X-Message-Number: 2874
Date: 09 Jul 94 03:02:55 EDT
From: Mike Darwin <>
Subject: CRYONICS Case Report (3/3)

  -- Continued --

Total Body Washout (TBW)

	TBW was begun at 17:28 using 8 liters of Viaspan at a flow 
rate of 630 cc/min, MAP of 50 mmHg, oxygen flow of 4 LPM, 
arterial temperature of  8.7 C, esophageal temperature of 6.1 C 
and a rectal temperature of  16.8 C.  The composition of the 
Viaspan with additives to it are given in Table I.  TBW was 
concluded at 17:49 at a flow rate of 630 ml/min, MAP of 50 mmHg  
and an esophageal and rectal temperature of  4.8 C and 15.2 C 
respectively.  Oxygen flow-rate to the oxygenator was reduced to 
2 LPM at the conclusion of  TBW.

Table V: Composition of Viaspan

Component			         g/l

Lactobionic Acid			        50.0
Potassium Phosphate, monobasic	        03.40
Magnesium Sulfate, heptahydrate	        01.23
Raffinose, pentahydrate		        17.83
Adenosine			        01.34
Allopurinol			        0.136
Glutathione			        0.922
Dexamethasone*			        0.16
Glucose*			        0.90
Insulin (U-100, Regular, Human)*	        40 IU
Heparin*			      2000 U/L
Water for Injection	                        q.s.
Potassium Hydroxide		         q.s.
Sodium Hydroxide		   adjust pH to 7.4
Measured Osmolality: 305 mOsm/kg
Measured pH: 7.5

*Denotes component added immediately before use.

	Recirculation  of Viaspan was continued until 20:04 at which 
time extracorporeal support was discontinued.  At the conclusion 
of bypass the patient's esophageal temperature was 1.8 C and the 
rectal temperature was 6.2 C.   The lines to the femoral cannula 
were  clamped and cut approximately 10 cm from where they were 
connected to each cannula.  A 1/2" -3/8" connector was used to 
connect the stubs of the arterial and venous lines together 
(carefully avoiding the introduction of air) effectively cross-
connecting the arterial and venous lines.  The occluding clamps 
were then removed and the wound (with cannula in-place) was 
packed with sterile 4" x 4" gauze and covered with a 3M Steri-
Drape adhesive dressing.

	The patient was then cleaned up in the PIB, transferred from 
the PIB into a heavy-duty vinyl body bag, and the head rapidly 
repacked in ice using 1-gallon Zip-Loc bags filled with shaved 
ice.  The body bag was closed and the patient was carried in the 
body bag (using the body bag as a stretcher) down the steep 
flight of steps from his condominium to the PIB on the Mobile 
Advanced Life Support System in the parking lot.  (The MALSS PIB 
had been previously prepared by laying a layer of Zip-Loc bags 
filled with shaved ice  in the bottom of it) Once the patient was 
in place in the PIB of the MALSS the body bag was opened and the 
patient was thoroughly packed in ice from head to foot using Zip-
Loc bags filled with shaved ice.  The PIB cover was placed and 
the patient was wheeled into the BPI ambulance for ground 
transport from Sunnyvale, CA where cardiac arrest had occurred to 
BPI's cryoprotective perfusion facilities in Rancho Cucamonga, 

	The patient arrived at the facility in Rancho Cucamonga at 
06:30 with an esophageal temperature of  1.0 C and a rectal 
temperature of 0.6 C.

Gross Assessment
     The patient was positioned on the operating table at 
approximately 09:50.  After the patient was positioned on the 
operating table, he was briefly examined.  Initial assessment of 
the patient revealed a markedly dehydrated (globes deeply 
retracted) cachectic male with complete rigor (except in the left 
leg).    The eyes were opened and the eye exam disclosed 
bilaterally dilated pupils with corneal misting.    The skin was 
pale, bloodless in appearance, uniform in color, of markedly 
decreased  turgor, and free of lividity in dependent areas.  The 
skin/sternum under the Thumper piston was deeply indented and 
bruised in appearance as is typical in patients subjected to 
prolonged  Thumper support.  

