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S Holme

Publications and source records attributed to S Holme.

At least 37 records · Page 2Linked to original sources
PubMed1995

In vitro platelet ageing at 22 degrees C is reduced compared to in vivo ageing at 37 degrees C.

In these studies, platelet ageing during in vitro at 22 degrees C was compared with in vivo ageing using isotope labelling. Paired fresh and 5-d-stored platelets had a mean residual life-span (MRL) of 4.8 +/- 0.7 d and 3.2 +/- 0.9 d, respectively. After 2.1 +/- 0.4 d in vivo circulation, the MRL of the fresh platelets was equivalent to that of the 5-d-stored in vitro platelets. This suggests that platelet ageing for 5 d in vitro at 22 degrees C corresponds to 2.1 d in vivo ageing at 37 degrees C. Thus, the relative ageing at 22 degrees C in vitro was (2.1 d/5 d) = 0.42 of that at 37 degrees C in vivo. A similar ageing ratio (0.44) was obtained by measurement of the decrease in MRL during storage at 22 degrees C of platelets stored for 1, 5, 7, 10 and 14 d relative to the decrease in MRL of fresh platelets in vivo. ATP turnover rate at 22 degrees C was compared to the rate of 37 degrees C by measurement of the rates of platelet oxygen consumption and lactate production in vitro. In vitro ATP turnover at 22 degrees C versus 37 degrees C, was found to be 10.5 +/- 1.0 versus 21.6 +/- 1.4 mumol/10(12) plts/min, respectively. Thus, the ATP turnover ratio (0.48) at these two temperatures suggests that the relative decrease in ageing at 22 degrees C compared to 37 degrees C is similar to the relative decrease in metabolic rate at this temperature.

Adenosine Triphosphate↗
PubMed1994

Sustained elevation of intracellular cyclic 3'-5' adenosine monophosphate is necessary for preservation of platelet integrity during long-term storage at 22 degrees C.

Preservation of platelet integrity and responsiveness was examined in platelet concentrates prepared in the presence of various formulations and combinations of platelet-activation inhibitors affecting intracellular levels of cyclic 3'-5' adenosine monophosphate (cAMP). Platelet concentrates were prepared and stored in an artificial medium for two weeks at 22 degrees C. Markers of metabolic activity (pH, lactate, pO2, pCO2 in the medium), aggregation response, hypotonic shock response, and glycoprotein Ib (GPIb) expression were assessed along with direct measurements of cAMP in platelet pellets and thromboxane B2 (TxB2) in the supernate. The platelet concentrates prepared with only adenylate-cyclase stimulators (prostaglandin E-1 or forskolin) showed less maintenance of the integrity and responsiveness markers and greater loss of GPIb than concentrates prepared with phosphodiesterase inhibitors (theophylline or caffeine) or combinations with the above. These results were correlated with the ability of these compounds to sustain elevation of cAMP above basal level during the entire extended-storage period. The strong correlation (rs = -0.67) between elevation of cAMP levels and suppression of TxB2 production suggests that the phosphodiesterase inhibitors provided better protection than stimulators of adenylate cyclase alone through a reduction in platelet activation and its deleterious effects on preservation of platelets during storage.

Blood Platelets↗
PubMed1994

Characteristics of white cell-reduced red cells stored in tri-(2-ethylhexyl)trimellitate plastic.

BACKGROUND: Standard blood storage containers contain extractable plasticizers that accumulate in blood during storage and are an unintended transfusion product. However, extractable plasticizers have a protective effect on the red cell membrane and improve red cell storage variables. Prestorage white cell reduction also improves selected red cell storage variables. STUDY DESIGN AND METHODS: The study evaluated whether the beneficial effect of prestorage white cell reduction would offset the negative effect of the absence of extractable plasticizer in red cells stored in AS-3 for 42 days at 4 degrees C. Filtered red cells stored in polyvinylchloride containers with the nonextracting plasticizer, tri-(2-ethylhexyl)trimellitate (TEHTM), were compared to unfiltered red cells stored in polyvinylchloride containers with the extractable plasticizer di-(2-ethylhexyl)phthalate (DEHP). RESULTS: Poststorage supernatant potassium and red cell osmotic fragility were significantly higher in white cell-reduced TEHTM units than in unfiltered DEHP units. The mean 24-hour recovery of the filtered TEHTM red cells was significantly lower than that of the unfiltered DEHP red cells (69.1 +/- 7.4% vs. 77.1 +/- 5.1%, p < 0.05, n = 8). CONCLUSION: These data demonstrate that white cell reduction before 42-day storage in TEHTM containers with currently approved preservatives does not yield an acceptable red cell component.

