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1.  Comparison of plateletpheresis on three continuous flow cell separators 
Platelet concentrate (PC) remains one of the most important support measures in thrombocytopenic patients. An efficient cell separator is a prerequisite for an optimally functioning apheresis setup. Donor blood count may undergo a temporary reduction after the procedure.
The aim was to find the extent of reduction in donor blood count (hemoglobin, hematocrit, white blood cell, and platelet) after plateletpheresis and to evaluate the cell separator for collection efficiency, processing time, and leukoreduction.
Study Design and Methods:
Two hundred and thirty seven procedures performed on the Amicus (N = 121), Fenwal CS-3000 Plus (N = 50) and Cobe spectra (N = 66) in a one year period were evaluated. The procedures performed on the continuous flow centrifugation (CFC) cell separators and donor blood counts (pre and post donation) done were included in the study.
The percent reduction in hemoglobin (HB), hematocrit (HCT), white blood cell (WBC) and platelet count ((PLT ct) was 2.9, 3.1, 9, 30.7 (Mean, N = 237) respectively after the procedure. The post donation PLT ct reduced to < 100×109/L (range 80-100) in five donors (N = 5/237, Amicus). The pre donation PLT ct in them was 150-200×109/L. Collection efficiency (percent) of Amicus (79.3) was better as compared to the other two machines (CS: 62.5, Cobe: 57.5). PC collected on Cobe spectra had <1×106 WBC. The donor pre donation PLT levels had a positive correlation to the product PLT yield (r = 0.30, P = 0.000).
Monitoring donor blood counts helps to avoid pheresis induced adverse events. A cautious approach is necessary in donors whose pre donation PLT ct is 150-200×109/L. The main variable in PLT yield is donor PLT ct (pre donation). High collection efficiency is a direct measure of an optimally functioning cell separator.
PMCID: PMC2920476  PMID: 20808650
Continuous flow cell separator; donor blood count; plateletpheresis; platelet yield; Blood donor; apheresis
2.  Platelet Recruitment during Multiple Donor Platelet Apheresis Differs between Cell Separators 
Recruitment of platelets (PLT) during donor PLT apheresis may facilitate the harvest of multiple units within a single donation.
We compared two PLT apheresis procedures (Amicus and Trima Accel) in a prospective, randomized, paired cross-over study in 60 donors. The 120 donations were compared for depletion of circulating PLT in the donors, PLT yields and PLT recruitment. A recruitment was defined as ratio of total PLT yield and donor PLT depletion > 1.
Despite comparable differences of pre- and post-apheresis PLT counts (87 × 109/l in Trima Accel vs. 92 × 109/l in Amicus, p = 0.383), PLT yields were higher with Trima Accel (7.48 × 1011 vs. 6.06 × 1011, p < 0.001), corresponding to a higher PLT recruitment (1.90 vs. 1.42, p < 0.001). We observed a different increase of WBC counts after aphereses, which was more pronounced with Trima Accel than with Amicus (1.30 × 109/l vs. 0.46 × 109/l, p < 0.001).
Both procedures induced PLT recruitment. This was higher in Trima Accel, contributing to a higher yield in spite of a comparable depletion of circulating PLT in the donors. This recruitment facilitates the harvest of multiple units within a single donation and seems to be influenced by the procedure utilized. The different increases of circulating donor white blood cells after donation need further investigation.
PMCID: PMC3128152  PMID: 21760764
Platelet apheresis; Platelet recruitment; Cell separator
3.  Pre- and post- donation haematological values in healthy donors undergoing plateletpheresis with five different systems 
Blood Transfusion  2009;7(3):188-192.
Although automated cell separators have undergone a lot of technical refinements, attention has been focused more on the quality of platelet concentrates than on donor safety. We planned this prospective study to observe the effects of automated plateletpheresis on normal haematological values of healthy donors and to determine whether the haematological alterations had any clinical consequences.
Study design and methods
The study was conducted on 457 healthy, first-time plateletpheresis donors over a period of 26 months. The plateletpheresis procedures were performed using five different cell separators and various pre- and post-donation haematological values such as haemoglobin concentration (Hb), haematocrit (Hct), platelet and white blood cell (WBC) counts, mean platelet volume and platelet distribution width were measured in all donors.
We observed that the Hb, Hct, platelet and WBC counts decreased significantly in the donors (p<0.01) after each procedure, without there being significant changes in mean platelet volume or platelet distribution width. The decreases in Hb and Hct were significantly greater with the CS 3000 and Amicus machines, while the decreases in platelet and WBC counts were significantly greater with the CS 3000 and Fresenius separators.
Although a significant drop in complete blood count was observed in all donors, none manifested features of thrombocytopenia or anaemia. Nevertheless, more prospective studies on this aspect are required in order to establish guidelines for donor safety in apheresis and also to help in assessing donor suitability, especially given the present trend of double product apheresis collections.
PMCID: PMC2719270  PMID: 19657482
Plateletpheresis; haematological values; cell separator; donor safety; platelet count
4.  Initial Values of Donor Hematocrit and Efficiency of Plateletpheresis 
Acta Informatica Medica  2013;21(2):116-119.
The collection of platelets by apheresis is considered as a very great progress in transfusion medicine. A larger yield (total number of collected platelets) is obtained if the donor has a greater number of initial platelets and if the separation is done in a shorter time. One of the parameters is also the efficiency of the platelet collection (expressed in percentage) on the value of which different factors may have direct or indirect influence.
To calculate the efficiency of platelet collection with the separator Fenval Baxter AMICUS and to compare the efficiency of platelet collection with this separator in relation to the initial value of donor hematocrit.
Donors and Methods
The donors who participated in this study were divided into groups according to the value of the donor’s ‘hematocrit before separation. Group C consisted of donors whose initial value of the hematocrit was lower or equal to 46%. Group D consisted of donors whose initial value of the hematocrit was higher than 46%. The process was carried out on Fenval Baxter AMICUS. The expected efficiency of the collection was obtained by dividing the total number of collected cells by the expected total number of processed cells, i.e. the total number of cells passed through the equipment.
