Hideaki Matsuura

BACKGROUND AND OBJECTIVES: Reports on the changes in plasma fibrinogen levels in patients receiving cryoprecipitates synthesized using different methods are lacking. Therefore, we investigated these changes in patients who underwent cardiovascular surgery. MATERIALS AND METHODS: We included 309 patients who underwent cardiovascular surgery and received 12 cryoprecipitate units between February 2020 and March 2024 and 204 patients were selected by propensity score matching. The cryoprecipitates were prepared using two methods. Fresh frozen plasma (FFP) was thawed at 2 to 6°C for 24 h and centrifuged to remove the supernatant in the one-step method, whereas FFP was thawed, refrozen at -20°C, and subsequently rethawed in the two-step method. We investigated the association between different cryoprecipitate preparation methods and ICU admission for ≥ 1 week, with in-hospital mortality considered as a competing risk in the analysis. In addition, we evaluated the changes in plasma fibrinogen levels before and after cryoprecipitate administration. RESULTS: Baseline plasma fibrinogen levels were significantly higher in the two-step method group than in the one-step method group. Differences in cryoprecipitate preparation methods were not significantly associated with ICU admission for ≥ 1 week, in the analysis that considered in-hospital mortality as a competing risk (P = 0.93). The increase in plasma fibrinogen levels after cryoprecipitate administration was significantly higher with the two-step method than with the one-step method (36 mg/dL vs. 51 mg/dL, P = 0.020). CONCLUSION: The cryoprecipitates synthesized using the two-step method showed a higher increase in plasma fibrinogen levels than those prepared using the one-step method. These findings may help guide appropriate transfusion protocols by confirming intraoperative plasma fibrinogen levels.

This study investigated the anti-tumor effects of andrographolide, a diterpene lactone derived from Andrographis paniculata, on T-cell acute lymphoblastic leukemia (T-ALL) cells. Andrographolide induced dose-dependent cytotoxicity and morphological changes in the T-ALL cell line Jurkat cells, including cell shrinkage and chromatin condensation. Mechanistically, andrographolide triggers apoptosis through reactive oxygen species (ROS) generation, mitochondrial membrane depolarization, and cytochrome c release. These effects were reversed by the ROS inhibitor N-acetyl-L-cysteine (NAC), indicating that andrographolide induces apoptosis through a ROS-dependent apoptotic pathway. In contrast, NAC treatment did not reverse cytarabine- and vincristine-induced apoptosis or the ROS-dependent apoptotic pathway in Jurkat cells. Intriguingly, andrographolide also induced ferroptosis, as evidenced by increased expression of the ferroptosis marker fatty acid-CoA ligase 4 and ultrastructural changes such as reduced mitochondrial area and disappearance of cristae. These effects were likewise reversed by NAC, further implicating ROS in the ferroptotic process. In MOLT-4 cells, where andrographolide suppressed viability, increased Annexin V positivity and ROS levels, and upregulated FACL4 expression in a NAC-sensitive manner. Unlike cytarabine and vincristine, andrographolide did not significantly alter cell cycle distribution. In conclusion, andrographolide induces both apoptosis and ferroptosis in T-ALL cells via ROS-dependent mechanisms that are distinct from those of conventional chemotherapeutic agents. These dual actions position andrographolide as a candidate for standalone or combination therapy in T-ALL.

