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<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: #548dd4; mso-themecolor: text2; mso-themetint: 153">-----Original
Message----- <BR>From: Alejandro Dubrovsky <BR>Sent: Friday, September 6, 2013
9:53 AM <BR>To: cryo@postbiota.org <BR>Subject: [cryo] Fwd: [ExI] Steve Van
Sickle's presentation at SENS6 <BR><BR>-------- Original Message
--------<BR>Subject: [ExI] Steve Van Sickle's presentation at SENS6<BR>Date:
Fri, 6 Sep 2013 01:40:34 -0400<BR>From: Rafal Smigrodzki
<rafal.smigrodzki@gmail.com><BR>Reply-To: rafal@smigrodzki.org, ExI chat
list<BR><extropy-chat@lists.extropy.org><BR>To: ExI chat list
<extropy-chat@lists.extropy.org><BR><BR>Yesterday I had the pleasure of
attending Steve Van Sickle's<BR>presentation at SENS6, reporting his results
with cryonic preservation<BR>of kidneys. Steve developed a technique for
persufflation of organs<BR>perfused with cryoprotectants using cold helium. He
was able to cool<BR>pig kidneys to liquid nitrogen temperature, achieving
total<BR>vitrification while *avoiding* fractures! I was absolutely floored
by<BR>his work, easily the most important presentation at SENS6 so far.<BR>Steve
also described calculations for hyperbaric persufflation, which<BR>might allow
vitrifying a human body in *a few minutes*!</SPAN><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: #548dd4; mso-themecolor: text2; mso-themetint: 153; mso-fareast-font-family: 'Times New Roman'"><o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: #548dd4; mso-themecolor: text2; mso-themetint: 153; mso-fareast-font-family: 'Times New Roman'"><o:p> </o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'">There's
only one small problem (at least for me) with this "remarkable breakthrough", or
more particularly, with the crediting of it.<o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'">When
Steve Van Sickle was President of Alcor, and Tanya Jones was in charge of acute
patient care, I engaged in an extended and frustrating series of emails with Van
Sickle, copied to Tanya and to Alcor Director Dr. Brian Wowk. The subject of
this correspondence was my urging Alcor to switch to using gas perfusion
following cryoprotective perfusion of patients, in order to achieve both more
uniform cooling, as well as much faster cooling and very possibly the reduction
or elimination of fracturing injury in patients.<o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'">The
reasons I suggested this were not merely theoretical. When I was working as a
researcher for Manrise Corporation (Fred and Linda Chamberlain) in the
mid-1970s I had conducted gas perfusion experiments on rabbit kidneys and heads
following the work of Schimmel, et al. with gas perfusion of rat kidneys; <A href="http://www.ncbi.nlm.nih.gov/pubmed/5872343"><SPAN style="COLOR: blue">http://www.ncbi.nlm.nih.gov/pubmed/5872343</SPAN></A> ,
that of of Hamilton and Lehr with gas peerfusion of canine small bowel <A href="http://www.ncbi.nlm.nih.gov/pubmed/4702169"><SPAN style="COLOR: blue">http://www.ncbi.nlm.nih.gov/pubmed/4702169</SPAN></A> and
of the work of Guttmann, et al., using hedlium perfusion in canine kidneys: <A href="http://www.ncbi.nlm.nih.gov/pubmed/1259562"><SPAN style="COLOR: blue">http://www.ncbi.nlm.nih.gov/pubmed/1259562</SPAN></A> and
<A href="http://jama.jamanetwork.com/article.aspx?articleid=661256"><SPAN style="COLOR: blue">http://jama.jamanetwork.com/article.aspx?articleid=661256</SPAN></A> .<o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'">This
work demonstrated that uniform cooling rates on the order of 2-3 deg C min were
achievable and, as I noted in my correspondence with van Sickle at that time,
there was no evidence of fracturing, even in kidneys loaded with 25% w/v
cryoprotectant.<o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'">I
would also point out my extensive discussion of the idea of gas perfusion in
cryonics in this post on Cryonet during the "cooling fluids debate" that took
place there on 12/17/92:<o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"><A href="http://www.cryonet.org/cgi-bin/dsp.cgi?msg=1465"><SPAN style="COLOR: blue">http://www.cryonet.org/cgi-bin/dsp.cgi?msg=1465</SPAN></A><o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'">I
pointed out that this technique was far easier to implement than was
perflurochemical (PFC) perfusion and that it had the advantage of having already
been proven out from a practical standpoint not only by my work in the
mid-1970s, but by work done by Pegg, et al., in the in 1978; <A href="http://www.sciencedirect.com/science/article/pii/001122407890086X"><SPAN style="COLOR: blue">http://www.sciencedirect.com/science/article/pii/001122407890086X</SPAN></A> .