Adenosine Triphosphate↗
PubMed1994

Evaluation of platelet concentrates stored for 5 days with reduced plasma volume.

BACKGROUND: Currently, platelet concentrates (PCs) are stored in a suspending plasma volume of 45 to 65 mL. Previous studies using second-generation containers indicated that PCs stored for 5 days at volumes less than 30 mL have reduced in vivo percentage recoveries as compared to PCs stored at volumes of 50 mL or more. STUDY DESIGN AND METHODS: This study has evaluated the effect of PC plasma volume on the maintenance of in vivo and in vitro platelet properties following 5 days of storage, with the purpose of establishing the minimum plasma volume in the range of 30 to 50 mL. Twenty paired studies were performed in which identical populations of platelets from the same donor (obtained by double manual apheresis) were stored in a normal volume (55-60 mL, control) and reduced volume (30-50 mL, test) of plasma. Comparison of in vivo viability between test and control PCs was performed after random radiolabeling of 1 unit with 51Cr and of the other with 111In, with simultaneous transfusion and with calculation of percentage recovery and the area below the survival curve (integral) as measures of viability. RESULTS: When test unit volumes were > or = 35 mL, essentially identical platelet survival curves and in vitro results were obtained for test and control. The integral and the percentage recovery for the test units were (mean, 95% confidence interval) 98.7 (96.3-101.0) and 99.0 percent (94.7-103.3) of those values in the control units, respectively. Test units with volume < or = 34 mL demonstrated reduced in vivo viability with integral and percentage recovery of 81.1 (68.9-93.3) and 80.4 (69.3-91.5), respectively, as compared to the control units. This loss was associated with increased metabolic activity (lactate production), which may suggest platelet activation due to the increased surface-to-PC volume ratio. CONCLUSION: These results show that the storage volume of PCs may be reduced from 50 to 60 mL to 35 to 40 mL without any significant decrease in in vivo or in vitro platelet quality.

Blood Platelets↗
PubMed1994

Effect on platelet properties of exposure to temperatures below 20 degrees C for short periods during storage at 20 to 24 degrees C.

BACKGROUND: When platelet concentrates (PCs) are shipped over long distances, it is not always possible to ensure that their temperature is maintained at 20 to 24 degrees C. In addition, PCs are not agitated as during routine storage. STUDY DESIGN AND METHODS: Studies have been conducted to evaluate how exposure to temperatures below 20 degrees C in the absence of agitation influences properties of platelets. In initial studies, exposure to 4 degrees C for 3 or 5 hours or to 12 degrees C for 5 or 17 hours on Day 2 of a 5- to 6-day storage period was associated with a loss of discoid shape. This was reflected by slightly lower but statistically different morphology scores after storage compared to those observed with control platelets that were stored only at 20 to 24 degrees C. In addition, a qualitative difference in morphology was noted in controls and PCs held at 16 degrees C for 17 hours. In more detailed studies, both the in vivo viability and in vitro properties of platelets exposed between Day 1 and Day 2 to either 12 degrees C or 16 degrees C for 17 hours were evaluated. The protocol involved a paired study design (n = 4 for each exposure temperature) with the simultaneous storage of two identical PCs, one exposed to 12 or 16 degrees C and the other one maintained at 20 to 24 degrees C throughout the 5-day storage. RESULTS: Exposure to 12 degrees C significantly reduced (p < 0.05 by paired t test) the in vivo recovery to 37.6 +/- 13.8 percent (mean +/- 1 SD) from 47.8 +/- 11.5 percent and the survival time to 2.0 +/- 0.3 days from 6.5 +/- 1.4 days. On exposure to 16 degrees C, the differences in viability from those of control units were much less but still significant. The in vivo recovery was 42.7 +/- 3.8 percent compared to 49.2 +/- 3.0 percent and the survival time was 3.5 +/- 1.2 days compared to 6.6 +/- 0.3 days. The loss of in vivo viability of the test platelets was associated with a loss of discoid shape, as reflected by morphology scores, extent of shape change, and mean platelet volume. In addition, platelet metabolism also appeared to be affected, as suggested by increased lactate production. All of the in vitro properties except for total ATP and residual glucose that were statistically different from those of controls on exposure to 12 degrees C were also significantly different on exposure to 16 degrees C. CONCLUSION: These findings demonstrate that platelets undergo substantial changes in in vivo viability and in vitro properties when they are exposed to temperatures below 20 degrees C for short periods.

