Members have access to national West Nile reactive donation data by state, and by donor residential zip code, via the AABB Biovigilence Network, and by way of communication plans established by blood organizations.  West Nile activity, as observed from West Nile RNA-confirmed- positive blood donors, occurs from the beginning of May through November.  Available data indicates that the use of ID-NAT during these times of West Nile activity is implemented by either single facilities or multiple facilities representing overlapping and adjacent areas, will greatly reduce the risk of transfusion-transmitted West Nile virus.

            During 2007, no cases of transfusion-transmitted West Nile virus were reported.

            Recommendations put forward in AABB Association bulletins regarding West Nile are developed based upon available data that are reviewed by the AABB's West Nile Task Force, which includes representation from the national blood organizations and obtains input from liaison representatives from the FDA and CDC.

            Such recommendations are also reviewed by AABB's Transfusion-Transmitted Diseases Committee, which has broad representation from national and international experts.

            Lastly, all AABB Association bulletins are approved, prior to release, by the AABB board of directors.  One of the objectives of the association bulletin developed for 2007 was to validate the recommended minimum trigger, which was based on the absolute number of West Nile reactive donations in a given geographic area as well as the rate of West Nile reactive donations in that same area over a seven day rolling period.

            The validation data, which was collected by a prospective study conducted by the American Red Cross, in six regions having experienced recurrent West Nile activity, revealed that 25 of 30, or 83 percent of confirmed positive donations were not detected using the recommended trigger.

            Increased sensitivity could be achieved with the elimination of the rate criterion and the use of only the absolute number of West Nile reactive donations.

            Prior to the release of revised recommendations regarding the use of a new trigger--that's for this year--agreement was reached that the use of presumed viremic donations or PVDs--those are donations with elevated signal-to-cutoff values, or those that are reactive--should be used in place of initially reactive donations for triggering and detriggering determinations.

            This is the practice that has been used successfully by blood centers for the past several years.

            In contrast to initially reactive results, PVDs have a sensitivity and positive predictive value of greater than 95 percent.  So that as decisions to convert from minipool to ID-NAT are made, resources, including both labor and reagents, are used wisely and focused in the areas of greatest need.  The false-positive rate of the NAT assays, relative to initially reactive results of one to two per thousand tests--that's the Red Cross experience, which is lot-dependent--would result in unnecessary triggering events, but, more importantly, would make it difficult or impossible to ever transition back to minipool NAT.

            The validation data revealed that 56 of 57, or 98 percent of donors testing falsely positive, occurred during West Nile ID-NAT, and therefore, more specific criteria than the use of initial reactivity for determining conversion from minipool to ID-NAT, and back to minipool NAT are needed.

            The West Nile Task Force reviewed three options for triggering based on the validation data and elimination of the rate criterion.  The three options reviewed prior to the release of the association bulletin included one PVD, not an initially-reactive donation with a rate determination, two PVDs, again without a rate determination, and thirdly, a hybrid approach which included a blending of one and two PVDs, again without a rate determination.

            One PVD would be used in predetermined areas of West Nile activity and two PVDs would be used in areas that have never experienced previous West Nile activity.

            The intent, again, was to carefully evaluate where the need was greatest and not falsely trigger in a given area such that labor reagents, donors and their donations would be conserved.

            Following consideration of the above three options, the decision was made to recommend two PVDs within a seven day roll-in period without a rate requirement.  Facilities are encouraged to review the validation data provided in the association bulletin and make local triggering and detriggering decisions as appropriate.

            However, it is recommended that facilities consider the benefits of establishing uniform criteria.  Facilities are also encouraged to review carefully location conditions, and that triggering on one PVD is appropriate.

            In the event that facilities in overlapping or adjacent areas have PVDs or local conditions indicate ongoing West Nile activity, such as reported clinical cases or reports of positive birds or mosquito pools, detriggering is based on seven days without a PVD, ID-NAT should continue beyond seven days in areas with ongoing West Nile activity, including positive blood donors from facilities that collect in overlapping areas or if other conditions such as prior history, ongoing clinical avian or mosquito activity exist, or at the discretion of the medical director.