In the 258 separations which satisfied the fixed criteria were men in 226 cases (87.6%) and women in 32 (12.4%). There is a statically significant difference in the platelet value between the groups and this value is higher in group D than in group C. The average value of platelets before separation was 46.66. The range of minimal and maximal value is from 38.8 to 52.4 ±2.78. The initial value of hematocrits of the donor does not intervene in the length of the separation, but it has a significant effect on the efficiency of the platelet collection. Increases in the number of hematocrits significantly decrease the efficiency of platelet collection. In practice it means that we can base on this fact make a better selection of donors. In this kind selection, one should prefer a donor with a higher number of initial platelets and lower levels of hematocrits. In that way we can collect a more important yield, have a shorter length of separation and increase the efficiency of platelet collection. Its advantage is as well medical because of a more important yield but also financial because of the decrease of the length of the separation and the increase in efficiency. Key words: value of hematocrits, donors, apheresis, platelets, efficiency.
PMCID: PMC3766539  PMID: 24058252
value of hematocrits; donors; apheresis; platelets; efficiency
5.  Evaluation of White Blood Cell- and Platelet-Derived Cytokine Accumulation in MIRASOL-PRT-Treated Platelets 
Soluble mediators in platelet concentrates (PCs) released from contaminating white blood cells (WBCs) and platelets (PLTs) themselves are supposed to promote allergic and non-hemolytic febrile transfusion reactions in the recipient. Pathogen reduction technologies (PRTs) prevent replication and proliferation of pathogens as well as of WBCs, and may reduce cytokine accumulation in PCs during storage and prevent adverse events after PLT transfusion. On the other hand, such treatments may also lead to increased cytokine production by stimulation of WBCs or PLTs due to the photochemical or photodynamical process itself.
Material and Methods
12 triple-dose PLT apheresis collections were leukoreduced by the process-controlled leukoreduction system of the Trima Accel machine and split into 3 units undergoing Mirasol-PRT treatment (M) or gamma irradiation (X) or remaining untreated (C). During storage for up to 7 days, PLT activation, WBC-derived Th-1/2, and inflammatory as well as PLT-derived cytokines were measured by cytometric bead array and enzymelinked immunosorbent assay, respectively.
Independent of treatment, all PLT products exhibited low levels of WBC-associated cytokines near or below assay detection limits. WBC-associated cytokines were not elevated by Mirasol-PRT treatment. PLT-derived cytokines were detected at higher levels and increased significantly during storage in all units. Most likely due to higher PLT activation, M units showed significantly higher levels of PLT-derived cytokines compared to untreated and gamma-irradiated units on day 5 of storage.
In all PCs, PLTs themselves were the main source of cytokine release. Mirasol-PRT treatment was associated with a significantly increased PLT activation and accumulation of PLT-derived cytokines during storage, without affecting WBC-derived cytokines relative to controls.
PMCID: PMC2928824  PMID: 20823992
Pathogen reduction; Mirasol-PRT; Ccytokines; Transfusion reaction
6.  Red Cell Apheresis with Automated In-Line Filtration 
The aim of this study was to provide data on concurrent red blood cell (RBC) and platelet (PLT) apheresis with RBC in-line leukoreduction and automated addition of saline-adenine-glucose-mannitol (SAGM) using the new version (V6.0) of Trima Accel®.
In this two-center paired study, each subject completed a test and a control procedure with an interval of 9 weeks between procedures. In the test arm, single RBC and PLT units were collected on the Trima Accel V6.0 (in-line leukofiltration and automated addition of SAGM). In the control arm, they were collected on Trima Accel V5.1/V5.2 (post-collection leukoreduction, manual SAGM addition). RBC percent hemolysis, potassium concentration and adenosine triphosphate over storage, hemoglobin (Hb) yield, and residual white blood cells (WBC) were determined.
34 subjects successfully completed both test and control procedures. Post-storage hemolysis was similar in both groups, and all values were less than 0.8% for both arms. Residual WBC counts in all RBC units were less than 1 × 106/unit. In-line processed RBC units (V6.0) have a significantly higher volume and more Hb/unit due to filtration recovery improvements. All procedures were well tolerated by the subjects.
In-line filtration and automated addition of storage solution on Trima Accel V6.0 allows collection of ready-to-use RBC units that meet EU requirements.
PMCID: PMC4025052  PMID: 24847185
Red blood cell; Apheresis; In-line filtration; Leukoreduction; Multicomponent collection; Blood storage
7.  Therapeutic efficacy of different types of platelet concentrates in thrombocytopenic patients 
Platelet Rich Plasma-Platelet concentrate (PRP-PC), Buffy Coat poor-platelet concentrate (BCPC), and Apheresis — PC were prepared and their therapeutic efficacy were assessed in thrombocytopenic patients.
Study design and methods
PRP-PC and BC-PC were prepared from whole blood and Apheresis-PC by automated cell separator. The post transfusion efficacy of transfused platelets was assessed at 1 hour and 20 hours by corrected count increment (CCI) and percentage recovery (PR).
A total of 60 patients’ (20 each for PRP-PC, BC-PC and Apheresis-PC) were enrolled in this study. Forty one patients received therapeutic and nineteen received prophylactic transfusion support. Patients with aplastic anemia 43% (25/60) and acute leukemia 38% (23/60) formed a majority of study population. Platelet dosage of patients’ received PRP-PC, BC-PC and apheresis-PC were 2.4±0.82 × 1011 (mean±SD), 2.2±0.83 × 1011 (mean±SD) and 4.14±1.82 × 1011 (mean±SD) and ranged from 1.16–4.11 × 1011, 1.04−4.20 × 1011 and 1.22−8.90 × 1011 respectively. There was significantly increase in inter-transfusion interval with Apheresis-PC than with PRP-PC and BC-PC recipients [(Mean±S.D.), 4.7±1.33 days Vs 2.7±0.82 days Vs 2.5±0.7 days respectively] (p < 0.05).
