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Biomedical subjects

D H Buchholz

Publications and source records attributed to D H Buchholz.

13 recordsLinked to original sources

Ultraviolet-B irradiation of platelets: a preliminary trial of efficacy.

Prior studies established that ultraviolet-B light (UVB) irradiation of platelet concentrates (PCs) at appropriate doses can eliminate the mixed lymphocyte culture-stimulating and -responding capacity of lymphocytes in the PCs without adversely affecting in vitro platelet function. The in vivo recovery and survival and in vitro characteristics of UVB-irradiated platelets were investigated in paired studies. PCs were stored for 1 day and then exposed to UVB. Platelet recovery, survival, and function were comparable to those of nonirradiated platelets. Recovery and survival of platelets stored for 5 days before UVB exposure were decreased relative to controls, although they were considered clinically acceptable. Paired transfusion studies were also performed in seven thrombocytopenic patients by using platelets obtained by apheresis. Comparable posttransfusion platelet increments and bleeding time corrections were obtained with both irradiated and control (nonirradiated) platelets. It can be concluded that platelets survive and function relatively normally in vivo after UVB irradiation sufficient to abolish lymphocyte reactivity in mixed lymphocyte culture. Long-term studies of UVB-irradiated PCs are needed to assess their potential in reducing recipient alloimmunization.

Blood Component Transfusion

Removal of Yersinia enterocolitica from AS-1 red cells.

The growth of Yersinia enterocolitica in AS-1 red cells was investigated so as to study the organism's proliferation kinetics and to evaluate the effect of prestorage white cell (WBC) reduction on bacterial multiplication. Twenty-four 2-unit pools of ABO-compatible whole blood were prepared and inoculated with Y. enterocolitica to final concentrations ranging from 0.3 to 132 organisms per mL. After inoculation, pools were split equally, AS-1 red cells were prepared, and 1 unit of each pair (test unit) was WBC-reduced with a WBC-reduction filter. Quantitative bacterial cultures of both WBC-reduced and control units were performed at several points throughout preparation and storage. Less than 10 percent of the inoculated organisms was recovered from blood samples taken after a 7-hour room-temperature holding period. By the end of 42 days of storage, Y. enterocolitica was recovered from unfiltered red cells in 2 of the 6 units inoculated at the lowest levels (0.3 and 0.7 organism/mL), from 8 of the 12 units inoculated at the intermediate levels (2.8, 5.2, 30.7, and 43 organisms/mL) and from 6 of the 6 units inoculated at the highest levels (98.8 and 132 organisms/mL). Positive cultures were seen as early as Day 7. In contrast, filtered units inoculated at all levels less than or equal to 98.8 organisms per mL (21/21 units) were sterile at the end of the 42-day storage period, while 2 units (2/3) inoculated at 132 organisms per mL showed growth despite filtration.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Five-day storage of platelets in a non-diethylhexyl phthalate-plasticized container.

A non-diethylhexyl phthalate (DEHP)-plasticized blood bag for 5-day storage of random-donor platelet concentrates has been developed. The plastic bag is composed of polyvinylchloride plastic with a butyryl trihexyl citrate plasticizer. The suitability of this plastic for the storage of platelet concentrates for use in clinical transfusion practice was evaluated. In vitro storage studies showed no significant differences at Day 5 for a series of in vitro assays (test plastic vs. control plastic) including pH (7.31 vs. 7.44), lactate dehydrogenase discharge (21.8 vs. 17.1%), pO2 (103 vs. 120 torr), osmotic recovery (52 vs. 57%), and morphology score (527 vs. 516). For paired radiolabeled recovery and survival data from autologous blood donors, results showed equivalence between the test plastic and two control plastics. A small but significant difference between test and control plastics in regard to survival was found by using a linear computer model, but not with a gamma function (multiple-hit) model. For paired transfusions to thrombocytopenic patients, the corrected count increments at 1 to 4 hours (test vs. control) were 13,534 versus 15,494 (p > 0.05, NS). Similar results were seen for corrected count increments determined at 12 to 24 hours. It can be concluded that platelets stored in the test plastic are acceptable for use in clinical practice.

Blood Platelets

Long-term granulocyte transfusion in patients with malignant neoplasms.

