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  • Wiley  (13)
  • Satake, Masahiro  (13)
  • 1
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    Refined methods to evaluate the in vivo hemostatic function and viability of transfused human platelets in rabbit models
    Wiley ; 2017
    In:  Transfusion Vol. 57, No. 8 ( 2017-08), p. 2035-2044
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    In: Transfusion, Wiley, Vol. 57, No. 8 ( 2017-08), p. 2035-2044
    Abstract: To bridge the gap between in vitro function and clinical efficacy of platelet (PLT) transfusion products, reliable in vivo PLT functional assays for hemostasis and survival in animal models are required. However, there are no standardized methods for assessing the in vivo quality of transfused human PLTs. STUDY DESIGN AND METHODS Plasma‐depleted human PLT concentrates (PCs; Day 3, Day 5, Day 7, Day 10, and damaged) were transfused into busulfan‐induced rabbits with thrombocytopenia with prolonged bleeding times 1 day after treatment with ethyl palmitate (EP) to block their reticuloendothelial systems. The hemostatic effect of PC transfusion was evaluated by the ear fine vein bleeding time. For the in vivo survival assay, splenectomized EP‐treated rabbits were transfused with human PCs, and viability of the human PLTs in the rabbits was determined by flow cytometry using human PLT‐specific antibodies and Trucount tubes. RESULTS The hemostatic effect of PCs was slightly reduced with increasing storage periods for early time points, but more dramatically reduced for later time points. PLT survival was similar after 3 and 7 days of storage, but PLTs stored for 10 days showed significantly poorer survival than those stored only 3 days. CONCLUSION Our new and improved protocol for in vivo assessment of transfused PLTs is sufficiently sensitive to detect subtle changes in hemostatic function and viability of human PLTs transfused into rabbit models. This protocol could contribute to preclinical in vivo functional assessment and clinical quality assurance of emerging novel PLT products such as cultured cell–derived human PLTs.
    Type of Medium: Online Resource
    ISSN: 0041-1132 , 1537-2995
    URL: Issue
    URL: Article
    DOI: 10.1111/trf.2017.57.issue-8
    DOI: 10.1111/trf.14189
    Language: English
    Publisher: Wiley
    Publication Date: 2017
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  • 2
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    In vitro thrombus formation and in vivo hemostasis mediated by platelets irradiated with bactericidal ultraviolet C from xenon flash under flow conditions
    Wiley ; 2021
    In:  Transfusion Vol. 61, No. 1 ( 2021-01), p. 191-201
    Details
    In: Transfusion, Wiley, Vol. 61, No. 1 ( 2021-01), p. 191-201
    Abstract: We previously reported a flow path–ultraviolet C (UVC) irradiation system for platelet concentrates (PCs) with platelet additive solution (PAS) to minimize contamination by bacteria. Here, we investigated functionalities of irradiated platelets (PLTs) in in vitro thrombus formation and in vivo hemostasis. Study Design and Methods PAS‐PCs were irradiated with flash UVC using the flow path system. Their variables (PLT count, mean platelet volume, pH, glucose, lactate, glycoprotein [GP] Ib, and activated integrin αIIbβ3) were evaluated. Static adhesion to collagen or fibrinogen was analyzed using fluorescent microscopy. Thrombus formation under flow conditions was assessed using a collagen‐coated bead column. Adenosine diphosphate (ADP)‐induced Akt phosphorylation was determined by western blot. In vivo hemostasis and circulatory survival of PLTs were assessed with a rabbit bleeding model. Results All variables, except for GPIb expression, were slightly, but significantly, impaired after flash UVC irradiation throughout the 6‐day storage period. No difference was observed in static adhesion to either collagen or fibrinogen between irradiated and nonirradiated PAS‐PCs. In vitro thrombus formation of flash UVC‐irradiated PAS‐PCs was significantly greater than that of nonirradiated PAS‐PCs. ADP‐induced Akt phosphorylation was enhanced in irradiated PAS‐PCs. In vivo hemostatic efficacy was comparable between the groups on Day 1. The efficacy declined in nonirradiated PAS‐PCs on Day 5, while it was retained in flash UVC‐irradiated PAS‐PCs. Circulatory survival of PLTs was lower in irradiated PAS‐PCs. Conclusions PAS‐PCs irradiated with UVC from xenon flash have favorable properties to achieve hemostasis compared with nonirradiated PAS‐PCs.