Patients transfused with apheresis-PC had received higher platelet dosage than PRP-PC and BC-PC and this difference was statistically significant (p < 0.001). The post transfusion platelet counts and increments at 1 hour and 20 hours were significantly higher with apheresis-PC than PRP-PC and BC-PC (p < 0.001). However, the corrected count increment (CCI) and percentage recovery (PR) in all three groups were comparable. There was significantly increase in inter-transfusion interval with apheresis-PC than PRPPC and BC-PC (p < 0.05).
PMCID: PMC3453166  PMID: 23100935
Random donor platelets; Buffy coat poor-platelet concentrate; Platelet Rich Plasma-Platelet concentrate; Thrombocytopenic patients
8.  Platelet count is a sensitive predictor of autologous peripheral blood progenitor cell collection yield in previously treated plasma cell disease patients 
Transfusion  2008;48(6):1106-1114.
It is often a clinical dilemma to determine when to collect autologous peripheral blood progenitor cells (PBPCs) in patients who received prior chemotherapy. It is also challenging to predict if the collected cells will be enough for one or two transplants.
A total of 103 PBPC donors were followed to evaluate factors that predict poor autologous PBPC collection. The donors were categorized into three groups: plasma cell disorders (PCDs), lymphomas, and normal allogeneic donors.
Our evaluation showed that platelet (PLT) count before growth factor administration significantly correlated with total CD34+ cell yield (Spearman r = 0.38, p < 0.001). Further analysis showed this correlation was only significant in plasma cell disease patients who received prior chemotherapy (Spearman r = 0.5, p = 0.008). Baseline PLT counts did not correlate with PBPC collection yield in untreated PCD, lymphoma, and normal allogeneic donors. In addition, daily PLT count during PBPC harvest correlated with CD34+ cell yield for that day (Spearman r = 0.41, p < 0.001). With a multiple linear regression model (adjusted R2 = 0.31, AIC = 63.1), it has been determined that the baseline PLT count significantly correlates with total CD34+ cell yield in treated PCD patients.
Baseline PLT count is a sensitive indicator of autologous PBPC mobilization in PCD patients who received prior chemotherapy. This finding may be considered before growth factor administration to determine the optimal period to mobilize treated PCD patients and to predict if enough cells can be collected for one or two transplants.
PMCID: PMC3919131  PMID: 18315528
9.  Probiotic-associated high-titer anti-B in a group A platelet donor as a cause of severe hemolytic transfusion reactions 
Transfusion  2009;49(9):1845-1849.
Hemolytic transfusion reactions (HTRs) can occur with transfusion of platelets (PLTs) containing ABO-incompatible plasma. Reported cases have involved group O donors. Two cases of PLT-mediated HTRs associated with the same group A plateletpheresis component, collected from a donor taking high doses of probiotics are reported.
Case 1 was a 40-year-old 69-kg group B stem cell transplant patient who received one-half of a group A plateletpheresis component. Severe back pain occurred 10 minutes into the transfusion, accompanied by anemia and hyperbilirubinemia. Case 2 was a 5-year-old 26-kg group B male with aplastic anemia who received the other half of the same plateletpheresis component, volume reduced to 37 mL. Syncope occurred immediately after the transfusion, with laboratory evidence of hemolysis a few hours later.
Serologic investigation of posttransfusion samples from both patients revealed positive direct anti-globulin tests: C3d only for Case 1 and immunoglobulin (Ig)G and C3d for Case 2; the eluates contained anti-B. The group A donor’s anti-B titer was 16,384 at saline and IgG phases. Donor lookback revealed that the donor had donated 134 apheresis PLTs over many years. For 3 years, he had intermittently taken probiotics; 3 weeks before the index donation, he began taking three tablets of probiotics every day. Lookback of prior group B recipients uncovered a case of acute hemolysis that was not recognized at the time. The solubilized probiotic inhibited anti-B in vitro.
Non–group O PLT donors can have high-titer anti-A or anti-B that might mediate HTRs, and probiotic ingestion in blood donors represents a novel mechanism of stimulating high-titer anti-B.
PMCID: PMC3421026  PMID: 19453987
10.  Thrombocytosis is a significant indictor of hypercoagulability, prognosis and recurrence in gastric cancer 
Although thrombocytosis has been reported in a variety of cancer types, the standard of thrombocytosis in gastric cancer (GC) and the association between thrombocytosis and the clinicopathological features of patients with GC remain unclear. In the present study, 1,763 GC patients were retrospectively filtered by preoperative thrombocytosis and compared with control group A (n=107) that had benign gastric lesions and control group B (n=100) that were GC patients with a normal platelet (PLT) count. Associations between clinical variables and preoperative PLT counts were assessed by univariate and multivariate analyses. Kaplan-Meier survival curves and Cox regression were used to evaluate the effect of thrombocytosis on prognosis. Sensitivities and specificities of the PLT counts in predicting recurrence were analyzed via area under the receiver operating characteristic curve (AUROC). The results indicated that the incidence of thrombocytosis in GC patients was higher than in benign gastric lesion patients, with 4.03% of GC patients having a PLT count >400×109/l (P=0.014) and 12.08% had a PLT count >300×109/l (P<0.001). For the patients with a PLT count >400×109/l, the frequency of abnormal PLT counts in GC correlated with tumor size (P<0.001), tumor, node and metastasis (TNM) classification (P=0.002), invasive degree (P=0.003) and D-dimer (P=0.013) and fibrinogen concentrations (P=0.042). Tumor size (P=0.002), TNM classification (P<0.001) and depth of penetration (P=0.001) were independent factors for thrombocytosis. However, thrombocytosis functioned as an independent prognostic factor for GC patients with a PLT count >400×109/l (relative risk, 1.538; 95% confidence interval, 1.041–2.271). In the majority of patients (17/24) with a high preoperative PLT count that decreased to a normal level following resection, PLT levels increased again at recurrence. Sensitivities and specificities of thrombocytosis for recurrence in those patients were 70.8 and 83.3%, respectively (AUROC, 0.847; P=0.01). Therefore, a PLT count of 400×109/l is a suitable threshold for defining thrombocytosis in GC. Thrombocytosis was shown to affect the blood hypercoagulable state and also have a negative prognostic value for GC patients. PLT monitoring following surgery was useful to predict the recurrence for specific GC patients that suffered preoperative thrombocytosis but had restored PLT levels following resection.