Thirty-eight episodes of culture-documented antibiotic-resistant bacterial or fungal infection in patients with malignant neoplasms were treated with daily granulocyte transfusions until the infection improved or the patient died. Cumulative summation temperature plotting allowed easier interpretation of recipient fever response. Seventy-one percent of recipients had a favorable response to transfusion. There was no difference in mortality between patients treated with cells collected by filtration (FL) or intermittent flow centrifugation (IFCL) leukapheresis techniques. Transfusion reactions were more than twice as common with FL than IFCL collected cells. Seventy-four percent of recipients were alive 21 days after completion of transfusions; of the ten deaths, five could be classified as granulocyte transfusion failures. This study suggests that long-term granulocyte transfusion may be required in infected recipients when autologous granulocytes do not return after chemotherapy.

Abscess

Clinical experience with transfusion of granulocytes obtained by continuous flow filtration leukopheresis.

Granulocytes obtained by continuous flow filtration leukopheresis (CFFL) were transfused to 21 patients on 131 occasions. An average of 28.2 times 10-9 granulocytes were administered per transfusion. These cells were more than 90 per cent viable by dye exclusion, ingested latex particles normally and had almost normal bactericidal activity. Migration to skin windows was demonstrated on four of six attempts, but 51-Cr-labeling studies failed to show localization in infected areas on six occasions. Post-transfusion granulocyte count increments averaged 225/mul and were transient. Significant transfusion reactions occurred during 35 transfusions to 13 patients. Reactions occurred in some patients without demonstrable alloimmunization and after six infusions of HL-A identical or compatible cells. Definite clinical improvement was noted in three recipients. Stabilization of infection with patient survival occurred nine times, and progression of infection with death eight times. Granulocytes obtained by CFFL are viable and functional. Their transfusion is not without risk and must still be considered an investigative procedure of suggestive but as yet unproved clinical efficacy.

Acute Disease

Granulocyte harvest for transfusion: donor response to repeated leukapheresis.

Leukapheresis of normal donors with the NCI-IBM Continuous Flow Blood Cell Separator was compared with the method of filtration leukapheresis. An average of 5.7 times 10-9 (range 1.4 to 10.9 times 10-9) granulocytes were collected on 25 occasions with the blood cell separator compared with an average of 36.6 times 10-9 (range 20.1 to 61.3 times 10-9) obtained by filtration leukapheresis on 85 occasions. Donor platelet counts decreased following donation by an average of 29,000/mul (14%) with continuous flow centrifugation (CFC) and an average of 40,000/mul (17%) with filtration leukapheresis (FL). Estimated donor packed red blood cell loss, including blood obtained for investigative purposes, was 80 ml per donation with CFC and 67 ml per donation when FL was used. FL resulted in at least a 25 per cent increase in the granulocyte count of donors by donation completion in 61 per cent of donors, although postdonation granulocytosis did not correlate with increased cell harvest. CFC donors generally displayed a postdonation decrease in granulocyte count. Normal numbers of granulocytes were maintained in donors undergoing frequent leukapheresis and no donor's health was compromised by the removal of large numbers of granulocytes. Repeated donations were possible with both systems. Although platelet counts dropped more with FL, the procedure was performed as frequently as eight times in a ten-day period without development of severe donor thrombocytopenia. Donor red blood cell loss appears to be the limiting factor in the repeated use of either system. Blood loss could be significantly decreased under routine conditions if fewer samples were obtained for investigational purposes. Filtration leukapheresis offers the advantage of significantly greater granulocyte yields than is possible with CFC unless modifications such as arterio-venous shunts or the use of steroids and/or rouleaux-inducing agents are employed.

ABO Blood-Group System

Unusual response to ABO incompatible blood transfusion.

Three units of group A blood were inadvertently administered to a group O recipient during surgery without evidence of hemoglobinemia, hemoglobinuria, hypotension, disseminated intravascular coagulation, acute renal tubular necrosis, or other signs and symptoms of transfusion reaction. The recipient had normal concentrations of IgG, IgA, and IgM as well as complement (C3) prior to transfusion and anti-A agglutinins titered to 64 (titer of 128 by the antiglobulin technic). Seventeen hours following the transfusion, 28 per cent of the circulating red blood cells were group A (equivalent to 475 ml of packed cells); they were eliminated by day 5 without evidence of hemoglobinuria, hemoglobinemia or hyperbilirubinemia. Anti-A titers (antiglobulin) had risen from a posttransfusion low of 4 to 4,096 by day 10. After treatment of serum with 2-mercaptoethanol, however, hemolytic activity which was first noted on day 5 was lost and the antiglobulin titer dropped to 24 which suggested that most of the anti-A produced in response to the transfusion was IgM rather than IgG. The anti-A titer had dropped to essentialyy pretransfusion levels and the majority of anti-A present was IgM by day 91. The recipient suffered no untoward effects from the transfusion and was in good health three months following the transfusion.