            In these situations, ID-NAT for 14 days should be considered.

            The new triggering and detriggering criteria are believed to represent an approach that is believed to reduce the already low risk of West Nile transfusion transmission.

            Lastly, the AABB and other professional organizations, and blood collecting organizations, will be responding with comments to the FDA draft guidance.  Thank you.

            DR. SIEGAL:  Thank you very much.  All right.  At this point we'll adjourn for lunch, but I ask the committee to come forward to talk a little bit about the new voting procedures.  We'll resume at 1:30.

            (Whereupon, a luncheon recess was taken, the committee to resume at 1:30 p.m., the same day.)

            DR. SIEGAL:  Okay.  Let's come to order, and we'll start right away with topic one, relating to the BEST study, and the committee report on red blood cell recovery standards.

            We're first going to hear from Ping He, M.D., the medical officer at FDA, on FDA's approaches to evaluation of red cell products.

            Dr. He.  Have I pronounced that correctly?

            DR. HE:  Good afternoon.  My name is Ping He.  I am from Division of Hematology at Office of Blood Research and Review, and this afternoon, I'm going to talk about the FDA's criteria for evaluation of red blood cell products.

            Here's the issue summary for what we are going to discuss this afternoon.  FDA seeks the advice of the committee on an industry proposal to change the current acceptance criteria for evaluation of red blood cell studies based on in vivo radiolabeling recovery trials.

            Here's the key issue that we are going to talk about this afternoon.  Should the RBC in vivo recovery threshold value be changed?  The threshold value of the greater than equals 75 percent for RBC recovery study, serve as a cutoff line for determining RBC viability for unit during the evaluation of RBC in vivo recovery studies.

            The threshold value of greater than equals 75 percent has been--oh.  Okay.             The threshold value for--the threshold value of greater than equals 75 percent has been used by FDA in the past 24 years for evaluation of RBC in vivo recovery studies, based on the expert opinion and historic data.

            And this diagram shows that if a unit of RBC has in vivo recovery greater than 75 percent, that it was considered as a successful unit, and that means that the minimum amount of RBC, viable RBC in this unit would be greater than 75 percent.

            However, recently, the industry did a study based on the historic data, and indicated that the overall RBC recovery studies from 1990 to 2006 would not meet the FDA's current acceptance criteria--I'm going to talk about later--unless the threshold value, it changed from 75 percent to 67 percent, meaning that the viable RBCs in each bag should be dropped to 67 percent, while in contrast to the industry analysis, based on the more recent data, FDA believes that RBC products are improving with time.

            During the period 1998 to 2007, 17 out of 19 RBC recovery studies met the current acceptance criteria.

            Therefore, FDA maintains that the threshold value of greater than equals 75 percent should not be changed.

            We all know that RBC products play an important role in transfusion medicine.  RBCs are life-saving products that deliver oxygen to tissue.

            Each year, about 14 million units of full blood are collected, and on a given day, more than 38,000 units of RBC products are needed for patients with anemia, trauma, cancer or surgical procedures.  Up to now, there is no available substitute for RBC products, and the demand for RBC products are continuously growing.

            However, the collection or processing or manufacturing of RBC products may cause storage lesion.  A unit of RBC products can be collected either through the whole blood collection procedure which goes through steps of centrifugation and separation, or can be collected through the complex procedure.

            And then RBC products can be stored at different anticoagulants or additive solutions at 1 to 6 degree for a maximum shelf life of 42 days.  Studies have been showing that any preservation or manipulation can induce RBC membrane damage, therefore producing changes in the biochemical properties of RBCs and shortening their in vivo survival.

            Here, the potential harmful effects of the RBC storage lesion from the practical view, the most important change of RBC storage is the loss of RBC viability, and therefore shortens time in circulation after transfusion into the recipient.

            It therefore will decrease the oxygen delivery to tissue and the transfusion of damaged RBCs to recipient may saturate macrophage clearance mechanisms, and therefore reduce the bacterial clearance.