Abstract Background Ethylenediamine tetraacetate/glycine acid (EGA) and chloroquine diphosphate (CDP) are used in transfusion testing to dissociate IgG antibodies from red blood cells (RBCs). However, the ability of these reagents to dissociate IgM antibodies sensitized to RBCs has not been comprehensively elucidated. We investigated whether EGA and CDP could dissociate cold‐reactive antibodies from RBCs and their effect on RBCs after dissociation treatment. Study Design and Methods Cold‐reactive antibody‐sensitized RBC samples were prepared by mixing group A RBCs and group B plasma and treated with EGA, CDP, and dithiothreitol (DTT). Before and after the dissociation treatment, changes in the agglutination of these RBCs were assessed using the test tube method. Flow cytometric analysis was used to confirm the nature of antibodies bound to RBCs. Additionally, RBC morphology was evaluated using scanning electron microscopy. This study utilized off‐label use of EGA and CDP. Results Flow cytometric analysis showed that antibodies sensitized to RBCs were mainly IgM antibodies. After antibody dissociation, agglutination disappeared in the EGA‐treated samples to the same degree as in the DTT‐treated samples. However, IgM antibodies remained in the CDP‐treated samples. Regarding RBC morphology, RBC surface appeared coarser in both EGA‐ and CDP‐treated samples, and RBC area was significantly smaller in the CDP‐treated samples than in the EGA‐treated samples. Discussion EGA could dissociate cold‐reactive antibodies, whereas CDP had a higher residual antibody content. This difference in dissociation ability appears to correlate with the antibody pH of the dissociation reagent. EGA treatment may be useful in cases of sensitization by high‐titer cold‐reactive antibodies.

Abstract Background Donor-specific antibodies (DSAs) targeting human leukocyte antigens (HLAs) substantially reduce the longevity of transplanted organs. Desensitization of DSA-positive renal transplant recipients is achieved through intravenous administration of immunoglobulin (IVIg). However, the presence and detectability of anti-HLA antibodies in IVIg preparations following administration are not fully understood. We aimed to assess whether immunoglobulin preparations contain anti-HLA antibodies that can be detected as passive antibodies when administered into the body. Methods We evaluated 3 immunoglobulin preparations from different pharmaceutical companies, using anti-HLA class I and II antibody specificity tests and immunocomplex capture fluorescence analysis (ICFA). Results Direct testing for anti-HLA antibodies resulted in high background errors, particularly for Venoglobulin. Diluting Venoglobulin to physiological concentrations revealed the presence of anti-HLA class I antibodies; however, no common alleles were found between the specificity identification test and ICFA. For Glovenin and Venilon, anti-HLA class I and II antibodies were detected; however, variability was observed across different test reagent lots. Moreover, dilution of the globulin formulation revealed a prozone phenomenon. Conclusion The administration of IVIg complicates the accurate detection of anti-HLA antibodies, underscoring the need for careful interpretation of test results post-IVIg administration.

Abstract Background Despite several reports on red blood cell (RBC) alloimmunization, the actual prevalence and factors contributing to RBC alloimmunization in transfused patients remain poorly investigated. We examined the association between clinical factors and the development and evanescence of RBC antibodies after transfusion. Study Design and Methods Each participating institution performed antibody screens before and after RBC transfusion. A survey including patient characteristics, results of antibody screen and identification, antibody screen methods, total amount of RBC transfused, and adverse reactions, was conducted. Results Between October 2018 and March 2023, 1194 patients were registered at five institutions. Overall, 958 patients underwent at least one follow‐up RBC antibody screen after transfusion, revealing new antibody development in 44 (4.6%). Anti‐E was identified in 25 patients, anti‐Jk^a in 5, and anti‐c in 4. The number of RBC units transfused was significantly associated with antibody development after transfusion (p < .001). Among 55 patients in whom antibodies were identified after transfusion, including historical antibodies, antibodies evanesced in 18 (33%); anti‐E in 7, anti‐Jk^a in 4, and anti‐Le^a in 2. Evanescent antibodies were identified more frequently by saline and/or enzyme methods than persistent antibodies (p = .012). Discussion The number of RBC units transfused can impact antibody development, and antibodies identified only by saline and/or enzyme methods, deemed clinically insignificant, are likely to have a high evanescence rate. Antibody screen should be carefully performed, especially in those receiving a large number of RBC units. Confirming previous antibody screen results should be performed to prevent omitting evanesced antibodies regardless of clinical relevance.