ABO Blood-Group System

Frozen autologous platelets in the supportive care of patients with leukemia.

Multiple units of platelet concentrate obtained by intensive plateletpheresis of patients with leukemia in remission were pooled and frozen using 4 to 5 per cent dimethylsulfoxide and retransfused during periods of thrombocytopenia. Plateletpheresis was well tolerated by all donors and an average platelet yield per unit of 0.99 X 10(11) (n = 155) was obtained. The results of 107 transfusions to 36 patients are presented. An average of 32.4 per cent of the platelets were lost during the freeze-thaw process. Freezing loss was lowest at a freezing rate of one degree C per minute, at a lower final concentration of platelets, and when polyolefin bags were used. The mean corrected posttransfusion count increment was 6,400/mul (range 600-19,000 xm2/10(11) platelets transfused). In vivo results did not correlate with freezing rate but were statistically significantly better at lower platelet (approximately 0.16 X 10(11) platelets/10 ml) concentrations. Eleven patients, including some who were refractory to random donor platelets were supported entirely with autologous platelets during reinduction therapy for leukemia. When administered prophylactically the autologous platelets seemed to prevent hemorrhage during periods of thromobocytopenia although in most patients bleeding times were not corrected posttransfusion. This study demonstrates that frozen autologous platelets can be used in the supportive care of thrombocytopenic patients. Further technical improvements are necessary before platelet freezing becomes practical for widespread use.

Blood Cell Count

Plasmapheresis using the IBM 2991 blood cell processor.

The IBM 2991 Blood Cell Processor, normally used to wash or deglycerolize red blood cells, has been modified to permit serial plasmapheresis using a small roller pump, an adapter line and the disposable processing set usually used for cell washing. Blood is withdrawn from the donor, pumped into the plastic centrifuge bag and the plasma removed after centrifugation. Donor erythrocytes are pumped from the processing bag to a reservoir bag from which they are returned to the donor by gravity. Multi-unit plasmapheresis to harvest plasma for transfusion or to remove plasma as a therapeutic measure is possible using the modified system. A four-unit plasmapheresis can be done in 65 to 75 minutes at a cost of $27.61, including disposables and technologist labor. This modification allows the IBM 2991 to function for red blood cell washing and as an efficient instrument for plasmapheresis.

Blood Donors

Storage of platelet concentrates after high-dose ultraviolet B irradiation.

Ultraviolet B (UVB) irradiation of platelet concentrates (PCs) may prevent the development of posttransfusion HLA alloimmunization. As irradiation performed in a blood center or a hospital will probably be associated with a variable postirradiation delay before transfusion, the ability to store PCs after UVB irradiation becomes important. The effects have been studied of a UVB dose of 10,000 mJ per cm2, the dose used in our institution for UVB clinical trials, on PCs pooled and stored for up to 96 hours after irradiation. Results showed that after 96 hours of storage, though there were no changes in pH, platelet count, white cell count, percent discharge of lactate dehydrogenase, or beta-thromboglobulin, there were significant decreases in morphology score and osmotic recovery. These changes, however, were not evident after 24 hours of storage. Similarly, there was a 60-percent decrease in immunoreactive membrane glycoprotein (GP) Ib after 96 hours of storage, but these changes were not seen after 48 hours of storage. No changes were seen in levels of GPIIb/IIIa in either group during the 96 hours of storage. On computer-analyzed two-dimensional polyacrylamide gel electrophoresis, PCs irradiated at 10,000 mJ per cm2 and stored for 72 hours had changes in over 50 platelet proteins as compared to those proteins in nonirradiated age-matched control PCs. It can be concluded that UVB irradiation of PCs at 10,000 mJ per cm2 does not lead to significant platelet deterioration after short-term storage (24-48 hours) but is likely to be deleterious after long-term (72-96 hours) storage.(ABSTRACT TRUNCATED AT 250 WORDS)

Blood Banks