What's more, experiments conducted in recent years using oxygen persufflation of
hypothermically (non-frozen) stored organs had demonstrated that putative
problems with gas emboli, or endothelial cell dehydration, were, in fact NOT
problems and that organs perfused with gas could be transplanted with the
long term support and survival of the animals being a uniform
outcome:<o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"><BR>1:
Srinivasan PK, Yagi S, Doorschodt B, Nagai K, Afify M, Uemoto S, Tolba
R.<BR>Impact of venous systemic oxygen persufflation supplemented with nitric
oxide gas<BR>on cold-stored, warm ischemia-damaged experimental liver grafts.
Liver Transpl.<BR>2012 Feb;18(2):219-25. doi: 10.1002/lt.22442. PubMed PMID:
21987402.<o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"><BR>2:
Minor T, Akbar S, Tolba R, Dombrowski F. Cold preservation of fatty
liver<BR>grafts: prevention of functional and ultrastructural impairments by
venous oxygen<BR>persufflation. J Hepatol. 2000 Jan;32(1):105-11. PubMed PMID:
10673074.<o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"><BR>3:
Stegemann J, Hirner A, Rauen U, Minor T. Gaseous oxygen persufflation
or<BR>oxygenated machine perfusion with Custodiol-N for long-term preservation
of<BR>ischemic rat livers? Cryobiology. 2009 Feb;58(1):45-51.
doi:<BR>10.1016/j.cryobiol.2008.10.127. Epub 2008 Oct 17. PubMed PMID:
18977213.<o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"><BR>4:
Minor T, Olschewski P, Tolba RH, Akbar S, Kocálková M, Dombrowski F.
Liver<BR>preservation with HTK: salutary effect of hypothermic aerobiosis by
either<BR>gaseous oxygen or machine perfusion. Clin Transplant. 2002
Jun;16(3):206-11.<BR>PubMed PMID: 12010145.<o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"><BR>5:
Lauschke H, Olschewski P, Tolba R, Schulz S, Minor T. Oxygenated
machine<BR>perfusion mitigates surface antigen expression and improves
preservation of<BR>predamaged donor livers. Cryobiology. 2003 Feb;46(1):53-60.
PubMed PMID:<BR>12623028.<o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"><BR>6:
Gong J, Lao XJ, Zhang SJ, Chen S. Protective effects of L-arginine
against<BR>ischemia-reperfusion injury in non-heart beating rat liver graft.
Hepatobiliary<BR>Pancreat Dis Int. 2008 Oct;7(5):481-4. PubMed PMID:
18842493.<o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"><BR>7:
Nagai K, Yagi S, Afify M, Bleilevens C, Uemoto S, Tolba RH. Impact
of<BR>venous-systemic oxygen persufflation with nitric oxide gas on steatotic
grafts<BR>after partial orthotopic liver transplantation in rats.
Transplantation. 2013 Jan<BR>15;95(1):78-84. doi: 10.1097/TP.0b013e318277e2d1.
PubMed PMID: 23263502.<o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"><BR>8:
Minor T, Isselhard W, Klauke H. Reduction in nonparenchymal cell injury
and<BR>vascular endothelial dysfunction after cold preservation of the liver by
gaseous <BR>oxygen. Transpl Int. 1996;9 Suppl 1:S425-8. PubMed PMID:
8959878.<o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"><BR>9:
Saad S, Minor T, Nagelschmidt M, Fu ZX, Kötting I, Paul A, Troidl H,
Isselhard<BR>W. [Revitalizing donor livers after cardiovascular arrest with
venous oxygen<BR>persufflation]. Langenbecks Arch Chir Suppl Kongressbd.