Blood Platelets↗
PubMed1994

Storage of apheresis platelets after gamma radiation.

BACKGROUND: There are conflicting data on the effect of irradiation and subsequent storage on the quality of platelet components. STUDY DESIGN AND METHODS: The retention of platelet properties during storage of gamma-irradiated apheresis suspensions was studied in 22 apheresis components obtained on a cell separator with a specialized centrifugation chamber. Immediately after collection, each suspension was divided equally into two 1-L polyolefin containers. On Day 1 (n = 12) and Day 3 (n = 10) one of each pair of suspension containers was gamma radiated with 2500 cGy. All platelet suspensions were stored for 5 days at 20 to 24 degrees C. Samples were drawn on Day 5 from each of the 22 pairs of containers for evaluation of an array of in vitro properties. Samples were taken from 10 pairs of containers for platelet labeling with either 51Cr or 111In for subsequent transfusion and concurrent in vivo measurement of recovery and survival. Posttransfusion samples were drawn after 24 hours for ex vivo whole blood aggregation. RESULTS: Comparable in vitro and in vivo properties were measured in irradiated and control platelets, whether irradiation was performed on Day 1 or Day 3. The mean +/- 1 SD in vivo recovery and survival time for controls and platelets irradiated on Day 1 was 52 +/- 14 percent and 146 +/- 34 hours and 51 +/- 7 percent and 147 +/- 36 hours, respectively. For Day 3 irradiation, the values were 46 +/- 12 percent and 150 +/- 60 hours and 47 +/- 9 percent and 151 +/- 53 hours, respectively. A small, but measurable adverse effect of irradiation on ex vivo platelet aggregation was present. CONCLUSION: These data indicate that storage of apheresis platelets after gamma radiation is without clinically significant, demonstrably adverse effects on platelet quality.

Blood Platelets↗
PubMed1994

Effects of 3-5 log10 pre-storage leucocyte depletion on red cell storage and metabolism.

A new, in-line high-efficiency 3-5 log10 leucodepletion filter system (Leukotrap RC system) was used to investigate the effect of pre-storage white cell removal on the quality of AS-3 red cell concentrates stored for 42 d at 4 degrees C. Median residual white cell content was 4 x 10(5) when filtration was performed at 22 degrees C within 8 h of phlebotomy (n = 20) and 3.2 x 10(4) when filtration was performed at 4 degrees C 12-24 h after phlebotomy (n = 24). None exceeded 1 x 10(6) WBC per red cell product. Filtration was rapid (median 28 min), and red cell loss averaged (mean +/- 1 SD) 6.4 +/- 0.7%. In a paired study design, post-transfusion recoveries of 42 d stored red cells in the filtered units averaged 84 +/- 6% v 82 +/- 8% for unfiltered units (P < 0.05) and post-storage haemolysis. ATP, osmotic fragility, K+ and pH were significantly (P < 0.05) better in the filtered units. Reduced glycolytic activity was also observed in the filtered units, and there was a correlation between osmotic fragility, glucose consumption, and lactate produced in standard units that was not present in leucodepleted units. In conclusion, this study suggests that leucodepletion of AS-3 red cell concentrates prior to storage results in better maintenance of the integrity of the red cell membrane with reduced glycolytic activity. There was a modest improvement in post-infusion viability sufficient to offset the filtration-induced loss and to result in an equivalent red cell product.

Adenosine Triphosphate↗
PubMed1994

Evaluation of apheresis platelet concentrates collected with a reduced (30-ml) collection chamber with resuspension and storage in a synthetic medium.