    Type of Medium: Online Resource
    ISSN: 0041-1132 , 1537-2995
    URL: Issue
    URL: Article
    DOI: 10.1111/trf.v61.1
    DOI: 10.1111/trf.16138
    Language: English
    Publisher: Wiley
    Publication Date: 2021
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  • 3
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    Hemostatic function of cold‐stored platelets in a thrombocytopenic rabbit bleeding model
    Wiley ; 2022
    In:  Transfusion Vol. 62, No. 11 ( 2022-11), p. 2304-2313
    Details
    In: Transfusion, Wiley, Vol. 62, No. 11 ( 2022-11), p. 2304-2313
    Abstract: Transfusion of cold‐stored platelet concentrates (CS‐PCs) appears effective in massively bleeding patients. However, few studies have evaluated their in vivo hemostatic function in severe thrombocytopenia. Study Design and Methods The in vivo function of plasma‐depleted human PCs was evaluated in rabbits with a blocked reticuloendothelial system and busulfan‐induced thrombocytopenia. On day 1, a human apheresis PC was processed in a platelet additive solution (PAS‐PC) and split evenly for cold or room temperature storage (RTS). On days 3, 6, or 9, RTS‐ or CS‐PAS‐PCs were transfused (4.0 × 10 9 platelets/kg) after plasma depletion into two to four rabbits that developed adequate thrombocytopenia ( 〈 25 × 10 9 /L). Ear bleeding time was measured by two incisions in small veins. The hemostatic rate was defined as the percentage of rabbits achieving bleeding cessation within 600 s at either incision. The experiment was repeated using five different PCs on each storage day. Results The mean pre‐transfusion rabbit platelet count was 8.6 ± 5.2 × 10 9 /L. The hemostatic rates with RTS‐ and CS‐PAS‐PCs were both 100% on day 3, 93 ± 15% and 73 ± 15% on day 6 ( p  = .07), and 65 ± 36% and 73 ± 37% on day 9 ( p  = .27), respectively, with no statistical differences. Total platelet counts were significantly lower after CS‐PAS‐PC than RTS‐PAS‐PC transfusion on all days (e.g., 58.7 ± 5.7 vs. 42.4 ± 14.7 × 10 9 /L, p  = .0007, day 9), and did not reach 50 × 10 9 /L in several experiments. Platelet count increments correlated significantly with hemostatic efficacy for CS‐PAS‐PC transfusion only. Discussion CS‐PAS‐PCs might achieve similar hemostasis as RTS‐PAS‐PCs in thrombocytopenic patients with mild bleeding. Hemostatic efficacy could be improved by transfusing more CS‐PAS‐PCs.
    Type of Medium: Online Resource
    ISSN: 0041-1132 , 1537-2995
    URL: Issue
    URL: Article
    DOI: 10.1111/trf.v62.11
    DOI: 10.1111/trf.17128
    Language: English
    Publisher: Wiley
    Publication Date: 2022
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  • 4
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    Reduction of bacteria and human immunodeficiency virus Type 1 infectivity of platelet suspension in plasma using xenon flash‐pulse light in a bench‐scale trial
    Wiley ; 2016
    In:  Transfusion Vol. 56, No. 9 ( 2016-09), p. 2256-2266
    Details
    In: Transfusion, Wiley, Vol. 56, No. 9 ( 2016-09), p. 2256-2266
    Abstract: Current pathogen reduction systems for platelet concentrates (PCs) require addition of chemical compounds and/or reduction of plasma content in PCs. We have investigated a new method using xenon (Xe) flash‐pulse light without additional compounds or plasma replacement. STUDY DESIGN AND METHODS An aliquot of apheresis platelets (PLTs) in plasma inoculated with bacteria or human immunodeficiency virus Type 1 (HIV‐1) was irradiated with Xe flash‐pulse light (Xe flash phototreatment). Bacterial growth was monitored up to 6 days of storage, whereas HIV‐1 infectivity was assayed just after treatment. Pairs of Xe flash‐phototreated and untreated PCs were examined for PLT lesion during the storage period. RESULTS Under the current conditions, a low titer (1.8 colony‐forming units [CFUs]/mL) of Staphylococcus aureus did not proliferate during the 6‐day storage period, but grew in some cases at high‐titer (24.0 CFUs/mL) inoculation. HIV‐1 infectivity was reduced by 1.8 log. PLT recovery of the treated PCs was lower than untreated ones. An increase of mean PLT volume and glucose consumption, together with a decrease of hypotonic shock response and pH, were enhanced by the treatment. CD62P‐ and PAC‐1–positive PLTs increased after the treatment, indicating the induction of PLT activation. Among biologic response modifiers, soluble CD40 ligand was significantly increased in the treated PCs on Day 6. CONCLUSIONS Xe flash phototreatment could prevent bacterial proliferation and reduce HIV‐1 infectivity in 100% plasma PCs without any additional compounds, but enhanced PLT storage lesions. Further improvement is required to increase the potency of pathogen inactivation with reducing PLT damage.