1998;115(Suppl I):705-8. <BR>German. PubMed PMID:
14518345.<o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"><BR>10:
Tolba RH, Schildberg FA, Schnurr C, Glatzel U, Decker D, Minor T.
Reduced<BR>liver apoptosis after venous systemic oxygen persufflation in
non-heart-beating<BR>donors. J Invest Surg. 2006 Jul-Aug;19(4):219-27. PubMed
PMID: 16835136.<o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"><BR>11:
Sun HW, Shen F, Zhou YM. Influence of perfusion by gaseous
oxygen<BR>persufflation on rat donor liver. Hepatobiliary Pancreat Dis Int.
2006<BR>May;5(2):195-8. PubMed PMID: 16698574.<o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> Van
Sickle's response was, essentially, to argue that gas perfusion simply could not
compete with the exchange capability of a liquid - even a liquid with relatively
poor heat carrying capacity, such as a perflurochemical (PFC), or a mixture of
PFCs. These remarks were presumably made on the basis of a patent by Brian Wowk
and demonstrating the utility of PFC for improving heat exchange during
cooling of organs and whole animals. <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'">My
efforts to point out to Steve that this technique was severely limited
because:<o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'">*
Because of the increasing viscosity, and ultimately the solidification of the
PFCs at temperatures right around the critical glass transition point for M-22,
the ability to remove heat from the patient is lost at precisely the point
during cooling when it was most needed.<o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'">*
Perfusion of PFCs was incredibly uneven in the tissues due to a short circuiting
effect that occurred when very low viscosity PFC first opened a channel of flow
between the arterial and venous circulations. These "first opened" channels
effectively "stole" much of the flow from the capillary
beds.<o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'">*
The PFC proved virtually impossible to remove, proved obstructive to subsequent
perfusion and precluded any access to the circulation at storage
temperatures.<o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'">*
PFC perfusion was logistically very difficult and there were many problems with
cryoprotective perfusate continuing to stream out of the animals'
circulatory systems requiring extensive filtration and multiple trap placement
in the PFC perfusion circuit (on both the venous and the arterial
side).<o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"><o:p> </o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 0pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'">Perfusion
circuit of a dog undergoing subzero PFC perfusion cooling <SPAN style="mso-spacerun: yes"> </SPAN>to ~<SPAN style="mso-spacerun: yes"> </SPAN>-70 deg C in 1995:<o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'"><o:p> </o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; mso-no-proof: yes"><IMG SRC="cid:X.MA1.1378530754@aol.com" border=0 width=288 height=443 DATASIZE="26199" v:shapes="Picture_x0020_3" ID="MA1.1378530754" ></SPAN><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'"><o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'">Prototype PFC
perfusion cooling and re-warming machine developed at 21CM circa
1996:<o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; mso-no-proof: yes"><IMG SRC="cid:X.MA2.1378530754@aol.com" border=0 width=298 height=456 DATASIZE="17631" v:shapes="Picture_x0020_2" ID="MA2.1378530754" ></SPAN><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'"><o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'">And lastly, that
pumping PFC through Silastic (silicone rubber) tubing caused the build-up of
dangerous electrostatic charges which, on two occasions, resulted in fires in
the operating room and the destruction of the costly centrifugal pumps <SPAN style="mso-spacerun: yes"> </SPAN>being used to pump the PFC. One fire had
to be extinguished with a CO2 fire extinguished and it caused nearly $2,500
worth of damage - not to mention ruining the experiment! These problems were
never definitively solved...<o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'">Still, I could not
get through to <SPAN style="COLOR: black">Van Sickle and I did not receive any
response from Tanya Jones regarding this matter.<o:p></o:p></SPAN></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black">Shortly
after Peter Thiel funded Van Sickle and Jones' Argo Biomedical in February,
2012, I received an email (anonymous) informing me that the technology Argos was
developing was being kept "top secret" in large measure to avoid a response from
me, like this one. I was informed that the basis for Argos' research venture was
the communications from me to </SPAN><SPAN class=st1><SPAN lang=EN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: #444444; mso-ansi-language: EN">Van
Sickle and Jones in the mid-2000s. This prompted me to place a call to
Dr. Greg Fahy and to ask him if he knew what Argos' research platform was?