Recently, the CS-3000 Plus Blood Cell Separator with the TNX-6 platelet separation chamber insert has been furnished with a small-volume (30-ml) collection chamber. In this study, a platelet synthetic medium containing glucose and bicarbonate (PSM) was used for resuspension and storage of this highly concentrated platelet product. Eighteen donors participated in a paired study design where each participant donated platelets on two occasions, once following collection in a standard chamber with resuspension and storage in plasma and once following collection in the new chamber with resuspension and storage in PSM. Substantially higher total platelet counts were obtained using platelets collected in the small chamber and stored in PSM as compared to control (4.4 +/- 0.9 x 10(11) vs. 3.5 +/- 0.9 x 10(11) platelets, p < 0.01 by paired t test). After 5 days of storage, PSM-stored platelets demonstrated higher ATP levels, less lactate dehydrogenase in the supernatant and increased lactate production with resulting lower pH at day 5 of storage (6.94 +/- 0.15 vs. 7.08 +/- 0.09, p < 0.05). There were no statistically significant differences of the survival by multiple-hit estimation of PSM-stored as compared to plasma-stored platelets as determined by 111In labeling and infusion. A slight decrease in the initial percent recovery with the additive-suspended as compared to suspended plasma cells was noted: 50 +/- 8 versus 54 +/- 9%, respectively (p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1994

Membrane glycoproteins in cryopreserved platelets.

Although platelets stored by cryopreservation are effective in hemostasis, they acquire a number of functional defects during storage and preparation for transfusion. In addition to known acquired defects such as defective aggregation, decreased resistance to hypotonic shock, and disc-spherocyte transformation, we have shown that cryopreserved platelets have decreased capacity to adhere to subendothelium, compared to liquid-stored platelets. To investigate this decrease in adhesive capacity of cryopreserved platelets, we measured the major adhesive membrane glycoprotein, GPIb, and the principal aggregatory protein, GPIIb/IIIa, using flow cytometry in fresh platelets or in platelets cryopreserved in 5% DMSO. We also analyzed aggregation of cryopreserved platelets or liquid-stored platelets in response to ristocetin as another measurement of GPIb functional capacity. We found that approximately 15% of cryopreserved platelets lost surface-bound GPIb, while there was no measurable loss of GPIIB/IIIa during cryopreservation. The cryopreserved platelets also showed a significant decrease in aggregation to ristocetin, but no loss of response to the stronger agonist, thrombin. The loss of surface GPIb from cryopreserved platelets was modest in degree, approximately that reported for liquid-stored platelets, and does not seem great enough to account for the observed functional changes in aggregation and adhesion.

Blood Platelets↗
PubMed1993

Storage of pools of six and eight platelet concentrates.

Platelet concentrates (PCs) prepared from units of whole blood are routinely stored singly at 20 to 24 degrees C and pooled prior to transfusion. Studies have been conducted to evaluate the in vitro properties of pools of six (n = 19) and eight (n = 17) ABO-identical PCs after storage, with comparative studies involving single units (n = 33). The pools were prepared using the sterile connecting device. One-day-old and 3-day-old PCs were pooled and stored for a total of 5 days in a container system consisting of two 1000-mL polyolefin containers. The pooled platelet suspension was divided approximately equally between the two containers. The platelet count was reduced by less than 5 percent during storage of the pools, which is similar to the reduction found with storage of control units of single PCs. The volume loss due to pooling was 9.6 +/- 1.9 percent (mean +/- 1 SD). The pH of the PC pools was approximately 7.0 after 5 days of storage, with no pool having a pH below 6.2. In vitro platelet properties, such as morphology score, extent of shape change induced by ADP, total ATP, aggregation response to ADP and collagen, response to hypotonic stress, lactate dehydrogenase discharge, and beta-thromboglobulin release, were similar for pools and control single PCs. In addition, comparable low levels of thymidine uptake were detected in the mononuclear leukocyte fraction of pooled and unpooled PCs that were stored for 5 days at 20 to 24 degrees C, which indicates that the mixing of lymphocytes in the pool did not stimulate in vitro immunologic reactions.(ABSTRACT TRUNCATED AT 250 WORDS)

Blood Platelets↗
PubMed1993

Concurrent label method with 111In and 51Cr allows accurate evaluation of platelet viability of stored platelet concentrates.