    Type of Medium: Online Resource
    ISSN: 0041-1132 , 1537-2995
    URL: Issue
    URL: Article
    DOI: 10.1111/trf.2016.56.issue-9
    DOI: 10.1111/trf.13685
    Language: English
    Publisher: Wiley
    Publication Date: 2016
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  • 5
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    Application of immortalized human erythroid progenitor cell line in serologic tests to detect red blood cell alloantibodies
    Wiley ; 2018
    In:  Transfusion Vol. 58, No. 11 ( 2018-11), p. 2675-2682
    Details
    In: Transfusion, Wiley, Vol. 58, No. 11 ( 2018-11), p. 2675-2682
    Abstract: Antibody screening in pretransfusion tests is necessary to avoid critical complications of blood transfusion. Although red blood cells (RBCs) expressing relevant alloantigen(s) have been used for serologic antibody screening, little attention has been given to the use of cell lines, in which blood group antigen gene(s) are transduced, as reagent RBCs for antibody screening. STUDY DESIGN AND METHODS The use of an erythroid progenitor cell line for serologic tests was studied. The expression of blood group antigens of erythroid progenitor cells was analyzed by genotyping and flow cytometry. Serologic analysis including hemagglutination was performed using erythroid progenitor cells to evaluate their sensitivity for antibody detection. Overexpression of exogenous erythroid antigen by lentiviral transduction was carried out and investigated for antibody detection sensitivity. RESULTS Erythroid progenitor cells contained a substantial amount of hemoglobin and expressed sufficient levels of blood group antigens to detect corresponding monoclonal antibodies. Furthermore, the cell line could acquire an exogenous RBC antigen after lentiviral transduction and detected corresponding monoclonal and alloantibodies with equal sensitivity to antigen‐positive RBCs. CONCLUSION Application of erythroid progenitor cell lines for screening for unexpected antibodies could be helpful in solving issues such as reagent availability associated with the conventional RBC‐based assay. The genetic expandability of erythroid progenitor cell lines by gene modification techniques could lead to the development of more convenient reagent RBCs.
    Type of Medium: Online Resource
    ISSN: 0041-1132 , 1537-2995
    URL: Issue
    URL: Article
    DOI: 10.1111/trf.2018.58.issue-11
    DOI: 10.1111/trf.14840
    Language: English
    Publisher: Wiley
    Publication Date: 2018
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  • 6
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    Real‐time measurement of platelet shape change by light scattering under riboflavin and ultraviolet light treatment
    Wiley ; 2016
    In:  Transfusion Vol. 56, No. 3 ( 2016-03), p. 587-595
    Details
    In: Transfusion, Wiley, Vol. 56, No. 3 ( 2016-03), p. 587-595
    Abstract: The adoption of pathogen reduction technologies (PRTs) is considered for the implementation of safer platelet (PLT) transfusion. However, the effects of PRT treatment including irradiation with ultraviolet (UV) light on PLT shape have not yet been fully clarified. STUDY DESIGN AND METHODS Leukoreduced PLT concentrates (PCs) were treated with riboflavin and UV light (Mirasol PRT, TerumoBCT). PLT shape and adenosine diphosphate (ADP)‐induced shape change were evaluated by a light scattering method where the amplitude of the scattered signal intensity was measured as the indicator of the proportion of discoid PLTs. Using a modified fluorometer, the real‐time effects of different wavelengths of UV light on PLT shape were examined over the range of 300 to 360 nm at the same dose. RESULTS The proportion of discoid PLTs in the Mirasol PRT–treated PCs decreased immediately after treatment. The difference in the proportion between PRT‐treated and untreated PLTs became larger with storage. Although this modification correlated significantly with the pH decrease and P‐selectin expression, the Mirasol PRT–treated PLTs retained sufficient ability to undergo an ADP‐induced shape change. In the study using the modified fluorometer, the proportion of discoid PLTs significantly decreased with the wavelength ( 〈 320 nm) of irradiated UV light. CONCLUSION Mirasol PRT treatment of PCs decreases the proportion of discoid PLTs, which seemed to be caused by the irradiation with UV light of short wavelengths ( 〈 320 nm), not that of long wavelengths (≥320 nm) in the Mirasol PRT system. Modification of UV light wavelength may improve the quality of PRT‐treated PCs.