Perfusate Preparation

     The composition of the perfusate employed to carry out 
cryoprotective perfusion is given in Table VII.  Dry chemical 
perfusate components were prepared from reagent or medical grade 
chemicals  using an Ohaus Model TS400D electronic balance 
(previously weighed out and stored under dessicated conditions).  
Dry components were mixed with American Chemical Society (ACS) 
reagent grade glycerol and sterile water for injection USP, 
and/or sterile water for irrigation USP.  Perfusates were 
sterilized by filtration into the recirculating and 
cryoprotective concentrate reservoirs through a Pall PP3802 0.20u 
pre-bypass filter.   Perfusate was prepared in two batches with 
the following volumes and glycerol concentrations:

Table VI: Perfusate Volumes and Glycerol Concentrations

Description                Volume       Glycerol%(v/v)
 Recirculating              40 liters               5%

Concenrtrate                60 liters            65%


Mannitol-HEPES-2 (MHP-2) Perfusate Composition

Component                  Molar Concentration (mM)             

Mannitol                       170 (MW 182.20)                         

Adenine HCl                  0.94 (MW 180.6)                         

D-Ribose                       0.94 (MW 150.2)                         

Sodium Bicarbonate      10.0 (MW 84.0)                            

Potassium Chloride        28.3 (MW 74.56)                         

Calcium Chloride           1.0  (MW 111)                  0.28 
ml  10% soln.

Magnesium Chloride      1.0  (MW95.2)                  1.0 ml  
20% soln.

HEPES (Na+ salt)         15.0 (MW 260.3)                         

Glutathione                     3.0  (307.3)                                  

Glucose                         5.0  (180.2)                                   

Hydroxyethyl Starch          ---                                            

Heparin                             ---                                       
1000 units/l

	Analysis of the perfusate with the Nova Stat-5 Blood 
Gas/Electrolyte Analysis System disclosed the following:

pH = 8.202
Na+ = 72 mM/l
K+ = 24.1 mM/l
Cl- = 83 mM/l
Ca++ = 0.40 mM/l
Glucose = 77 mg/dl
mOsm/kg = 400

Operative Procedures

Pre-operative Prep

     The patient was prepared for a median sternotomy and cranial 
burr-hole by shaving the head and thorax and scrubbing/swabbing 
them with povidone iodine solution (Betadine).  The sternal 
operative site was defined by draping with sterile muslin towels.  
A fenestrated cardiac drape was placed over the patient, "tented" 
on two IV poles at the head, and allowed to extend down over the 
feet and over the sides of the table by a minimum of 24".  The 
top of the scalp was draped with a fenestrated adhesive drape 
over the left frontal lobe.

Median Sternotomy/Vascular Access

     Median sternotomy was begun at 12:04 on 02-06-1994 with an 
incision over the midline of the sternum with an electrosurgical 
knife.  Fascia and connective tissue were cleared down to the 
sternum with the electrosurgical knife.  A median sternotomy was 
then performed with a Stryker oscillating sternal saw.  The edges 
of the sternotomy were padded with laparotomy sponges, a self-
retaining retractor placed, and the sternotomy retracted open.  
The left pleural space was opened and blunt and sharp dissection 
were used to expose the pericardium.  A ventral midline 
pericardiotomy was made using Metzenbaum scissors.    A Sarns 
cardiotomy sucker was used to suction away the pericardial fluid 
to discard.

	Braided polyester purse string sutures (2-O Ti-cron) were 
placed in the right atrial appendage and two were placed in the 
aortic arch.  A 5 Fr. Argyle polyethylene cannula was placed in 
one of the purse-string sutures to monitor arterial pressure.  A 
24 Fr. x 40 cm USCI type 1967 Venous cannula was placed in the 
right atrial appendage and passed into the right atrium.  The 
caudal vena cava was ligated with #1 silk.  The descending 
thoracic aorta at the level of the 4th rib was isolated and 
doubly ligated with umbilical tape. A 16 Fr. Sarns aortic 
perfusion cannula was primed with normal saline and a clamp 
placed on the distal end.  The cannula was then introduced into 
the aorta and anchored  in place with the purse string suture.  
All cannulae were secured with # 1 silk suture to each line.

	Industrial/automotive stainless steel hose-clamps were used 
as tourniquets and were placed on each of the patient's arms at 
the level of the axilla to achieve non-surgical vascular 
isolation of the upper appendages.  