            The storage lesion can also decrease the levels of ATP in RBC, which correlates with the reduced RBC viability, and decrease the tissue perfusion.  Decrease 2,3-DPG, reduce oxygen delivery to tissue.  Loss cell membrane can make RBCs rigid, and therefore increase the spontaneous lysis, decrease microvascular flow.

            Storage lesion can also cause increase in hemolysis which has a harmful effect on organ function, and increased plasma potassium can cause increased potential hazard to neonates.

            Therefore, in order to ensure a new device for presence of RBC products, safe and effective, here, the FDA recommended tests on approval/clearance of stored processed RBC products.  Basically, a unit of the whole blood will be collected from a healthy volunteer into this new bag, and then the blood will be stored for 42 days, and we know that at end of the storage some of the RBC becomes nonviable.  Now in order to determine the levels of viable cells in a stored RBC bag, a portion of RBC will be collected from the storage bag and radiolabeled, and reinfused back to the same donor, and the 24 hour RBC in vivo recovery study will determine the viability of the RBCs.

            Meanwhile, a panel of the individual RBC test will also be determined from the stored RBC product, such as ph, hemolysis, levels for ATP, 230 pg, hemoglobin, hematocrit, cell morphology counts and glucose, and so on.

            However, the RBC individual test are used as screening tests, and the individual test are not predictive of in vivo RBC performance.  Therefore, the 24 hour RBC in vivo recovery remains critical for evaluation of effectiveness of a new bag.

            So the in vivo RBC recovery at 24 hours provides a surrogate end point for RBC product to evaluation.  It is to demonstrate that the safety and effectiveness of novel RBC collection processes --

            This diagram shows the comparison of RBC in vivo recoveries with different RBC storage period.  It was published by Mollison in 1951.

            These three lines shows the post-transfusion red blood cell survival from fresh blood, of red blood cell stored for 14 days or stored for 28 days in ACD anticoagulant at 4 degrees, and here the axis shows the days of the transfusion, the Y axis shows the percent of the RBC survival.

            I would like to draw your attention to the 24 hour post-transfusion time point, showing that the 24 hour post-transfusion recovery for the fresh blood is great, about a 100 percent.  It drops to 95 percent when the cell is stored at 14 days, and it drops to 75 percent when the cell is stored for 28 days.

            This means that the longer the storage, the poorer the RBC survival and recovery.  As I had mentioned earlier, that the greater than, or equals 75 percent of RBC recovery at 24 hours, is a threshold value for individual unit recovery, that it has been used for evaluation for in vivo recoveries.

            Now in order for FDA to determine if a clinical study is successful, certain criteria has used in the past, and this is a historical review of the FDA acceptance criteria for evaluation for in vivo RBC studies.  Basically, we recommend the study should be done in more than two different centers, with a minimum of twenty healthy volunteers.

            In 1985, at the FDA workshop on red cells stored in additive solution, based on the historic data and expert opinion, the acceptance criteria for RBC in vivo recovery started out with a mean recovery of  multiple individual units greater than equals 75 percent, meaning that if you have a sample size of twenty, regardless, the recovery ranged from 30, 40 percent to 80, 90 percent.  Or regardless of individual units that has recovery less than 75 percent, three out of twenty or five out of twenty, as long as the average or mean recovery of 20 units, greater than equals 75 percent, would be acceptable.

            Well, a decade later, in 1998, based on industry request, in addition to the mean of greater than equals 75 percent, FDA added a division for less than--or equal 9 percent, to eliminate some of the outliers.

            In 2004, when FDA reviewed some of the new submissions, we noticed that some of the studies, they can meet mean and the standard division.  However, the number of individual units that has recovery less than 75 percent is high.

            For example, five out of 20 units, or six out of 20 units has individual recoveries less than 75 percent, and this raised the concern about the quality of the RBC products, and to ensure the proportional successes, another parameter which is proposal for units with recovery greater than equals 75 percent was a one-sided 95 percent lower limit, greater than 70 percent, was added to the mean and standard division.