His response was that he had agreed to keep the matter confidential and that he
intended to honor that agreement.<o:p></o:p></SPAN></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt"><SPAN class=st1><SPAN lang=EN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: #444444; mso-ansi-language: EN">The
idea of using helium (or other) gas perfusion to increase both the rate <EM>and
the homogeneity</EM> of cooling in organs is not a new one and it did not
originate with me. While I can fairly take credit for being the first to propose
this for use in cryonics, and to be the first to actually apply the
technique to isolated heads, I have always been careful to credit the
researchers who originated and first validated the utility of this idea. To
those who would say that this idea is both obvious and compelling for
application in cryonics, I would point to the correspondence reproduced below,
wherein as recently as 2008, I am arguing for adoption of this approach - to no
avail.<o:p></o:p></SPAN></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt"><SPAN class=st1><SPAN lang=EN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: #444444; mso-ansi-language: EN">Reaching
the minimum concentration of cryoprotectants needed to achieve vitrification
(CNV) uniformly in human cryonics patients is very difficult due to peri- and
post-cardiac arrest ischemia. The result is that substantial freezing occurs in
areas of the brain that are not quite at CNV. The obvious way to solve this
problem is to eliminate ischemia. But, alas, this is not now possible. However
CNV is a function of not just the CPA concentration, but also of the <I style="mso-bidi-font-style: normal">cooling rate</I>. Currently, Alcor is
limited to a cooling rate of ~ -3 deg C </SPAN></SPAN><SPAN class=st1><I style="mso-bidi-font-style: normal"><SPAN lang=EN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: red; mso-ansi-language: EN">per
hour</SPAN></I></SPAN><SPAN class=st1><SPAN lang=EN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-themecolor: text1; mso-ansi-language: EN">
for brain and this means that the CNV must be very high - well over 60% (v/v).
If that cooling rate could be increased to even 0.5 deg C/min, the CNV would be
lower and more of the brain in ischemic patients would thus likely undergone
vitrification, as opposed to freezing.<o:p></o:p></SPAN></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt"><SPAN class=st1><SPAN lang=EN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-themecolor: text1; mso-ansi-language: EN">This
is what I argued be done for Alcor patients many years ago. I subsequently
repeated this argument to people at CI in 2007-8 - again to no
avail.<o:p></o:p></SPAN></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt"><SPAN class=st1><SPAN lang=EN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-themecolor: text1; mso-ansi-language: EN">In
closing I would like to quote Isaac Newton in a letter to Robert Hooke on 15
February 1676:<o:p></o:p></SPAN></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt"><SPAN class=st1><I style="mso-bidi-font-style: normal"><SPAN lang=EN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-themecolor: text1; mso-ansi-language: EN">"If
I have seen further it is by standing on ye sholders of
Giants."<o:p></o:p></SPAN></I></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt"><SPAN class=st1><SPAN lang=EN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-themecolor: text1; mso-ansi-language: EN">For
the context of this quote see:
http://www.newtonproject.sussex.ac.uk/view/texts/normalized/OTHE00101<o:p></o:p></SPAN></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt"><SPAN class=st1><SPAN lang=EN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-themecolor: text1; mso-ansi-language: EN">Generally,
(but sadly, not always) it is in the character of innovators and academics to
generously acknowledge the intellectual lineage of their work. Indeed, one of
the reasons that Newton is, to this day, so much more beloved and respected than
Robert Hooke, is because he had both the wisdom and the personal integrity to
acknowledge that his efforts, Herculean though they were, were possible only
because he himself had stood upon the shoulders of
giants.<o:p></o:p></SPAN></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt"><SPAN class=st1><SPAN lang=EN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-themecolor: text1; mso-ansi-language: EN">Mike