              The sterile perfusion tubing was then brought up to 
the surgical field and secured in a Travenol tubing holder towel-
clamped to the drapes.  The arterio-venous loop of the perfusion 
circuit was clamped and divided by cutting out the 1/2"- 3/8" 
adapter with Mayo scissors.  A 1/2" connector with a Cobe 4-way 
stopcock was used to connect the 1/2" ID venous return line to 
the venous cannula.  Air was cleared from the system with a 100 
cc glass syringe.  A Cobe 8 ft. pressure monitoring line was 
fitted to the arterial pressure catheter, flushed with normal 
saline, and handed off the sterile field to be connected to a 
Trantec Model 800 pressure transducer and a Tektronix Model 414 

     Surgery to connect the patient to the perfusion circuit was 
completed at 13:29.  
Cranial Burr-Hole

	Surgery to open the cranial burr-hole was begun at 12:12.  
The vertex of the scalp approximately 3 cm to the right of 
midline over the right frontal lobe was incised with a #10 
scalpel blade and an incision approximately 4 cm long was made 
down to the periosteum.  A periosteal elevator was used to expose 
the parietal bone approximately 3 cm to the right of the midline.  
A 10 mm hole was made with a DePuy pneumatic perforator.  The 
burr-hole was then increased in diameter using a Hudson brace and 
10 mm neuro burr.  The dura mater was left closed and the 
cortical surface was palpated through the dura and determined to 
be about 3 mm below the inner aspect of the cranial vault  The 
dura appeared blood free.  

Figure 4: Burr-hole location.

	Opening of the dura was deferred until 14:43  in order to 
attempt to determine the source of  the copious drainage normally 
observed coming from the burr hole during cryoprotective 
perfusion of patients. Drainage from the scalp and bone wounds 
was determined to be 10-15 cc/min.  Approximately  30 minutes 
after the start of perfusion the dura was again depressed and 
found to be moderately tense.  Either cerebral edema or fluid 
accumulation in the subdural space had begun to occur.  Seventy-
three minutes into cryoprotective perfusion the dura was breached 
by perforating it with a 16 g needle and a copious, moderately 
high pressure flow of clear fluid was observed to issue from the 
needle.  At this point the needle was withdrawn and the puncture 
in the dura was widened with a #11 scalpel blade to approximately 
1 mm in diameter.  A copious and pressured flow of fluid was 
observed streaming from the puncture.  A 16 g blunt needle was 
then passed through the opening in the dura and a sample of the 
fluid collected in a 3 cc syringe for analysis of glycerol 
concentration and gases and electrolytes.  The results of that 
analysis demonstrated a close correlation in composition between 
venous perfusate and subdural effluent as contrasted with the 
composition of arterial perfusate.

Table VIII: Composition of Subdural Fluid Compared with Venous 
Perfusate (14:45)

Test	    Subdural Fluid/Venous Perfusate/Arterial Perfusate

pH	       7.509                      7.518	         7.576
pO2  	        58		       66                              109.1
pCO2	        18.1	      17.4                             3.9
Refractive Index     28.2             28.4                            30.8

	Following evaluation of the subdural fluid the dura was 
elevated with a dura hook and trimmed to the margins of the burr 
hole allowing for inspection of the cortical surface and free 
drainage of fluid from the subdural space.

Perfusion Circuit 

	The extracorporeal circuit for cryoprotective perfusion 
consisted of two parts: a recirculating system to which the 
patient was connected, and a cryoprotectant addition system which 
was connected to the recirculating system.  The recirculating 
system was a 60 liter reservoir sitting atop a magnetic stirring 
table, an arterial (recirculating) roller pump, a Sarns 16310 
hollow fiber oxygenator/heat exchanger and a Pall EC1440 40 
micron blood filter.  The recirculating (mixing) reservoir was 
continuously stirred with a 2" Teflon-coated magnetic stirring 
bar driven by a Thermolyne type 7200 magnetic stirrer.  The 
cryoprotectant addition system consisted of a 60-liter 
polyethylene reservoir containing 65% (v/v) glycerol (see Table 
II) and a Drake-Willock model #7401 hemodialysis pump acting as a 
cryoprotective addition pump which added 65% (v/v) glycerol 
perfusate from the concentrate reservoir to the recirculating 
reservoir.  A Travenol 5M1153 roller pump was used to remove 
perfusate from the recirculating system via the venous return 
line and discard it to the drain. (This system is illustrated in 
Figures 5 below.)

Figure 5: Cryoprotective perfusion circuit incorporating VBL and 
Sarns 9438 reservoir.

	A floating lid was used atop the recirculating perfusate 
column to prevent air from being entrained in the perfusate by 
the stirrer bar.  A Sarns filtered (40 micron) hard-shell venous 
reservoir was interposed between the recirculating reservoir and 
the arterial pump to serve as an air trap/defoamer in the event 
foam was generated in the recirculating reservoir and also as an 
arterial prefilter for particulates (agglutinated RBCs, etc.) .