PMCID: PMC4061185  PMID: 24944610
gastric cancer; thrombocytosis; recurrence; survival analysis
11.  Initial performance evaluation of the UniCel® DxH 800 Coulter® cellular analysis system 
The Beckman Coulter UniCel® DxH 800 is a hematology analyzer incorporating new electronic and mechanical design with advanced algorithm technology to perform CBC, white blood cell (WBC) differential, nucleated red blood cell (NRBC), and reticulocyte analysis. Evaluation of this instrument was performed in our 800-bed tertiary care hospital and specifically centered upon the correlation of WBC, NRBC, and platelet (PLT) enumeration when compared to a predicate analyzer, the Coulter® LH 780, and flow cytometry (FCM) reference methods. Of particular interest were those samples with morphologically confirmed interference and extreme leukocytosis (evaluated with respect to red blood cell parameter correction). The sample set (n = 272) consisted of morphologically normal and hematologically abnormal patients. Correlation of the WBC, PLT, and NRBC showed r2 values of 0.994, 0.985, and 0.910 for the DxH 800 vs. FCM, respectively. The presence of interfering particles did not affect the accuracy of the DxH 800 with respect to WBC counts. The DxH 800 showed accurate PLT and NRBC counts in the clinically significant low range when compared to FCM. Compared to the LH 780, flagging rates were significantly reduced (NRBC flag), or equivalent (WBC, PLT flag) on the DxH 800. The DxH 800 demonstrated higher sensitivity and specificity for PLTs and NRBCs and achieved a lower NRBC false negative rate compared to the LH 780. The UniCel® DxH 800 represents a significant improvement to previous impedance analyzers in accurately detecting the presence of NRBCs at counts >1/100 WBC. Furthermore, it provides accurate PLT and WBC counts in the presence of interference and improved NRBC flagging efficiency when compared to the LH 780. Correction of red blood cell parameters is appropriate and accurate in cases of extreme leukocytosis.
PMCID: PMC3044820  PMID: 20491996
Hematology analyzer; WBC; NRBC; platelet count; extreme leukocytosis; interference
12.  Blood donations from previously transfused or pregnant donors: a multicenter study to determine the frequency of alloexposure 
Transfusion  2010;51(6):1197-1206.
Transfusion-related acute lung injury (TRALI) mitigation strategies include the deferral of female donors from apheresis platelet (PLT) donations and the distribution of plasma for transfusion from male donors only. We studied the implications of these policies in terms of component loss at six blood centers in the United States.
We collected data from allogeneic blood donors making whole blood and blood component donations during calendar years 2006 through 2008. We analyzed the distribution of donations in terms of the sex, transfusion and pregnancy histories, and blood type.
A TRALI mitigation policy that would not allow plasma from female whole blood donors to be prepared into transfusable plasma components would result in nearly a 50% reduction in the units of whole blood available for plasma manufacturing and would decrease the number of type AB plasma units that could be made from whole blood donations by the same amount. Deferral of all female apheresis PLT donors, all female apheresis PLT donors with histories of prior pregnancies, or all female apheresis PLT donors with histories of prior pregnancies and positive screening test results for antibodies to human leukocyte antigens (HLAs) will result in a loss of 37.1, 22.5, and 5.4% of all apheresis PLT donations, respectively.
A TRALI mitigation policy that only defers female apheresis PLT donors with previous pregnancies and HLAs would result in an approximately 5% decrease in the inventory of apheresis PLTs, but would eliminate a large proportion of components that are associated with TRALI.
PMCID: PMC3606016  PMID: 21182532
13.  The association between red blood cell and platelet transfusion and subsequently developing idiopathic pneumonia syndrome after hematopoietic stem cell transplantation 
Transfusion  2013;54(4):1071-1080.
Blood transfusions are common during hematopoietic stem cell transplantation (HSCT) and may contribute to lung injury.
This study examined the associations between red blood cell (RBC) and platelet (PLT) transfusions and idiopathic pneumonia syndrome (IPS) among 914 individuals who underwent myeloablative allogeneic HSCT between 1997 and 2001. Patients received allogeneic blood transfusions at their physicians' discretion. RBCs, PLTs, and a composite of “other” transfusions were quantified as the sum of units received each 7-day period from 6 days before transplant until IPS onset, death, or Posttransplant Day 120. RBC and PLT transfusions were modeled as separate time-varying exposures in proportional hazards models adjusted for IPS risk factors (age, baseline disease, irradiation dose) and other transfusions. Timing of PLT transfusion relative to myeloid engraftment and PLT ABO blood group (match vs. mismatch) were included as potential interaction terms.
Patients received a median of 9 PLT and 10 RBC units. There were 77 IPS cases (8.4%). Each additional PLT unit transfused in the prior week was associated with 16% higher IPS risk (hazard ratio, 1.16; 95% confidence interval, 1.09–1.23; p < 0.001). Recent RBC and PLT transfusions were each significantly associated with greater risk of IPS when examined without the other; only PLT transfusions retained significance when both exposures were included in the model. The PLT association was not modified by engraftment or ABO mismatch.
PLT transfusions are associated with greater risk of IPS after myeloablative HSCT. RBCs may also contribute; however, these findings need confirmation.
PMCID: PMC4059041  PMID: 24033082
14.  Prominent features of platelet count, plateletcrit, mean platelet volume and platelet distribution width in pulmonary tuberculosis 
We aimed to investigate the relation of platelet count (PLT) and plateletcrit (PCT), mean platelet volume (MPV) and platelet distribution width (PDW) with other acute phase reactants and radiological extent in pulmonary tuberculosis (PTB).