Darwin<o:p></o:p></SPAN></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt"><SPAN class=st1><SPAN lang=EN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-themecolor: text1; mso-ansi-language: EN"><o:p> </o:p></SPAN></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt"><SPAN class=st1><SPAN lang=EN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-themecolor: text1; mso-ansi-language: EN"><o:p> </o:p></SPAN></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt"><SPAN class=st1><SPAN lang=EN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: #444444; mso-ansi-language: EN"><o:p> </o:p></SPAN></SPAN></P>
<DIV class=MsoNormal style="TEXT-ALIGN: center; MARGIN: 0in 0in 0pt" align=center><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'">
<HR align=center SIZE=2 width="100%">
</SPAN></DIV>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt; mso-margin-top-alt: auto; mso-margin-bottom-alt: auto"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'">From:
M2darwin<BR>To: REDACTED<BR>CC: wowk@21CM.com, gfahy@21CM.com,
aschwin.de.wolf@gmail.com, chana.de.wolf@gmail.com,
m2darwin@googlemail.com<BR>BCC: danila.medvedev@gmail.com<BR>Sent: 10/4/2008
4:29:55 A.M. Pacific Daylight Time<BR>Subj: Cold Gas
Perfusion<o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt; mso-margin-top-alt: auto; mso-margin-bottom-alt: auto"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt; mso-margin-top-alt: auto; mso-margin-bottom-alt: auto"><SPAN style="FONT-SIZE: 12pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'">Hello
All,</SPAN><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"><o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt; mso-margin-top-alt: auto; mso-margin-bottom-alt: auto"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt; mso-margin-top-alt: auto; mso-margin-bottom-alt: auto"><SPAN style="FONT-SIZE: 12pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'">After
much effort I retrieved my 1970s photocopy of Pegg's paper wherein he fails
to reduplicate Guttman's work. The useful thing about this paper and the
Schimmel paper on ultra cold gas perfusion of dog and rat kidneys is that they
show what kind of cooling rates are possible and what the range of gas flows
through dog kidneys was in Pegg's hands (i.e., a credible, solid researcher
whose work is reproducible).<o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt; mso-margin-top-alt: auto; mso-margin-bottom-alt: auto"><SPAN style="FONT-SIZE: 12pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'">Schimmel
<A href="http://www.ncbi.nlm.nih.gov/pubmed/5872343"><SPAN style="COLOR: blue">http://www.ncbi.nlm.nih.gov/pubmed/5872343</SPAN></A>:<o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt; mso-margin-top-alt: auto; mso-margin-bottom-alt: auto"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'; mso-no-proof: yes"><IMG SRC="cid:X.MA3.1378530754@aol.com" border=0 width=542 height=397 DATASIZE="21838" v:shapes="Picture_x0020_14" ID="MA3.1378530754" ></SPAN><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"><o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt; mso-margin-top-alt: auto; mso-margin-bottom-alt: auto"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'">Pegg<o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt; mso-margin-top-alt: auto; mso-margin-bottom-alt: auto"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'; mso-no-proof: yes"><IMG SRC="cid:X.MA4.1378530754@aol.com" border=0 width=625 height=682 DATASIZE="51973" v:shapes="Picture_x0020_15" ID="MA4.1378530754" ></SPAN><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"><o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt; mso-margin-top-alt: auto; mso-margin-bottom-alt: auto"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt; mso-margin-top-alt: auto; mso-margin-bottom-alt: auto"><SPAN style="FONT-SIZE: 12pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'">I've
attached both papers plus a bit of history from Greg circa 1978 when he was very
hot to trot for cold gas cooling <IMG SRC="cid:X.MA5.1378530754@aol.com" border=0 width=1 height=1 DATASIZE="73" v:shapes="_x0000_i1026" ID="MA5.1378530754" >. I've also extracted and
relabeled the cooling curve graph from the Pegg paper. The principal problem
with Pegg's paper is that the kidneys were in a cold air bath; in fact her had
to wrap them in a rubber glove to SLOW the rate of cooling of the cortex.