	Arterial and venous samples for evaluation of chemistries 
and glycerol concentration were drawn at 15-minute intervals 
during cryoprotective perfusion.  Arterial samples were drawn 
from a 3-way, 4-stopcock manifold interposed between the arterial 
filter and the filter vent line.  Venous samples were also drawn 
from the stopcock assembly which was connected by an 8' Cobe 
monitoring line to the venous return line approximately mid-way 
along its course to the recirculating reservoir. (The dead-space 
of the Cobe monitoring line was determined and a volume in excess 
of the dead-space volume was drawn up and held in a syringe 
attached to the middle stopcock of the Cobe manifold before each 
sample was taken.)

 	The perfusion circuit was prepared in advance of need and 
was sterilized with ethylene oxide using an appropriate protocol 
of post-sterilization out-gassing and aeration.  A blend of 
oxygen and helium gases was delivered to the oxygenator at a flow 
rate of  2-liters per minute (this sweep gas flow rate was 
maintained  throughout cryoprotective perfusion).  FiO2  to the 
oxygenator was maintained between 18-25% throughout perfusion

Cryoprotective Perfusion

 	Open-circuit perfusion of  5% (v/v) glycerol perfusate was 
begun at 13:30 at a flow rate of 1100 cc/min., an esophageal 
temperature of  2.7C,  an arterial temperature (perfusate) of 
12.0 C and a mean arterial pressure (MAP) of  61 mmHg. 
Approximately 15 liters was used to flush the patient's 
circulatory system of Viaspan and residual blood and accumulated 
metabolic byproducts from the period of cold ischemia during 
transport before the start of cryoprotective perfusion.  A 
significant amount of cold agglutination was noted during this 
initial flush (typical "grains of sand" clumps of  red cells 
which disappeared on collection and warming), but there were no 
signs of clotted material.  Additionally, it should be noted that 
whole blood samples drawn on this patient prior to and during 
transport (anticoagulated with EDTA) also showed marked cold 
agglutination upon refrigeration to 2-4 C (Agglutination Score = 
9, after Marsh, RL, Transfusion 12;5: 352-353, 1972).   At 13:30 
arterial pH and gases were as follows: pH 7.772, pO2 202.1, pCO2 
1.4 and venous pH and gases were pH 7.328, pO2 21.8, pCO2 4.5 at 
an FiO2 of 23%. The glycerol ramp was begun at the start of 
closed circuit perfusion at 13:45 at an addition/withdrawal rate 
of 200 cc/min.  Closed circuit cryoprotective perfusion was begun 
at 13:45 with 20 liters of 5% (v/v) glycerol perfusate in the 
recirculating reservoir and 40 liters of 65% (v/v) glycerol 
perfusate in the concentrate reservoir.

 	At 14:42 glycerolization of the face and scalp was noted to 
be patchy with many non-perfused areas noted (possibly due to 
cold agglutination in the skin).  The skin on the neck and upper 
thorax was observed to be glycerolizing somewhat more evenly and 
appeared to have fewer   non-perfused areas. The left ear and 
left face appeared to be glycerolizing better  than the right ear 
and face.  It was noted that the right side of the patient's head 
was the dependent side (the patient's head was turned to the 
right) during the last few hours of life.  In particular a large 
(20-30 cm) area of scalp near the occiput of the head on the 
right side was noted to be cherry red and nonperfusing -- 
apparently an infarcted area as a result of a compression of the 
skin (and resultant ischemia) during the hours of  shock and 
immobility which characterized the patient's agonal course.

Cryoprotective Ramp

	After the burr hole was opened at 14:43 withdrawal of 
perfusate from the recirculating system was temporarily 
discontinued as leakage from the burrhole was in excess of the 
desired withdrawal rate (leakage was 150 cc/min).  After 
evaluating the recirculating reservoir level for  30 minutes the 
withdrawal pump was again started at a rate of 110 cc/min.   The 
rate of increase in arterial glycerol concentration during most 
of the first third of perfusion was 46 mM/min..  This 
differential had declined to 25 mM by the second third of 
perfusion and was approximately 29 mM for the last third of 
cryoprotective perfusion..  This resulted in an average 
arterial/venous difference in glycerol concentration of 
approximately  36 mM over the course of cryoprotective perfusion.  
Glycerol concentration increase in both the arterial and venous 
perfusate are given in graphic form at the end of this report and 
in tabular form below:

Time     (Arterial M)      (Venous Glycerol M)
1352            1.13                        0.95
1400            1.43                        1.18                          
1418            1.71                        1.62
1430            3.18                        2.73
1445            3.84                        3.40
1500            4.41                        3.90
1515            4.94                        4.38
1530            5.33                        4.90
1545            5.84                        5.28
1600            6.08                       5.69
1621            6.38                       6.09
1635            7.14                       6.76
	The initial response to the start of the cryoprotective ramp 
was poor in the skin but good in the brain, with cerebral 
cortical volume rapidly decreasing to 2-3 mm below the inner 
aspect of the cranial vault.  The pial surface was observed to be 
blood free over the visible area of the right hemisphere (an area 
of  about 20-30 mm in all directions from the burr hole; the 
limits of illumination of the cortical surface with a pen-type 
flashlight). The cortical surface continued to shrink away from 
the burr hole  to a low of approximately 7 mm below the inner 
aspect of the cranial vault.  At approximately 15:23 brain volume 
was noticed to have increased slightly with the cortical surface 
at 3-4 mm below the cranial vault.  By 16:21 the cortical surface 
was at the level of the burr hole opening or perhaps .5 mm into 
the burr hole.  At 16:41 The cortical surface was approximately  
+1 mm into the burr hole at the conclusion of cryoprotective 
perfusion.  Immediately after the termination of perfusion (i.e., 
after the arterial pump was turned off) the cortical surface 
receded to 1.0 mm below the inner aspect of the cranial vault.

	At 16:22 the rate of concentrate addition to the 
recirculating system was increased from 200 cc/min to 400 cc/min 
and the withdrawal pump rate was increased from 115 cc/min to 220 
cc/min.  Perfusate levels in the concentrate and recirculating 
reservoir were 7.5L and 17L, respectively.  Arterial  flow rate 
at that time was 675 cc/min, MAP was 81 mmHg, arterial 
temperature was 14.0 C and esophageal temperature was 9.7 C.

	Blood gases, pH and electrolytes during perfusion are 
appended to the end of this case history.

	Cryoprotective perfusion was concluded at 16:35 at an 
arterial flow rate of 500 cc/min., MAP of 81 mmHg, arterial 
temperature of   11.6 C, esophageal temperature of 9.2 C,  
recirculating system addition/withdrawal rate  was 400 /220 
cc/min.   Final  venous gases/electrolytes were not measurable 
due to the high viscosity of the perfusate (the Nova Stat-5 
Profile is unable to process such viscous samples).  The last 
venous gases/electrolytes for which data wereobtainable was drawn 
at 16:00 and were as follows: pH 7.543, pO2 28.2 mmHg, pCO2 4.4 
mmHg, Na+ 104 mM, K+ 71.1 mM, Cl- 77 mM, Ca++ 0.34 mM, glucose 81 
mg/dl.  Laboratoy analysis of arterial and venous samples drawn 
during cryoprotective perfusion are presented in graphic form at 
the end of this report.

	Reservoir levels  at the conclusion of cryoprotective 
perfusion were: concentrate 1-2 L, and recirculating 7 L.  The 
final venous glycerol concentration was 6.76 M.

Burr-Hole Closure

  At 16:43, the silastic-coated tip of a 15' long, 30 gauge 
Capton-clad copper-constantan (type T) thermocouple probe 
(Instrument Laboratory #53- 30-513) was threaded into the burr-
hole and placed on the cortical surface.  The burr-hole was then 
filled with bone wax and the scalp closed with surgical staples.  
Evaluation of cerebral cortex temperature was delayed until the 
burr-hole had been closed,  at which time it was discovered that 
the thermocouple was not functional.  The burr hole was reopened 
(staples and bone wax removed) and a  new (functional) 
temperature probe was placed.  The burr hole was closed as before 
and the  probe wire was anchored to the scalp with surgical 
staples.  Brain surface temperature was measured at 9.5 C at 
17:00, esophageal temperature was 7.0 C.

	Between 17:00 and 17:12,  20 gauge,  teflon clad, copper-
constantan TC probes were placed nasopharyngeally and on the skin 
of the right temple.  Both probes were anchored to the patient by 
stapling the probe wires to the skin with surgical staples. 