Severe coronavirus infection (COVID-19) requires treatment with extracorporeal membrane oxygenation (ECMO). COVID-19 patients have high transfusion requirements, but there are few reports on how to manage and administer them. From 2019 to 2021, 37 patients with COVID-19 and ischemic heart disease (IHD) were enrolled. We obtained information about the patients’ background features, blood product requirements, duration of ECMO, laboratory data, and outcomes. During ECMO, transfusion was administered in both groups at the time of laboratory data reduction and ECMO weaning. IHD patients were administered each blood product in the early phase of ECMO. In contrast, COVID-19 patients were administered blood products every day during ECMO and used significantly more FFP than IHD patients. Laboratory data are generally used as a guideline for transfusion therapy, but transfusions are also administered during weaning from ECMO, it is necessary to pay attention to the status of the treatment. We need to monitor laboratory data and treatment status to manage blood products and to quickly respond to transfusion requests.

Cryoprecipitates contain high concentrations of coagulation factors such as fibrinogen. Hence, a cryoprecipitate infusion for massive bleeding may reduce blood loss and blood transfusion requirement. We introduced processes for the supply of in-hospital-prepared cryoprecipitates and found changes in transfusion-related clinical parameters in our hospital. In this study, we compared the use and non-use of cryoprecipitates in major and nonmajor cardiovascular surgeries. The parameters compared were the amounts of blood loss, red blood cells (RBC), fresh frozen plasma (FFP), and platelet concentrate administered, the ratio of RBC to FFP (R/F ratio), and the duration of intensive care unit admission. The levels of serum fibrinogen increased after cryoprecipitate infusion. However, the amount of blood loss was greater in use than non-use of cryoprecipitates for both major and nonmajor cardiovascular surgeries. There was no significant differences in RBC and FFP administered for patients use and non-use cryoprecipitates for major cardiovascular surgery. Reducing the requirement of blood products in cardiovascular surgeries requires not only the use of cryoprecipitates but also identifying the cases for which cryoprecipitate infusion is effective as well as changing the minds of the medical staff involved in transfusion therapy.

SARS-CoV-2 infection has been reported to be associated with a positive direct antiglobulin test (DAT). In this study, an analysis of 40 consecutive coronavirus disease 2019 (COVID-19) cases from December 2020 to September 2021 in Japan revealed that patients of 70 years and over were predisposed to a positive DAT. DAT positivity was related to a decrease in the hemoglobin level. Anemia in DAT-positive COVID-19 patients was attributed to hemolysis, which was corroborated by high reticulocyte counts and an increase in the red blood cell distribution width. Human leukocyte antigen (HLA)-DRB1*12:01 and DRB1*12:02 were exclusively found in DAT-positive COVID-19 patients. In silico assays for the Spike protein of SARS-CoV-2 predicted several common core peptides that met the criteria for a B cell epitope and strong binding to both HLA-DRB1*12:01 and DRB1*12:02. Among these peptides, the amino acids sequence TSNFR, which is found within the S1 subunit of SARS-CoV-2 Spike protein, is shared by human blood group antigen Rhesus (Rh) CE polypeptides. In vitro analysis showed that the expression of HLA-DR in CD4+ T cells and CD8+ T cells from a DAT-positive patient was increased after pulsation with TSNFR-sequence-containing peptides. In summary, positive DAT is related to enhanced anemia and to HLA-DR12 in the Japanese population. A peptide sequence within SARS-CoV-2 Spike protein may act as an epitope for IgG binding to RBCs in DAT-positive COVID-19 patients.

INTRODUCTION: Quantitative measurement of anti-A/-B antibody titers is important during ABO-incompatible living kidney transplantation (ABOi-LKT). METHODS: We conducted a multi-institutional study to measure the antibody titers using the automated column agglutination technique (auto-CAT) and tube test (TT) method in ABOi-LKT recipients. Statistical analysis was performed to evaluate the two methods. RESULTS: We examined 111 samples from 35 ABOi-LKT recipients at four institutions. The correlation coefficient of the two methods was >0.9; the concordance rate and clinically acceptable concordance rate for the IgG titers were 60.4% and 88.3%, respectively. Perioperative status did not influence the statistical significance. Parallel changes were observed in the IgG antibody titers measured using the auto-CAT or TT technique by desensitizing therapy in time-course monitoring. CONCLUSION: Auto-CAT is comparable with the TT technique and is feasible for IgG anti-A/B antibody titration in ABOi-LKT recipients.