One hundred patients with PTB (Group 1), 50 patients with community-acquired pneumonia (Group 2) and 28 healthy control individuals (Group 3) were included in this analytic study.
WBC (White Blood Cell), ESR (Eritrocyte Sedimentation Rate), CRP (C-Reactive Protein), PLT and PCT values were both in Group 1 and Group 2 than in Group 3. PDW values were significantly higher in Group 1 than Group 3. WBC, ESR and CRP values were lower, while PLT and PCT values were higher in the Group 1 compared to Group 2 (p < 0.001). PLT was positively correlated with CRP and ESR values in the tuberculosis group (p < 0.001), while it was not correlated with CRP and ESR in the pneumonia group (p > 0.05). ESR, CRP, PLT and PCT values were found higher in radiological advanced stage (Stage 3) patients with PTB, while hemoglobin (Hb) was found lower (p < 0.05). Higher WBC, ESR, CRP and PCT values as well as radiological advanced stage were more common in PTB patients with thrombocytosis compared to the patients with normal platelet count, whereas Hb was found lower in these patients.
This study indicates that reactive thrombocytosis and higher PCT and PDW develop frequently in PTB and there is a relation between thrombocytosis and acute phase reactants, that is the inflammatory response. In addition, tuberculosis with radiological advanced stage is seen more frequently in the patients with thrombocytosis and higher PCT, drawing attention to the possible role of platelets in the cell-based immune process of tuberculosis.
PMCID: PMC3529701  PMID: 23114411
Mean platelet volume; Platelet; Plateletcrit; Platelet distribution width; Pneumonia; Pulmonary tuberculosis; Thrombocytosis
15.  Effect of storage on levels of nitric oxide metabolites in platelet preparations 
Transfusion  2012;53(3):637-644.
Nitric oxide (NO), a potent signaling molecule, is known to inhibit platelet function in vivo. We investigated how the levels of NO and its metabolites change during routine platelet storage. We also tested whether the material of platelet storage containers affects nitrite content since many plastic materials are known to contain and release nitrite.
Study design and methods
For nitrite and nitrate measurement, leukoreduced apheresis platelets (PLT) and concurrent plasma (CP) were collected from healthy donors using the Trima Accel. Sixty mL aliquots of PLT or CP were stored in CLX or PL120 Teflon containers at 20–24°C with agitation and daily samples were processed to yield PLT pellet and supernatant. In a separate experiment, PLT was stored in PL120 Teflon to measure NO generation using electron paramagnetic resonance (EPR).
Nitrite level increased markedly in both PLT supernatant and CP stored in CLX containers at a rate of 58 nM/day and 31 nM/day respectively. However, there was a decrease in nitrite level in PLT stored in PL120 Teflon containers. Nitrite was found to leach from CLX containers and this appears to compensate for nitrite consumption in these preparations. Nitrate level did not significantly change during storage.
Platelets stored at 20–24°C maintain measurable levels of nitrite and nitrate. Nitrite decline in non-leachable Teflon containers in contrast to increases in CLX containers which leach nitrite, suggests that it is consumed by platelets, residual leukocytes or erythrocytes. These results suggest NO-related metabolic changes occur in platelet units during storage.
PMCID: PMC3548060  PMID: 22804724
nitric oxide; nitrite; platelet storage; transfusion
16.  The determinants of granulocyte yield in 1198 granulocyte concentrates collected from unrelated volunteer donors mobilized with dexamethasone and granulocyte–colony-stimulating factor: a 13-year experience 
Transfusion  2008;49(3):421-426.
The combination of granulocyte–colony-stimulating factor (G-CSF [filgrastim]) and dexamethasone (G-CSF/dex) is an effective granulocyte mobilization regimen, but the variables that affect donor neutrophil response and granulocyte collection yield are not well characterized.
A computerized database containing records of 1198 granulocyte collections from 137 unrelated volunteer apheresis donors during a 13-year period was retrospectively analyzed. Donors were categorized by age, sex, and cumulative number of granulocyte donations. Complete blood counts at baseline and after G-CSF/dex stimulation were recorded. The outcome variables include the pre-procedure absolute neutrophil count (preANC), which reflects G-CSF/dex stimulation, and the granulocyte product yield per liter processed (BagGranYield/L).
Higher baseline ANC and platelet (PLT) counts were significantly associated with higher preANC while a larger number of prior granulocytapheresis procedures was associated with lower preANC. Total filgrastim dose (used in weight-based dosing) did not significantly impact preANC or the granulocyte yield; weight-based dosing at 5 μg per kg and a uniform 480-μg dose produced equivalent preANC. PreANC and weight were the key determinants of granulocyte yield (BagGranYield/L).
Apheresis donors with higher baseline PLT counts and ANCs have higher ANCs after G-CSF/dex stimulation; donor age, weight, and sex do not have a significant impact. A uniform G-CSF dose of 480 μg is as effective as weight-based dosing at 5 μg per kg. Donor ANC monitoring should be considered after serial granulocytapheresis procedures.
PMCID: PMC3421027  PMID: 19040597
17.  Efficacy and Side Effects of Granulocyte Collection in Healthy Donors 
We report on the efficacy and side effects of granulocyte collection, which is comparatively infrequently performed in Germany.
Data from 378 healthy donors who underwent 914 granulocyte collections between 1999 and 2007 were retrospectively analyzed. Donors received G-CSF (lenograstim) at a median dose of 5.58 (3.25–7.36) μg/kg body weight with (n = 243) or without (n = 57) 4 mg dexamethasone. Side effects were recorded by donor monitoring and interview (questionnaire).