Cephalic Isolation

	Surgery for cephalic isolation was begun at approximately 
17:12 by making a circumferential incision at the base of the 
neck at just above the level of the clavicle using a Black and 
Decker Slim-Grip (Catalog # EK200, Type 1)  Electric Carving 
Knife.  The Electric Knife was used to incise the cervical skin, 
musculature, and other structures down to approximately the 5th 
or 6th cervical vertebrae.  The vertebral column was then cut 
with a Satterlee amputation saw, freeing the head from the body.

	The cervical musculature (in contrast to the facial skin) 
was observed to be uniformly perfused and glycerolized as 
indicated by its stiff and waxy texture and dark (maroon) color.  
The spinal cord was observed to be blood free, slightly shrunken 
within the vertebral foramen, and otherwise giving an appearance 
consistent with good glycerolization.  The cervical vessels were 
observed to be blood free and the cervical tissues at cross-
section appeared to be free of infarcts and evenly glycerolized.

	The  cephalic cervical stump was covered with sterile gauze 4" x 4" s 
which were in turn covered and held in place with a sterile 
Stockinet.  Cephalic isolation was completed at 17:20

Cooling to Dry Ice Temperature

	Temperature descent to -79 C was monitored with probes in/on 
the pharynx,  brain surface and head surface (placed temporally).  
An additional probe was used to monitor bath temperature.  Bath, 
external, and nasopharyngeal probes were Instrument Laboratories 
53-20-507, "load type", 20 gauge, Teflon-coated copper-constantan 

	The patient (cephalon) was placed inside two 1.4 mil 
polyethylene plastic bags and lowered into a 20 liter bath of 5 
centistoke polydimethylsiloxane oil (Silcool) which had been pre-
cooled to -22 C with dry ice.  The first temperature readings 
taken at 17:24 were: brain surface 7.0 C, temporal 8.7 C, 
nasopharyngeal 7.5 C and Silcool bath - 21.4 C.  Controlled rate 
cooling was achieved by the manual addition of dry ice to the 
Silcool bath.

	Cooling to -77 C was at a rate of approximately 5 C per hour 
to -40 C, and approximately 4.5 C  per hour from -40 C to -77 C.  
Cooling to - 77 C was complete by 10:45 on 02-07-1994.  Patient 
cooling data to -79 C are presented tabularly and graphically as 
an addendum to this document.

	Cooling was achieved by addition of dry ice to the bath per 
the following protocol:

1) Hold bath temperature at -40 C until nasopharyngeal 
temperature reaches - 35 C.
When nasopharyngeal temperature reaches -35 C decrease bath 
temperature to -50 C.

3) Hold bath temperature at -15 C below nasopharyngeal 
temperature to maintain a bath-to-core temperature differential 
of 15 C.

4) When an nasopharyngeal temperature of -65 C is reached add 
sufficient dry ice to take the bath temperature down to -79 C and 
hold it there. 

	Once cooling to -79 C was complete the patient was lifted 
out of the Silcool bath, the outer, Silcool wetted plastic bag 
was stripped off and the patient was placed within a polyester 
cloth bag which was in turn placed inside a standard aluminum 
neurocan which had been precooled to -90  and packed with dacron 
wool.  Temperature probes were left externalized to the neurocan 
for subsequent cooling to -196 C.

Serum/Perfusate Electrolytes and  Enzymes

	Laboratory evaluations of samples taken during 
cryoprotective perfusion are presented in full in tabular and 
graphic form as an addendum to this document.  All samples were 
analyzed with a Nova Stat Profile 5 (in-house) or by SmithKline 
Beecham Clinical Laboratories of  VanNuys, California using an 
Olympus AU5061 Clinical Chemistry Analyzer.

	Final venous chemistries were as follows:

Na+				46 mM/L
K+				28.4 mM/L
Cl-				16 mM/L
Ca++				0.9 mg/dl
Glucose 				74 mg/dl
Urea Nitrogen			7 mg/dl
Creatinine			0.2 mg/dl
Phosphorus			2.2 mg/dl
Protein, Total			1.5 g/dl
Albumin			0.5 g/dl
Globulin, Total			1.0 g/dl
A/G Ratio     			0.5
Bilirubin, Total                                      0.1 mg/dl
Alkaline Phosphatase	                3 U/L
Lactate Dehydrogenase                      211 U/L
GGT				 2 U/L
AST				 72 U/L
ALT			                 18 U/L
Uric Acid			0.3 mg/dl
Triglycerides			1121 mg/dl
Cholesterol, Total		1 mg/dl

 Michael Darwin, Cryopreservation Team Leader            Date

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