The precision and reproducibility of 111In and 51Cr platelet radiolabel agents for in vivo kinetic studies of stored platelet concentrates (PC) were investigated. The objective was to develop a precise method with concurrent labelling of two platelet populations using different isotopes, which would allow identification of small differences in in vivo platelet quality. Identical labelling procedures were used to investigate the effects of PC storage age, different methods of red cell (RBC) and white cell (WBC) contamination correction, and label elution correction on the results of 111In and 51Cr kinetic studies. 111In and 51Cr platelet survival curves from the same PC, even when uncorrected for elution and RBC contamination, exhibited excellent correlation, irrespective of the age of the concentrate and its viability. However, slightly higher, but statistically significant, post-infusion per cent recoveries with 51Cr labelled platelets were found. Two factors were identified as the cause for this difference. There was a higher affinity of contaminating RBC/WBC in PC for 51Cr than for 111In. With determination of RBC/WBC activity by centrifugation/density separation, RBC/WBC fractions from the injectate were found to contain 12.6 +/- 3.8% v 7.1 +/- 3.6% of total 51Cr and 111In activity, respectively, in 20 studies. In addition, there was a significantly higher 111In activity in plasma immediately post-infusion than with 51Cr, 5.2 +/- 1.3% v 2.8 +/- 1.6%, respectively, suggesting more label elution or carryover. After correction for the activity of RBC/WBC and for elution or carryover, essentially identical 51Cr/111In platelet survival curves were found. In 31 stored PC studies, the absolute average difference between 51Cr and 111In per cent recoveries was only 4 +/- 3% in a group of donors whose platelet recoveries ranged from 10% to 80%. Similarly, the average difference between 51Cr and 111In survival was only 8 +/- 4 h within a range of survivals from 40 to 220 h. In conclusion, after correction for elution and contaminating RBC/WBC binding, these studies show that 51Cr and 111In may be used interchangeably for labelling of stored PC, and that small differences between test and control platelets could be reliably detected using concurrent labelling with simultaneous infusion.

Blood Component Transfusion↗
PubMed1992

Platelet microvesicles adhere to subendothelium and promote adhesion of platelets.

A role in hemostasis has been suggested for platelet membrane microvesicles (mv). The objectives of the studies reported here include functional analysis of platelet mv in models developed for study of platelet adhesion, as well as investigation of possible interactions between mv and intact platelets in these same adhesion models. Microvesicles were prepared from washed platelet concentrates by repeated freezing and thawing. Adhesion to subendothelium was measured quantitatively by radiolabelling mv with 111-In, and morphologically by scanning electron microscopy. Platelet mv adhered to subendothelium quantitatively over time. Using a modified Baumgartner chamber, we found adhesion of mv to subendothelium significantly increased with increasing shear rates. With this same model we found that prior exposure of subendothelium to mv greatly increased subsequent adhesion of platelets to the same everted vessel, compared to platelet adhesion in the absence of mv. All of these experiments were conducted with mv suspended in ACD/saline, indicating that plasma components are not essential for adhesion of mv. Our studies show that platelet mv adhere to subendothelium in much the same way as do platelets, and support the concept of a hemostatic role for mv in that they appear to increase platelet adhesion.

Animals↗
PubMed1992

[Bilateral epidural hematomas].

Bilateral epidural haematomata are rare and comprise only approximately 2% of all the intracranial haematomata. The symptomatology frequently differs from that of unilateral haematomata in that lateralisation is frequently absent. Most commonly, protracted or secondary development of clouding of consciousness is encountered. In cases of head injury, the above mentioned symptoms are indications for emergency computed tomographic (CT) scanning. When operation is performed early, the prognosis is good.

Adult↗
PubMed1992

Paired comparison of platelet concentrates prepared from platelet-rich plasma and buffy coats using a new technique with 111In and 51Cr.