            So this was also called the current exemption criteria, and this criteria was discussed at 2004 BPAC meeting, and it was also communicated with the regulated industries through pre-meetings, and it was also presented at a couple of different workshops.

            So since 2004, majority of the submissions to FDA in this time period passed the current acceptance criteria.  Those that do not meet the current acceptance criteria also failed the previous mean and standard division criteria.

            So we believe that maintenance of quality for new RBC products is important.  Reduction in the approval of clearance criteria would allow RBCs to be the more severe damage, storage lesion to the market, which may correlate with a poorer clinical outcome.

            In next couple slides, I'm going to show you example for some retrospective analysis for transfusion of aged red blood cells associated with the adverse clinical outcome.

            This paper, published in 2006, titled as the Association  Between Duration of Storage of Transfused Red Blood cells and Morbidity/Mortality After Reoperative Cardiac Surgery.

            This shows that when you transfuse, the older the blood cells, it is associated with increased in-hospital mortality and associated with increased acute renal dysfunction.

            So the results indicate that there's an association between prolonged RBC storage and adverse clinical outcomes such as mortality and organ failure.

            This paper, published a few weeks ago, talks about the duration of red cell storage and the complications after cardiac surgery.

            The X axis shows the year the patient received the red cell transfusion, and the Y axis shows the survival.

            The orange line shows the patient received the new red blood cells which is less than 14 day old, and the blue line shows the patient received older red blood cells, older than 14 day old.  And the study shows that the patient has a better and higher survival when they receive the newer red blood cells.

            So here are the reasons to revise the in vivo RBC recovery and acceptance criteria.  For example, we have three studies, studies A, B, and C, and the sample size, 24, 21, 21, and all three studies met the acceptable criteria for mean recovery greater than 75 percent, and all three studies met the standard division of less than or equal 9 percent.

            However, in study A, there were eight individual units, has RBC recovery less than 75 percent, that is, eight out of twenty-four, and that makes the one study, 95 percent lower cost B- a limit for proportion of units having recovery greater than equals 75 percent, was only 47.9 percent.

            And in study B, we have five out of 21 units has RBC recovery less than 75 percent, and that makes the proportion of units having recovery greater than 75 percent, 56.3 percent.

            Study C, two out of 21 failures, and that makes the proportion of units having recovery greater than 75 percent was 72.9 percent.

            So the products from the study C has a much higher proportion of units having recoveries greater than 75 percent.

            So the high rate of individual units is that it's A and B, raised a concern about the quality of RBC products, and led FDA to consider revising the criteria.

            This slide shows, further explains the three studies that we mentioned in the previous slide.  The red line shows the RBC 24-hour recovery at 75 percent, and was again that all three studies met, the mean met the greater than 75 percent, and all three studies had the standard deviation, less than 9 percent.

            However, the number of individual units that has recovery less than 75 percent was that eight out of 24 from study A, and five out of 21 in study B, and only two out of 21 in study A.  Therefor, the products from study B offers much higher proportion of the units has individual recovery greater than 75 percent.

            So the revised current acceptance criteria emphasize the population proportion of successes, also called the 95-70 rule.  It is to ensure that most products, meaning greater than 70 percent of the products, have recovery greater than equals 75 percent.

            To meet this 95-70 rule, a specific number of maximum failures are allowed in a study, depending on the sample size of the study.

            For example, if the sample size of the study is twenty, then the number of units with recovery less than 75 percent would be two, and three out of 24, four out of 28, five out of 33.  So if the clinical study met this kind of outcome, one can conclude that with one study, 95 percent lower count limit, greater than 70 percent of the units will have RBC recoveries greater than 75 percent.

            So here comes the key issue we are going to discuss today.  Should the RBC in vivo recovery threshold value of greater than equals 75 percent be changed?

            The reason we ask this question was that since 2004, after FDA institute the revised criterion, some of the manufacturers raised the concern that some of the new RBC products may not be able to meet the current acceptable criteria, and the products already on the market would not meet the criteria either.

            Therefore, the manufacturers volunteered to provide the RBC in vivo recovery study data used for the support and approval of RBC products already on the market, to reassess the current criterion.