    Type of Medium: Online Resource
    ISSN: 0041-1132 , 1537-2995
    URL: Issue
    URL: Article
    DOI: 10.1111/trf.2016.56.issue-3
    DOI: 10.1111/trf.13404
    Language: English
    Publisher: Wiley
    Publication Date: 2016
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  • 7
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    Analysis of antigen‐antibody cross‐reactivity among lineages and sublineages of Babesia microti parasites using human babesiosis specimens
    Wiley ; 2018
    In:  Transfusion Vol. 58, No. 5 ( 2018-05), p. 1234-1244
    Details
    In: Transfusion, Wiley, Vol. 58, No. 5 ( 2018-05), p. 1234-1244
    Abstract: Human babesiosis is caused mainly by Babesia microti and has recently become a public health concern due to an increase in transfusion‐transmitted infection. Thus, the development of an antibody detection method with high specificity and sensitivity is a priority. Seroreactivity against B. microti has been reported to be highly specific not only to B. microti lineages but also to sublineages. This study aimed to elucidate the human antibody reactivity against various lineages, including US, Kobe, and Hobetsu, and sublineages (North America and East Asia) in the US lineage. STUDY DESIGN AND METHODS Twenty samples obtained from individuals infected with B. microti in the United States were tested for the presence of anti‐ B. microti antibodies using indirect immunofluorescence assay (IFA) and Western blotting (WB) to indicate antigens of each (sub‐)lineage. RESULTS By IFA, 20 samples showed reactivity to the North America sublineage (titer range, 64‐4096), 16 to the East Asia sublineage (64‐512), 10 to the Kobe (64‐128), and five to the Hobetsu (64). Antibody titers to the East Asia sublineage, Kobe, and Hobetsu were significantly lower than those to the North America sublineage (p  〈  0.01). By WB, in parallel with the IFA results, 18 samples showed strong reactions to the North America sublineage, weak reactions to the East Asia sublineage, and near‐zero reactions to the Kobe and Hobetsu. CONCLUSION Human antibodies induced by B. microti infection are highly specific against B. microti lineages and sublineages with low cross‐reactivity. Developing a precise antibody detection method may require specific antigens based on B. microti lineages and sublineages.
    Type of Medium: Online Resource
    ISSN: 0041-1132 , 1537-2995
    URL: Issue
    URL: Article
    DOI: 10.1111/trf.2018.58.issue-5
    DOI: 10.1111/trf.14558
    Language: English
    Publisher: Wiley
    Publication Date: 2018
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  • 8
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    Development of a novel dengue virus serotype‐specific multiplex real‐time reverse transcription–polymerase chain reaction assay for blood screening
    Wiley ; 2016
    In:  Transfusion Vol. 56, No. 12 ( 2016-12), p. 3094-3100
    Details
    In: Transfusion, Wiley, Vol. 56, No. 12 ( 2016-12), p. 3094-3100
    Abstract: Dengue fever is caused by four related RNA viruses of the genus Flavivirus , dengue virus (DENV)‐1, ‐2, ‐3, and ‐4, which are transmitted to humans by mosquitoes. Although DENV is not endemic in Japan, an autochthonous dengue outbreak occurred in 2014. Several transfusion‐transmitted cases have also been reported after the use of blood and plasma products in DENV‐endemic countries. The aim of this study was to develop a novel multiplex reverse transcription–polymerase chain reaction (RT‐PCR) assay for DENV blood screening. STUDY DESIGN AND METHODS Large‐scale oligonucleotide screening was performed to obtain DENV‐specific primers and probes using a variety of DENV clinical isolates. A multiplex RT‐PCR assay was then developed using the identified oligonucleotides and the ability of this assay to detect DENV RNA was evaluated. RESULTS A number of oligonucleotides suitable for DENV RNA detection were identified and a novel DENV serotype‐specific multiplex RT‐PCR assay was successfully established. Comparative analysis revealed that the multiplex assay could detect levels of viral contamination as low as 100 viral copies/mL. CONCLUSION This established serotype‐specific multiplex RT‐PCR assay provides a simple, sensitive, and quantitative detection method for DENV, which could be applied in the screening of blood samples to prevent transfusion‐transmitted DENV infection.