The median granulocyte yield in apheresis products was 8.47 × 1010 (3.07–14.92 × 1010). Granulocyte yields correlated significantly with gender, baseline WBC, PMN and PLT counts, and nicotine consumption. Dexamethasone and lenograstim administration was more effective than lenograstim administration alone (p < 0.001). Side effects of granulocyte mobilization were generally mild: bone pain in 31.4%, headache in 19.6%, and fatigue in 15.7% of donors. During follow-up (4 weeks), pruritus and/or exanthema were reported in 17.6% of donors.
Granulocyte mobilization with lenograstim with or without dexamethasone was a safe and effective regimen for granulocyte mobilization. Side effects were tolerable and milder than those seen in peripheral blood stem cell donors. Long-term monitoring of granulocyte donors is important to establish optimal standards for the procedure.
PMCID: PMC3776445  PMID: 24179474
Granulocyte collection; Allogeneic donors; G-CSF
18.  Annexin V Release and Transmembrane Mitochondrial Potential during Storage of Apheresis-Derived Platelets Treated for Pathogen Reduction 
In vitro function of stored platelet (PLT) con-centrates was analyzed after applying two different techniques of pathogen reduction technology (PRT) treatment, which could increase cellular injury during processing and storage.
Nine triple-dose PLT apheresis donations were split into 27 single units designated to riboflavin-UVB (M) or psoralen-UVA (I) treatment or remained untreated (C). Throughout 8 days of storage, samples were analyzed for annexin V release, the mitochondrial transmembrane potential (Δψ) and some classical markers of PLT quality (pH, LDH release, hypotonic shock response (HSR)).
PLT count and LDH release of all units maintained initial ranges. All units exhibited a decrease in pH and HSR and an increase in annexin V release and Δψ disruption. Notably, throughout the entire storage period, annexin V release re-mained lowest in M units. Throughout 7 days of storage, M units remained comparable to C units (p > 0.05), whereas inferior values were observed with I units. Here, differences to C units reached significance by day 1 (pH: p < 0.0001), day 5 (annexin V release: p < 0.014), and day 7 (HSR, Δψ: p ≤ 0.003). After PRT treatment, annexin V release and Δψ disruption were significantly (p < 0.001) correlated with pH and HSR.
During storage, all units showed a de-crease in HSR and an increase in acidity, annexin V release and Δψ disruption. While M units remained comparable to C units, I units demonstrated significantly inferior values during terminal storage. This could have resulted from differences in PRT treatment or simply be due to differences in storage media and should be analyzed for clinical relevance in future investigations.
PMCID: PMC2914403  PMID: 20737011
Pathogen reduction technology; Platelet in vitro function; Endogenous annexin V; Transmembrane mitochondrial potential; INTERCEPT BLOOD SYSTEM; MIRASOL-PRT
19.  Adverse events associated with apheresis procedures: Incidence and relative frequency 
Apheresis procedures [Plateletpheresis, Plasmapheresis/ Therapeutic Plasma Exchange (TPE), & Peripheral Blood Stem Cell Collection (PBSC)] are usually well tolerated. Occasionally, Adverse Events (AEs) of variable severity may occur during or after the procedure. AEs that occur in Donors/Patients are divided into local reactions and systemic reactions.
Materials and Methods:
A total of 3,367 apheresis procedures were performed, out of which 3,120 were plateletpheresis procedures, and out of which 1,401 were on Baxter CS 3000 & 1,719 were on Haemonetics MCS+ cell separators. Rest of 247 TPE & PBSC procedures were done on Haemonetics MCS+ cell separators.
90 AEs were reported in relation to the 3,367 procedures. Out of 90 AEs, 85 AEs (94%) were associated with plateletpheresis (n = 3,120) and 05 AEs (06%) with TPE & PBSC (n = 247). The rate of vascular injury (VI), Citrate reaction (CR), and Presyncopal/Syncopal (PS/S) in plateletpheresis was 1.6% (52/3,120), 0.96% (30/3,120), and 0.096% (03/3,120), respectively. The rate of CR in TPE and PBSC was 1.23% (02/162) and 2.3% (02/85), respectively. The rate of PS/S in PBSC was 1.17% (01/85). AEs for Plateletpheresis, TPE & PBSC were 2.7% (85/3,120), 1.23% (02/162), and 3.5% (03/85), respectively. VI, CR, and PS/S were mostly of mild intensity. Both cell separators were equally safe, when AEs associated with plateletpheresis were compared with each other; 2.8% on CS 3000 & 2.6% on MCS+.
Apheresis procedures performed on cell separators are safe, with a low incidence of significant AEs. No significant difference was noted in AEs among the two cell separators studied.
PMCID: PMC3613659  PMID: 23559763
Adverse events; citrate reaction; peripheral blood stem cell; presyncopal/syncopal; therapeutic plasma exchange; vascular injury
20.  Comparison of total splenic artery embolization and partial splenic embolization for hypersplenism 
AIM: To evaluate whether total splenic artery embolization (TSAE) for patients with hypersplenism delivers better long-term outcomes than partial splenic embolization (PSE).
METHODS: Sixty-one patients with hypersplenism eligible for TSAE (n = 27, group A) or PSE (n = 34, group B) were enrolled into the trial, which included clinical and computed tomography follow-up. Data on technical success, length of hospital stay, white blood cell (WBC) and platelet (PLT) counts, splenic volume and complications were collected at 2 wk, 6 mo, and 1, 2, 3, 4 years postoperatively.
RESULTS: Both TSAE and PSE were technically successful in all patients. Complications were significantly fewer (P = 0.001), and hospital stay significantly shorter (P = 0.007), in group A than in group B. Post-procedure WBC and PLT counts in group A were significantly higher than those in group B from 6 mo to 4 years (P = 0.001), and post-procedure residual splenic volume in group A was significantly less than that observed in group B at 1, 2, 3 and 4 years post-procedure (P = 0.001). No significant differences were observed in red blood cell counts and liver function parameters between the two groups following the procedure.
CONCLUSION: Our results indicate that TSAE for patients with hypersplenism not only delivers a better long-term outcome, but is also associated with lower complication rates and a shorter hospital stay than PSE.