Two techniques for the preparation of platelet concentrate (PC), the standard platelet-rich plasma (PRP) and buffy coat (BC) methods, were compared in nine paired studies with regard to platelet harvest, white cell (WBC) contamination, and PC quality after 5 days of 22 degrees C storage. Platelet harvest using the BC method averaged approximately 56 percent of the whole blood level (6.2 x 10(10)/concentrate), which was less than the 76 percent achieved with the PRP-PC method (8.7 x 10(10)/concentrate). An additional 5 units collected into an experimental siphon bag for BC-PC processing showed improved platelet harvest (6.7 x 10(10)/concentrate, or approx. 70% of whole blood). WBCs remaining in the BC-PC averaged 0.19 x 10(8) per unit compared to 3.6 x 10(8) per unit for PRP-PC. Buffy coat processing produced red cell (RBC) units with 50 percent of the WBC contamination of conventionally prepared units (9.8 +/- 6.2 x 10(8)/unit vs. 18.9 +/- 7.1 x 10(8)/unit). The siphon bag further reduced WBC levels in the AS-3 RBC units (6.4 +/- 3.7 x 10(8)/unit). In vitro studies performed on Days 1 and 5 after collection showed no significant differences in platelet metabolic and biologic function or cell integrity. Beta-thromboglobulin and surface glycoprotein levels, indicators of platelet activation and membrane alteration, respectively, did not differ significantly in the PRP-PC and BC-PC; nor was lactate production higher in PRP-PC, despite the substantially higher WBC counts. Autologous in vivo platelet viability determinations were performed by using concurrent transfusion of 111In-labeled freshly drawn platelets and 51Cr-labeled stored platelets.(ABSTRACT TRUNCATED AT 250 WORDS)

Blood Platelets↗
PubMed1992

In vitro and in vivo evaluation of cotton wool filtration of platelet concentrates obtained by automated and manual apheresis.

The effect of cotton wool filtration of apheresis platelet concentrates (PCs) on platelet viability and complement activation was evaluated by two laboratories. PCs were prepared by automated (Lab A, n = 5) or manual (Lab B, n = 5) apheresis. After storage for 1 day, the PC was filtered through cotton wool before transfusion on one occasion and, on the other occasion, filtered through a standard screen filter before transfusion to the same donor. Five paired studies were performed by each laboratory. Except for a small, but significant reduction in mean platelet size, from 7.3 +/- 1.1 to 6.6 +/- 0.9 microns 3, after cotton wool filtration, no effect of filtration on various tests of in vitro platelet function and morphologic integrity was found. As demonstrated by autologous radiolabeled studies, no effect of cotton wool filtration on platelet viability was found by Laboratory B, while Laboratory A found a slight increase in the percentage of recovery from 59 +/- 4 to 68 +/- 13 percent, and a small reduction in survival, from 8.2 +/- 0.9 to 7.7 +/- 0.5 days after cotton wool filtration (p less than 0.05). Cotton wool filtration was associated with a slight increase in C3a levels found in manual apheresis PCs. Neither laboratory found any effect of cotton wool filtration per se on the recipients' white cell (WBC) counts or C3a and C5a levels after transfusion.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Post-transfusion recovery of function of 5-day stored platelet concentrates.

Platelets show a rapid reduction in their responsiveness to aggregating agents during storage for transfusion, but little is known about reversal of this defect in vivo after transfusion. In this study, fresh and stored platelets from the same donor (n = 12) were labelled with 111In or 51Cr, respectively, mixed, and simultaneously infused. Blood samples were taken for up to 5 d post-infusion, and the functional behaviour of the labelled platelets ex vivo was measured by retention on glass bead columns, and by whole blood aggregability to ADP, epinephrine and ristocetin. Aggregation was determined by filtering aggregated samples through a column of cotton wool to remove the aggregates, and quantitated as per cent decrease in radioactive counts. The study showed that, although infused radiolabelled 5 d stored platelets had a significantly lower aggregability towards ADP and epinephrine immediately (1 h) after infusion (32% and 29%, respectively, of fresh platelet values), a complete restoration to fresh platelet levels was found 24-72 h post-infusion, with no further change observed over the ensuing 5 d with either fresh or stored labelled platelets. A slightly (6-9%) lower adhesion to both uncoated and collagen-coated beads was found for the stored platelets throughout the 5 d period of study post-infusion. In conclusion, these studies show that, with ex vivo testing, platelets stored for 5 d quickly recovered adhesion and aggregability capabilities similar to that of fresh platelets, suggesting that the functional lesion developed during storage is quickly and completely reversed after infusion.

Adenosine Diphosphate↗