oh yeah, if the cup breaks off you'll get a massive metal rod puncturing the
>ribcage.
>
>  
>
>>>basically all patients are over the 4 minute limit, and restoring O2 
>>>is a bad idea. We use an Ambi product, a suction cup with handles, to 
>>>      
>>>
>
>Not if you add neuroprotectants via IV push, and maintain artificial
>circulation.
>
>  
>
>>>maintain circulation for the 3-5 minutes it takes to circulate the 
>>>medicines (a proprietary cocktail of anticoaguants, clot busters and 
>>>other stuff)
>>>      
>>>
>
>
>  
>
>------------------------------------------------------------------------
>
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>extropy-chat at lists.extropy.org
>http://lists.extropy.org/mailman/listinfo/extropy-chat
>  
>

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