PMCID: PMC3386328  PMID: 22791950
Embolization; Hypersplenism; Complications; White cell counts; Platelet counts
21.  Long-term dynamics of hematological data and spleen volume in cirrhotic patients after liver transplantation-various dynamics depending on etiology 
SpringerPlus  2013;2:374.
Hypersplenism is a common complication in cirrhotic patients, and liver transplantation would be one of the effective treatments. However, detailed dynamics, especially over a long term, are not fully understood. We investigated the long-term dynamics of hematological data and spleen volumes, as well as their correlation in cirrhotic patients who underwent liver transplantation.
Patients and methods
We studied 53 cirrhotic patients who underwent liver transplantation at our institute and followed for more than 1 year. Hematological data were collected from medical records, while spleen volumes were determined by CT volumetry at 0, 1, 3, 6, 12, 24, 36, 48, 60 postoperative months (POM).
(1) Platelet (Plt) and hemoglobin (Hb) levels were gradually increased up to 18 and 10 POM, respectively, in contrast with white blood cells (WBC), which remained mostly unchanged from pretransplantation levels. (2) Spleen volume was sharply decreased in the first POM, then showed a slower but steady decline up to 48 POM. (3) Spleen volume was significantly correlated with hematological data, though the levels were generally weak (Plt: r = 0.433, p < 0.001; Hb: r = 0.233, p < 0.001; WBC: r = 0.217, p = 0.001). (4) Spleen volume was strongly correlated with all hematological parameters in HBV patients (Plt: r = 0.617, p < 0.0001; Hb: r = 0.401, p < 0.001; WBC: r = 0.387, p < 0.001), in contrast with that in other etiologies, which had generally weak correlations though some were statistically significant.
We investigated the long-term dynamics of hematological data and spleen volume in cirrhotic patients after liver transplantation. Unique dynamics and correlations between them were found among the different etiologies investigated.
PMCID: PMC3755816  PMID: 24010032
Hematological parameters; Spleen volumes; Liver transplantation; Long-term dynamics
22.  Globulin-platelet model predicts minimal fibrosis and cirrhosis in chronic hepatitis B virus infected patients 
AIM: To establish a simple model consisting of the routine laboratory variables to predict both minimal fibrosis and cirrhosis in chronic hepatitis B virus (HBV)-infected patients.
METHODS: We retrospectively investigated 114 chronic HBV-infected patients who underwent liver biopsy in two different hospitals. Thirteen parameters were analyzed by step-wise regression analysis and correlation analysis. A new fibrosis index [globulin/platelet (GP) model] was developed, including globulin (GLOB) and platelet count (PLT). GP model = GLOB (g/mL) × 100/PLT (× 109/L). We evaluated the receiver operating characteristics analysis used to predict minimal fibrosis and compared six other available models.
RESULTS: Thirteen clinical biochemical and hematological variables [sex, age, PLT, alanine aminotransferase, aspartate aminotransferase (AST), albumin, GLOB, total bilirubin (T.bil), direct bilirubin (D.bil), glutamyltransferase, alkaline phosphatase, HBV DNA and prothrombin time (PT)] were analyzed according to three stages of liver fibrosis (F0-F1, F2-F3 and F4). Bivariate Spearman’s rank correlation analysis showed that six variables, including age, PLT, T.bil, D.bil, GLOB and PT, were correlated with the three fibrosis stages (FS). Correlation coefficients were 0.23, -0.412, 0.208, 0.220, 0.314 and 0.212; and P value was 0.014, < 0.001, 0.026, 0.018, 0.001 and 0.024, respectively. Univariate analysis revealed that only PLT and GLOB were significantly different in the three FS (PLT: F = 11.772, P < 0.001; GLOB: F = 6.612, P = 0.002). Step-wise multiple regression analysis showed that PLT and GLOB were also independently correlated with FS (R2 = 0.237). By Spearman’s rank correlation analysis, GP model was significantly correlated with the three FS (r = 0.466, P < 0.001). The median values in F0-F1, F2-F3 and F4 were 1.461, 1.720 and 2.634. Compared with the six available models (fibrosis index, AST-platelet ratio, FIB-4, fibrosis-cirrhosis index and age-AST model and age-PLT ratio), GP model showed a highest correlation coefficient. The sensitivity and positive predictive value at a cutoff value < 1.68 for predicting minimal fibrosis F0-F1 were 72.4% and 71.2%, respectively. The specificity and negative predictive value at a cutoff value < 2.53 for the prediction of cirrhosis were 84.5% and 96.7%. The area under the curve (AUC) of GP model for predicting minimal fibrosis and cirrhosis was 0.762 [95% confidence interval (CI): 0.676-0.848] and 0.781 (95% CI: 0.638-0.924). Although the differences were not statistically significant between GP model and the other models (P all > 0.05), the AUC of GP model was the largest among the seven models.
CONCLUSION: By establishing a simple model using available laboratory variables, chronic HBV-infected patients with minimal fibrosis and cirrhosis can be diagnosed accurately, and the clinical application of this model may reduce the need for liver biopsy in HBV-infected patients.
PMCID: PMC3374981  PMID: 22719186
Globulin; Platelet; Globulin/platelet model; Liver fibrosis; Noninvasive fibrosis biomarker; Chronic hepatitis B virus
23.  Quality assessment of platelet concentrates prepared by platelet rich plasma-platelet concentrate, buffy coat poor-platelet concentrate (BC-PC) and apheresis-PC methods 
Platelet rich plasma-platelet concentrate (PRP-PC), buffy coat poor-platelet concentrate (BC-PC), and apheresis-PC were prepared and their quality parameters were assessed.
Study Design:
In this study, the following platelet products were prepared: from random donor platelets (i) platelet rich plasma - platelet concentrate (PRP-PC), and (ii) buffy coat poor-platelet concentrate (BC-PC) and (iii) single donor platelets (apheresis-PC) by different methods. Their quality was assessed using the following parameters: swirling, volume of the platelet concentrate, platelet count, WBC count and pH.