            And Drs. Dumont and AuBuchon collected the data and analyzed the data.  The objective of the BEST study was to determine if the products already on the market would be able to meet FDA's current acceptance criteria, and the BEST study concluded that the overall data analysis from 1990 to 2006 will not meet FDA's current acceptance criteria unless the threshold value changed from 75 percent to 67 percent. 

            Well, FDA also analyzed the data after Dr. Dumont kindly shared the BEST data with FDA and here's the FDA analysis of a combined BEST and FDA dataset in different time period, from 1990 to 2007.

            Well, this is a very complex table that Dr. Kim is going to go through this table in great detail in her talk later.  I'm only going to point out a few key points from this table.

            First of all, the BEST data collected three subsets of the studies.  One is the conventional 42 days liquid stored with the blood cells.  Another set, the gamma-irradiated red blood cells.  Another set is the frozen blood cells.  Today, we're only going to focus on the 42 day conventional liquid stored red blood cells to assess the current acceptance criteria.

            The gamma-irradiated red blood cells and the frozen red blood cells are considered as the special circumstances and they will be discussed at other times.

            Second is that here's the--595 are from the BEST datapoint.  So in addition to the 595 BEST datapoint, FDA added an additional 94 datapoint.  That was from the full approvals from the 2004 to 2007, which were not included in the BEST data analysis.

            And so that makes the final N of the 689.  Well, also to point out that studies 40 and 41, that were included in the BEST analysis, were not included here because the information of the year study was not available and we were not able to put into any of this time period.

            Although studies 40 and 41 are excellent studies, both studies met the FDA's current acceptance criteria.  And so we analyzed the--so in contrast to the BEST analysis, which lumps all the datasets from 1990 to 2006, to see if that meets the current acceptance criteria, FDA actually analyzed the data in three time period based on the year of the acceptance criteria use.

            For example, 1990 to 1997, the acceptance criteria was to meet the mean of greater than 75 percent only.  From 1998 to 2003, we added standard division of less than nine.  In 2004 to 2007 added a proportion greater than 70 percent.

            So if you look at the third value of greater than equals 75 percent, the success rate to meet this value increased with time, for example, from .83 to .93, the current time period.

            And the success rate--the power to meet the current acceptance criteria also increased from .43 to .92, and the number of the studies to meet the current criteria increased from the first time period of four out of eight to nine out of eleven, and to eight out of eight.

            I would also like to point out that the success rate, or the datapoints to meet greater than 75 percent is more than 90 percent, if you combine the datapoint from 1998 to 2003, to 2004, 2007, more than 90 percent of the datapoints can meet the threshold value of greater than 75 percent, meaning only 10 percent of the datapoints cannot have the recovery greater than 75 percent.

            So here are the observations of the combined data analysis.  Overall, the quality of RBC products approved or cleared by FDA is improving with time.  Most recent RBC products, meaning from 2004 to 2007, submitted to FDA passed the higher standard with a power of .92, with a threshold value of greater than equals 75 percent.

            This actually answered one of the concerns, that the manufacturer concern that some of the new products are unable to meet the current acceptance criteria, and it is also known that the most clinical studies performed to satisfy FDA criteria for drugs are powered at .80.

            So the threshold value of greater than equals 75 percent has provided a standard for RBC quality evaluation over the last 24 years.  The current criteria show that most of the RBC products, meaning greater than 70 percent, have a recovery greater than equals 75 percent.

            Therefore, based on these considerations, FDA proposes to continue applying the criteria adopted in 2004 to quality evaluation of RBC products using in vivo radiolabeled studies.

            Here are the questions I'm going to ask for us.  I think that we can come back and ask questions again during the discussion.

            So here are the questions to the committee.  Question number one.  Does the committee agree with FDA's proposal to maintain the current criteria? 

            The current criteria are: Radiolabeling studies should be performed in at least two separate centers with a total of 20-24 healthy donors.