Schimmel et al., cooled their kidneys in an insulated container with the only
source of cooling being the intra-arterial helium they used. </SPAN><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"><o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt; mso-margin-top-alt: auto; mso-margin-bottom-alt: auto"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt; mso-margin-top-alt: auto; mso-margin-bottom-alt: auto"><SPAN style="FONT-SIZE: 12pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'">Helium
flow rates through dog kidneys were 750 ml/min = or - 140 ml/min and cooling
rate to -80 deg C using gas chilled to -90 deg C was 2.8 deg C/min! This is in
close agreement with Schimmel et al's data also copied below. I've marked
up the Pegg data showing very conservative estimates for cooling and they come
out where Pegg said they do: it is important to realize that the OVERALL rate of
cooling is probably much slower than would be the case with a kidney or a
patient loaded with a vitrifiable mixture because:</SPAN><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"><o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt; mso-margin-top-alt: auto; mso-margin-bottom-alt: auto"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt; mso-margin-top-alt: auto; mso-margin-bottom-alt: auto"><SPAN style="FONT-SIZE: 12pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'">1)
There is a huge 'loss' of efficiency in cooling rate in kidneys that freeze
because the latent heat of fusion must be dissipated. This is not a problem in
systems to be vitrified.</SPAN><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"><o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt; mso-margin-top-alt: auto; mso-margin-bottom-alt: auto"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt; mso-margin-top-alt: auto; mso-margin-bottom-alt: auto"><SPAN style="FONT-SIZE: 12pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'">2)
The delta decay is enormous in Pegg's study because he held his cold gas
temperature to no lower than around -80 deg C and kept his delta of
gas to organ at around -40 to -50 deg C for the first part of the procedure
(less later on). If the delta T were TWICE this from the start the cooling rate
would be a lot faster. In fact his peak rates were in the range of 4.5 deg C min
according to the text in his paper.</SPAN><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"><o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt; mso-margin-top-alt: auto; mso-margin-bottom-alt: auto"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt; mso-margin-top-alt: auto; mso-margin-bottom-alt: auto"><SPAN style="FONT-SIZE: 12pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'">3)
He used helium which as he notes is particularly bad at heat transfer. To what
extent this made up by better flow (he reports helium literally pouring out of
the uninjured surface of the kidney!!!!! is unknown. However, if nitrogen is as
much better you guys say, then cooling rates of ~3 deg C/min should be easily
achievable for the kidney and presumably for the brain in patients without bad
vascular obstruction.</SPAN><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"><o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt; mso-margin-top-alt: auto; mso-margin-bottom-alt: auto"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt; mso-margin-top-alt: auto; mso-margin-bottom-alt: auto"><SPAN style="FONT-SIZE: 12pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'">If
you eliminate the problem Pegg had with the superficial cortex cooling
faster than the deep cortex and medulla by insulating the patient/organ
from the cold bath gas then you would see a virtual abolition of any significant
difference between surface and core cooling - presumably eliminating
viscoeslastic injury.</SPAN><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"><o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt; mso-margin-top-alt: auto; mso-margin-bottom-alt: auto"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt; mso-margin-top-alt: auto; mso-margin-bottom-alt: auto"><SPAN style="FONT-SIZE: 12pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'">4)
Finally, unlike the case with PFCs the vasculature will be open so that
nanomachines or worst case, fixatives, can be introduced later at subzero
temperatures. It will also be far cheaper than using PFCs if N2 works
OK.</SPAN><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"><o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt; mso-margin-top-alt: auto; mso-margin-bottom-alt: auto"><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"> <o:p></o:p></SPAN></P>
<P class=MsoNormal style="MARGIN: 0in 0in 10pt; mso-margin-top-alt: auto; mso-margin-bottom-alt: auto"><SPAN style="FONT-SIZE: 12pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'">Mike
Darwin</SPAN><SPAN style="FONT-SIZE: 10pt; FONT-FAMILY: 'Arial','sans-serif'; COLOR: black; mso-fareast-font-family: 'Times New Roman'"><o:p></o:p></SPAN></P>
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