A total of 146 platelet concentrates (64 of PRP-PC, 62 of BC-PC and 20 of apheresis-PC) were enrolled in this study. The mean volume of PRP-PC, BC-PC and apheresis-PC was 62.30±22.68 ml, 68.81±22.95 ml and 214.05±9.91 ml and ranged from 22-135 ml, 32-133 ml and 200-251 ml respectively. The mean platelet count of PRP-PC, BC-PC and apheresis-PC was 7.6±2.97 × 1010/unit, 7.3±2.98 × 1010/unit and 4.13±1.32 × 1011/unit and ranged from 3.2 –16.2 × 1010/unit, 0.6-16.4 × 1010/unit and 1.22-8.9 × 1011/unit respectively. The mean WBC count in PRP-PC (n = 10), BC-PC (n = 10) and apheresis-PC (n = 6) units was 4.05±0.48 × 107/unit, 2.08±0.39 × 107/unit and 4.8±0.8 × 106/unit and ranged from 3.4 -4.77 × 107/unit, 1.6-2.7 × 107/unit and 3.2 – 5.2 × 106/unit respectively. A total of 26 units were analyzed for pH changes. Out of these units, 10 each were PRP-PC and BC-PC and 6 units were apheresis-PC. Their mean pH was 6.7±0.26 (mean±SD) and ranged from 6.5 – 7.0 and no difference was observed among all three types of platelet concentrate.
PRP-PC and BC-PC units were comparable in terms of swirling, platelet count per unit and pH. As expected, we found WBC contamination to be less in BC-PC than PRP-PC units. Variation in volume was more in BC-PC than PRP-PC units and this suggests that further standardization is required for preparation of BC-PC. As compared to the above two platelet concentrates, all the units of apheresis-PC fulfilled the desired quality control criteria of volume. Apheresis-PC units showed better swirling and platelet count than PRP-PCs and BC-PCs. All the platelet concentrates units had pH well above the recommended norm.
PMCID: PMC2920479  PMID: 20808653
Corrected count increment; buffy coat poor-platelet concentrate; percentage recovery; platelet concentrate; platelet rich plasma-platelet concentrate; random donor platelet; single donor platelets
24.  Electron microscopic observation in case of platelet activation in a chronic haemodialysis subject 
Hematology Reports  2011;3(2):e15.
During haemodialysis (HD), platelets (PLTs) are activated and release granule contents. As HD treatment occurs three times a week, it has been demonstrated that PLTs are exhausted due to the repetitive character of the treatment. To identify PLT depletion morphologically, PLT evaluation was performed by light microscopy and electron microscopy (EM) in a chronic HD subject and a healthy reference subject. Blood samples were taken before the start of HD treatment for measurement of PLT count, PLT volume and size parameters. Blood smears were screened by light microscopy for qualitative evaluation of PLT granule containing cytoplasm, as indicated by its staining density. Morphological PLT parameters of surface area and size of dense bodies were assessed by EM. Data were compared with results of a group of 20 chronic HD subjects and a group of 20 healthy reference subjects. With respect to the percentage of PLTs with appropriate staining density (>75%), light microscopic evaluation showed that this value (9%) was within the range of a group of chronic HD subjects, but considerably below the reference range (70%). EM evaluation revealed an average PLT surface area and dense bodies area of respectively 42% and 31%, if the healthy reference subject was set on 100%. PLTs from a chronic HD subject are considerably smaller and substantially less granular than PLTs from a healthy reference subject. These findings support the hypothesis of PLT depletion in chronic HD subjects due to frequent PLT activation and/or increased urea concentrations.
PMCID: PMC3238486  PMID: 22184536
electron microscopy; platelet activation; platelet degranulation; haemodialysis.
25.  A General Change of the Platelet Transfusion Policy from Apheresis Platelet Concentrates to Pooled Platelet Concentrates is Associated with a Sharp Increase in Donor Exposure and Infection Rates 
We compare the actual with the potential donor exposure and possible infection rates in the Hanover Medical School (MHH) platelet (PLT) transfusion recipients if the current MHH standard of apheresis PLT concentrate (A-PC) supply would be replaced by a pooled PLT concentrate (P-PC) transfusion regimen.
Donors, Patients, and Methods
The electronic records of the MHH Institute of Transfusion Medicine and the MHH Department of Medical Controlling were evaluated to assess the development of PLT needs and supply at MHH from 2003–2006. For 2006, we evaluated all PLT transfusion recipients with respect to their overall transfusion needs, classified them for low and high PLT transfusion needs, and related them to the diagnostic groups that underlie their PLT demands. We assumed a P-PC preparation procedure using 4 whole blood-derived buffy coats for all calculations for potential donor exposure. To predict the possible infection rates of an unrecognized viral infection with low prevalence in the general population to A-PC or to P-PC recipients and the influence of neutralizing agent specific antibodies (NAB), we established a mathematical contamination/infection model based on the current PLT transfusion mode and data about GBV-C virus infection among Hanover blood donors.
From 2003 to 2006, the 1,300–1,400 persons comprising MHH apheresis donor pool covered a 36% increase in PC transfusions. The exclusive use of P-PCs instead of A-PC would require a total of 36,240–49,276 whole blood donations to meet MHH demands, corresponding to a more than 1 log step increase in donor exposure. For individual hematological patients, the change to P-PCs would imply an 80–125%, for individual surgical patients a 40–50% higher donor exposure. Our infection model revealed an approximately 4 times higher infection.
A change to P-PC would imply a more than one log step higher donor exposure, and an unrecognized infection with a prevalence around 1% leads to an up to 4 times higher infection rate. A general change in the PC transfusion policy that favors P-PCs is dangerous and must be avoided.
PMCID: PMC3076344  PMID: 21512637
Donor exposure; Apheresis platelet concentrates; Pooled platelet concentrates; Infection rates from apheresis platelet concentrates; Infection rates from pooled platelet concentrates

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