            The mean recovery at 24 hours for each unit should be greater than equal to 75 percent with a standard division of less than equal 9 percent; and the one-sided 95 percent lower confidence limit for the population proportion of successes greater than 70 percent.  Here, the success means that each individual of RBC recovery greater than equals 75 percent.

            Question number two.  Alternatively, does the committee recommend that a change in the criteria is needed based on the data presented today?

            And question number three.  If the answer to question two is yes, what changes does the committee recommend for the threshold value of individual subject RBC in vivo recovery with a sample size of 24?

            Examples to consider.  We give you three examples here.

            A.  based on the combined data from '98 to 2007 with greater than equals 74 percent as the threshold value.  Power equals .82.

            B.  Based on combined data from '98 to 2007 with greater than equals 73 percent as the threshold value.  Power equals .93.

            C.  Based on the BEST data from 1990 to 2006, BEST recommends 67 percent as the threshold value.  Power equals .999.

            And I would like to take this opportunity to thank all the FDA staff for their expertise and support, and I also like to thank Dr. Dumont and Dr. Jim AuBuchon for sharing the BEST data with us.  And thank you for listening.

            DR. SIEGAL:  Thank you, Dr. He.

            Questions?

            DR. SZYMANSKI:  Now for clarification, when you say that the requirement are, now, 75 percent mean recovery--correct?

            DR. HE:  Right.  Yes.

            DR. SZYMANSKI:  Now 9 percent standard deviation?

            DR. HE:  Yes.

            DR. SZYMANSKI:  And then you say if you have 20 to 24 subjects done in at least two different institutions, only three should be failures.  Is that what you say?

            DR. HE:  Yeah.  Three out of 24 meaning--

            DR. SZYMANSKI:  Three out of 24 should be less than 75 percent?

            DR. HE:  Yes.

            DR. SZYMANSKI:  How can you say that you have a mean of 75 percent?  Twenty-four sample studied, and only three is under 75 percent.  Usually, when you have a mean, sort a half is up and the other half is down.  That is like an impossibility.

            DR. FLEMING:  Just to explain, there are two things happening here.  One thing is what is the criterion for success or failure and--

            DR. SZYMANSKI:  She said 75 percent.

            DR. FLEMING:  Right.  So on the individual basis, if your recovery is at least 75 percent, you're called a success, and, in fact, what's being questioned here is should the definition of what, on an individual basis, would be called a success, should it be dropped from 75 to 74, 73, etcetera?  That's the definition of success.

            Now take 20 to 24 people and compute the average success rate, and that average success rate has to be high enough to rule out that it could be 70 percent or lower, and that's achieved by having in 24 people and no more than three failures. 

            So what they're saying is in fact logically consistent.

            DR. SZYMANSKI:  I don't believe so, because if you have 24--no--now if you have 24 people, and only three can be under 75, your mean can't be 75.  Your mean must be over 75.

            DR. FLEMING:  So let me change the scenario, because this type of argument could happen in any disease setting.

            Suppose you have an HIV/AIDS patient and you're trying to reduce viral load.  What do we define success to be?  Maybe we define it to be getting to undetectable levels below 50 cells.  Okay.  Then we define success/failure based on whether a patient has rendered less than 50 cells.

            Now take 24 such patients, and can we ensure the success rate in those 24 patients is at least above 70 percent?  That's the same situation here.

            So what's confusing, somewhat, is what's defined to be success is a 75 percent recovery.  That's the individual basis for defining success.  Now take 24 people and conclude that the success rate is at least 70 percent, which occurs when you see at least 21 of 24 successes.

            DR. HE:  Right.

            DR. DI BISCEGLIE:  Can I offer a clarification?  I'm not sure how this, how these criteria are used.  I mean, what is being served by the--the blood bank center product?  I don't know how these things are used.  Please clarify.  Well, you said these are acceptance criteria.

            DR. HE:  Right.

            DR. DI BISCEGLIE:  Acceptance of what?

            DR. HE:  Accept the in vivo recovery studies.

            DR. DI BISCEGLIE:  I'm sorry.  This is probably obvious to the hematologists in the room but I have not--

            DR. VOSTAL:  -- new products for isolating, collecting, processing red cells.