A 43-year-old Japanese male, who had undergone open liver surgery for tumor resection, presented with decreased hemoglobin levels on Day 13 post-emergency-release transfusion of 16 units of Fy(a +) red blood cells. As the anemia was accompanied by increased lactate dehydrogenase, indirect bilirubin, and reticulocytes, as well as decreased haptoglobin, it was attributed to hemolysis. In the diagnostic workup for hemolytic reaction, the direct antiglobulin test result for IgG was positive and the antibody dissociated from the patient's peripheral red blood cells was identified as anti-Fya (titer, 4). The hemolytic reaction was transient (approximately 10 days), of moderate severity, and did not result in any obvious organ damage. However, a single compatible red blood cell transfusion of 2 units was required on Day 17 after the causative transfusion. Notably, HLA typing revealed that the patient carried the HLA-DRB1*04:03 allele, which has been implicated in immunogenicity and induction of anti-Fya response in Caucasian populations. In summary, this is the first documented case of definitive anti-Fya-mediated delayed hemolytic transfusion reaction associated with HLA-DRB1*04:03 in the Japanese population.

BACKGROUND: The occurrence of transfusion-transmitted hepatitis B virus (HBV) infection has fallen dramatically due to continuous improvements in pre-transfusion laboratory testing. However, the characteristics of transfusion-transmitted HBV infection caused by individual donor nucleic acid amplification test (ID-NAT)-negative blood products are unclear. CASE PRESENTATION: A 76-year-old woman with acute myeloid leukemia was diagnosed with transfusion-transmitted HBV infection after receiving apheresis platelets derived from an ID-NAT-negative blood donation. This case was diagnosed definitively as transfusion-mediated because complete nucleotide homology of a 1556 bp region of the HBV Pol/preS1-preS2-S genes and a 23 bp region of the HBV core promoter/precore between the donor and recipient strains was confirmed by PCR-directed sequencing. The case is uncommon with respect to the unexpectedly prolonged HBV-DNA incubation period of nearly 5 months after transfusion (previously, the longest period observed since the recent implementation of ID-NAT pre-transfusion laboratory testing in Japan was 84 days). Slow-replicating HBV genotype A2 may contribute to the prolonged incubation period; also, the quantity of apheresis platelets delivered in a large volume of plasma, and/or the immune response of the recipient suffering from a hematological neoplasm, may have contributed to establishment of HBV infection in the recipient. This was supported by analysis of three previously documented cases of transfusion-transmitted HBV infection by blood products derived from ID-NAT-negative donations in Japan. CONCLUSION: Continuous monitoring of HBV infection for longer periods (>3 months) may be required after transfusion of blood components from an ID-NAT-negative HBV window donation.

OBJECTIVE: The JT interval of the myocardial repolarization time can be divided into Jpoint to T-peak interval (JTp) and T-peak to T-end interval (Tpe). It is well known that the JT interval is dependent on the heart rate, but little is known regarding heart rate dependence for JTp and Tpe. The aim of the present study was to clarify the heart rate dependence of JTp and Tpe and to elucidate the interference of autonomic nervous activity with these parameters. METHODS: We evaluated 50 prepubertal children (mean age: 6.4 ± 0.5 years; male:female, 22:28) without heart disease. JTp, Tpe, and the preceding RR intervals were measured using 120 consecutive beats (lead CM5). First, the relationships between the RR interval and JTp and Tpe were evaluated by Pearson's correlation coefficient. Second, to evaluate autonomic interference with JTp and Tpe, the degree of coherence between RR interval variability and JTp or Tpe variability was calculated using spectral analysis. RESULTS: Significant positive correlations were observed between the RR interval and JTp (y = 0.116x + 105.5; r = 0.594, p < 0.001) and between the RR interval and Tpe (y = 0.037x + 44.7; r = 0.432, p < 0.001). Tpe variability had a lower degree of coherence with RR interval variability (range: 0.039-0.5 Hz) than with JTp variability (0.401 [interquartile range, 0.352-0.460] vs. 0.593 [0.503-0.664], respectively; p < 0.001). CONCLUSIONS: Tpe had lower heart rate dependence and a lower degree of autonomic nervous interference than did JTp.