    Type of Medium: Online Resource
    ISSN: 0041-1132 , 1537-2995
    URL: Issue
    URL: Article
    DOI: 10.1111/trf.2016.56.issue-12
    DOI: 10.1111/trf.13875
    Language: English
    Publisher: Wiley
    Publication Date: 2016
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  • 9
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    Effects of riboflavin and ultraviolet light treatment on platelet thrombus formation and thrombus stability on collagen
    Wiley ; 2017
    In:  Transfusion Vol. 57, No. 7 ( 2017-07), p. 1772-1780
    Details
    In: Transfusion, Wiley, Vol. 57, No. 7 ( 2017-07), p. 1772-1780
    Abstract: Pathogen reduction technologies (PRTs) are considered for the implementation of safer platelet (PLT) transfusion. PRT treatment involves the addition of a photosensitizer to a blood component followed by ultraviolet (UV) light irradiation. However, the effects of PRT treatment on PLT thrombus formation and thrombus stability have not been satisfactorily clarified. STUDY DESIGN AND METHODS Leukoreduced PLT concentrates (PCs) were treated with riboflavin and UV light (Mirasol PRT). PLT thrombus formation on collagen was evaluated by the microchannel method, by which the total amount of PLTs deposited was measured as indices of thrombus formation and thrombus stability. Using a cone‐plate shear‐induced PLT aggregometer, PLT reactivity in blood flow was examined in a wide range of shear stresses of 6 to 108 dyn/cm 2 . RESULTS There was no significant difference in surface coverage between PRT‐treated PLTs and control PLTs on collagen. On the other hand, the total amount of PRT‐treated PLTs deposited was higher than that of control PLTs. The promotive effect of PRT treatment on PLT deposition completely disappeared in the presence of tirofiban, a potent integrin αIIbβ3 inhibitor. The percentage of the dissociation of PRT‐treated PLTs on collagen was lower than that of control PLTs after flushing with phosphate‐buffered saline. PRT treatment significantly inhibited PLT aggregation under high‐shear‐stress conditions. CONCLUSION Riboflavin‐based PRT treatment of PCs leads to the enhancement of PLT thrombus formation and thrombus stability on collagen. However, it does not enhance the reactivity of PLTs not in contact with collagen under high‐shear‐stress conditions.
    Type of Medium: Online Resource
    ISSN: 0041-1132 , 1537-2995
    URL: Issue
    URL: Article
    DOI: 10.1111/trf.2017.57.issue-7
    DOI: 10.1111/trf.14114
    Language: English
    Publisher: Wiley
    Publication Date: 2017
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  • 10
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    Restored response to ADP downstream of purinergic P2Y 12 receptor in apheresis platelets after pathogen‐reducing xenon flash treatment
    Wiley ; 2018
    In:  Transfusion Vol. 58, No. 5 ( 2018-05), p. 1117-1125
    Details
    In: Transfusion, Wiley, Vol. 58, No. 5 ( 2018-05), p. 1117-1125
    Abstract: Our previous study revealed that pathogen‐reducing filtered xenon flash‐treated platelets (fXe‐PLTs) showed sustained aggregation in response to adenosine diphosphate (ADP), but apheresis‐collected PLTs (Aph‐PLTs) showed reversible aggregation. STUDY DESIGN AND METHODS Aph‐PLTs, fXe‐PLTs, and freshly prepared PLTs (PRP‐PLTs) from whole blood were used to investigate the following responses to ADP: concentration response and effects of ADP receptor antagonists on aggregation, the cytosolic calcium (Ca 2+ ) flux downstream of P2Y 1 receptor signaling, and phosphorylation of vasodilator‐stimulated phosphoprotein (VASP) and signaling intermediate protein Akt downstream of the P2Y 12 receptor. RESULTS The aggregation of Aph‐PLTs by ADP (10 µM) changed from reversible to sustained in an fXe flash dose‐dependent manner. The concentration‐response curve of Aph‐PLTs showed a fivefold higher 50% effective concentration compared with PRP‐PLTs, and fXe treatment decreased it to threefold. While the basal Ca 2+ level was higher both in Aph‐ and fXe‐PLTs than in PRP‐PLTs, the increase of cytosolic Ca 2+ by ADP remained unchanged in Aph‐ and PRP‐PLTs, but was slightly reduced in fXe‐PLTs. Although the forskolin‐induced VASP phosphorylation was significantly reduced in Aph‐PLTs, and partially restored by the fXe treatment, ADP stimulation attenuated this phosphorylation to an equivalent extent among the three PLT types. The ADP‐stimulated time‐dependent Akt phosphorylation was weak in Aph‐PLTs, whereas fXe‐PLTs and PRP‐PLTs showed a marked increase. CONCLUSION These results indicate that the reversible aggregation of Aph‐PLTs is the consequence of insufficient Akt phosphorylation. The fXe treatment restores the increase of phosphorylated Akt, resulting in the sustained aggregation of fXe‐PLTs similar to those of PRP‐PLTs.
    Type of Medium: Online Resource
    ISSN: 0041-1132 , 1537-2995
    URL: Issue
    URL: Article
    DOI: 10.1111/trf.2018.58.issue-5
    DOI: 10.1111/trf.14578
    Language: English
    Publisher: Wiley
    Publication Date: 2018
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