            DR. DI BISCEGLIE:  And device?

            DR. VOSTAL:  For a device; yes.  Let's say a storage bank.  You get a new storage bag for evaluation, we want to make sure that the quality of those red cells stored in that bag are assured for 42 days.  So we do a study at the end of 42 days, this radiolabeling study, and we set the criteria, that the recovery should be greater than 75 percent with these additional statistical criteria.

            DR. HE:  Okay; thank you.

            DR. SIEGAL:  Any other questions or discussion?

            DR. DI BISCEGLIE:  If I may.

            DR. HE:  Sure.  Please.

            DR. DI BISCEGLIE:  You show the data on survival versus--survival, patient survival versus age of the blood.  Are there any data that correlate patient survival or clinical trials to red cell viability?

            DR. HE:  Well, no, we don't have data on that, but we do know that when the red blood cells store longer, and then the survival drops.  And we know that, many of the late studies showing that the aged red blood cells also associated with adverse clinical outcomes.

            So here we give the example, wants to try to say that before a device, or before anticoagulant, for storage of the red blood cells to be on the market, we want to assure that the survival is stored in those--the red cells survive, stored in those bags, will be higher than 75 percent.

            DR. DI BISCEGLIE: Is that certain?

            DR. HE:  It is not but, however, I can tell you that the 75 percent recovery has been debated in the past, many, many years.  In fact, since 1947, that was 60 years ago, Dr. Rose already published his paper saying that when you're evaluating a new RBC collection bag, or RBC storage, storage in solution, and you want to ensure that more than 70 percent of the red blood cells stored in that device are viable.  Okay?

            And that was 60 years ago.  At that time the red blood cells were stored in the glass bottles, in a much less advanced anticoagulant such as ACP.  And this is why, 20 years ago, in 1985, the experts in the field, they feel that, well, 40 years ago, in 1947, the experts were ready to propose and to suggest that the recovery should be greater than 75 percent.

            Now, in 1985, we have much more advanced additive solutions, we have plastic bags, and the survival should be increased a little bit higher.  That's why, at that time, they increased recovery, 24 hour recovery to 75 percent as the threshold value.

            And also I'd like to point out that the most recent combined study from FDA and BEST was 689 datapoints.  You can see that.  More than 90 percent of the datapoints from that study, showing that more than 90 percent of the datapoints can meet the 75 percent recovery, meaning only 10 percent of the datapoints will not be able to meet 75 percent recovery.

            DR. ZIMRIN:  You showed two studies that suggested that with older red cells, associated with the adverse clinical outcome--just a point of clarification.  There are certainly studies that do not suggest that.  You didn't mention that in your presentation; right?

            DR. HE:  That is true.  That is definitely true.  But on other hand, the study I showed, the second one, that's probably one of the largest study, and from a single center.  They'd actually be able to separate, to actually differentiate that some people only received the new red blood cells and some people only received the old red blood cells, and the people who received the new red blood cells has a better survival.

            Of course understand, we all understand that there are some variations, and there are some other points, should be further studied and further analyzed, and also this is a retrospective study--

            DR. ZIMRIN:  It's a retrospective study.  The graph you showed was an unadjusted comparison; right?

            DR. HE:  We all understand that debate but--

            DR. ZIMRIN:  Well, no, you did make that point.

            DR. HE:  Right.  We understand that; yes.

            DR. DI BISCEGLIE: I don=t know if this is the point you're trying to make.  I think, you know, we need to have a clear understanding if this is or is not going to be significant, but I'm not sure you've really answered the question.  I think you're saying the question can't be answered.

            DR. GOLDING:  Yes.  You know, I think you are pointing out relevant points, and we agree with what you're saying.  What we're saying is the current state of knowledge is not perfect, but what we do know is that we would--that it's likely, that if you have more viable cells, and you transfuse them, that there's less chance, we think, of getting adverse events.

            You know, it might take another five or ten years before the studies that are being alluded to are done in a more satisfactory way.  Do we want to reduce our standard in the meantime, before those studies are completed, or do we want to maintain a standard that we think allows most of the products that we see to be approved at that level?

            And the tests that we have are not perfect.  So the 75 percent survival, how does that relate to a clinically meaningful outcome? which is your question.  We don't have a definitive answer to that, but our approach is that this test has stood the--has been used as a criterion and can assure a certain viability of red cells, and that we'd like to maintain the criterion unless we have evidence to say that it's not a good approach in terms of approving products, and trying to assure that these products are safe in effect.

            DR. FINNEGAN:  I'm even less sophisticated than the hematologist.  Are you saying that we are discarding good blood?  Or are you saying that you're using this to test new bags and new fluid?

            DR. HE:  This is a test of new bags and new fluid.

            DR. FINNEGAN:  So we're not throwing blood away?

            DR. HE:  No; no.  Not for already approved on the market.  Only for the new bags.  Yes.

            DR. SIEGAL:  Just to clarify for me, these are blood samples that have sat for 42 days.

            DR. HE:  Right.

            DR. SIEGAL:  So they're way at the end of the shelf life.

            DR. HE:  Storage period; yes.

            DR. SIEGAL:  And they have no bearing, really, on the New England Journal article that just came out, really, which tested 15-day-old as the cutoff, where you have a clinical end point, blood.

            DR. GLYNN:  And again, these studies, the two studies you refer to, these are retrospective studies?

            DR. HE:  Right.

            DR. GLYNN:  I don't think there are any--they haven't been--well, there have been one pilot clinical trial done so far on just like 57, 59 patients, but otherwise there are no prospective data that I'm aware of.

            DR. HE:  That's true.  Yes.  Thank you.

            DR. SIEGAL:  Dr. Epstein.

            DR. EPSTEIN:  Yes.  Back to Dr. Zimrin's point which is well-taken.  Two things.  First, the Advisory Committee for Blood Safety and Availability will be addressing the question of what do we really know about age of storage in relation to clinical outcome, and I think we would readily concede that we don't know the answer now because the available studies go both directions, some showing no effect, some showing an effect, and the vast majority are retrospective and they're full of confounders.  So it's a little bit of a distraction.

            The reason for having mentioned that is that there's at least some body of data suggesting that there is a storage lesion.  We didn't have to cite those data.  You can see that, just from the reduced recovery, or the slide that was shown based on the studies done by Mollison, shows that age of storage correlates with reduced viability of the cells you infuse, or you recover a smaller percent the longer you store the blood.

            So we know that there's damage to cells.  So the point here really is we also know that the survival of the infused red cells relates directly to the proportion that are nonviable and are rapidly cleared.

            So what we're basically saying is if you're going to transfuse red cells, what you want is that they should be functional in vivo.  We think that the length of survival correlates with functionality.  In other words, the body's getting rid of bad cells.

            So the test at 24 hours for recovery is actually a surrogate, it's a predictor for the survival of the red cells in vivo, which we think is a surrogate or predictor for their functionality in vivo.

            So what we're basically saying is that it's a quality characteristic, that if the recovery is better, it means you've damaged the cells less, and we think that that has to be good.  So it's not that we're linking it directly to knowledge about the clinical outcome of aged blood versus fresh blood.  It's that we believe that all stored blood has a storage lesion, and that is shown by the fact that recovery goes down and survival goes down the more you store, and all we're saying is, well, if you look at the end of the storage period, we want to set a lower limit on the amount of nonviable cells that are in the population, cause we think that that's a quality factor for the product.

            So, again, the clinical outcomes with age versus fresh blood is a little bit of a distraction.  It was only there to illustrate that we know there's a storage lesion.  But we know that anyway.

            DR. SIEGAL:  Dr. Szymanski.

            DR. SZYMANSKI:  I agree with that but what I have difficulty with is to set these requirements, as presented, without the data that now exist, because it feels to me that here statistics are used to "strangle" the biology, and we know that there are, when you do the survival studies, there is individual variation, and there are certain donors and recipients who are their o