輸血チーム医療の現状を把握するために行ったアンケート調査を踏まえて、輸血チーム医療を推進していくための方略について検討した。アンケートは医療スタッフ164名のうち142名(86.6%)から回答が得られた。アンケート結果からは「他職種との相互理解不足」が輸血医療チームの推進を妨げる主な要因であることが示唆された。また、職種によって現状のチーム医療に対する認識は異なることが明らかになった。以上、今回の調査結果から各施設においてコミュニケーション不足や相互理解不足によりミスマッチが生じやすい緊急輸血等の場面を想定したシミュレーションを実施し、輸血医療チームにおける他職種との相互理解や他職種の協力を得る努力を行っていく必要があると考えられた。

BACKGROUND AND OBJECTIVES: Determination of the anti-A/-B titre pre- and post-transplantation is beneficial for treatment selection. Currently, the recommended method for antibody titration is the tube test (TT) assay. Dithiothreitol (DTT) is used for IgM antibody inactivation. Recently, a fully automated antibody titration assay using the column agglutination technique (CAT) was developed (auto-CAT). Our aim was to compare the auto-CAT and TT techniques for ABO antibody titration, to evaluate the effectiveness of DTT-treated plasma for use with auto-CAT and to define the cut-off value for antibody titration by auto-CAT. MATERIALS AND METHODS: We enrolled 30 healthy individuals, including 10 each for blood types A, B and O. We performed antibody titre measurement using the TT technique and auto-CAT simultaneously. Auto-CAT uses the bead column agglutination technology. RESULTS: With the auto-CAT cut-off value set to weak (w)+ with DTT treatment plasma, the concordance rate was 45%, and the weighted kappa value between TT and auto-CAT results was 0·994 in all subjects. Furthermore, there was a significant positive correlation between the anti-A/-B titre results obtained using the TT technique and auto-CAT in all blood types. Moreover, a positive bias (falsely elevated end-points due to agglomeration of A/B cells) was not observed in auto-CAT testing using DTT-treated plasma. CONCLUSION: Our results show that 1+ agglutination using the TT technique is equivalent to w+ agglutination obtained using auto-CAT. We recommend that DTT may be used with auto-CAT to measure antibody titres. Thus, we suggest that auto-CAT is useful for antibody titration in routine examination.

&lt;B&gt;Purpose&lt;/B&gt;: Left atrial (LA) volume index (LAVI) has been reported to reflect left ventricular (LV) diastolic dysfunction and predict cardiac events in patients with coronary artery disease (CAD). Late diastolic mitral annular velocity (A&amp;rsquo;) has been shown to correlate well with LA function and LV filling pressure. We hypothesized that A&amp;rsquo; could be a useful predictor of mortality and rehospitalization due to heart failure (HF) in CAD patients with dilated LA volume. &lt;B&gt;Subjects and Methods&lt;/B&gt;: This study included 212 patients (mean age: 64 years, 166 men) who were admitted to the coronary care unit at our hospital due to acute coronary syndrome (ACS). They underwent echocardiography including tissue Doppler imaging (TDI). None of them had atrial fibrillation and atrial flutter or more than moderate mitral valvular disease. The primary study endpoints were mortality and rehospitalization due to HF. &lt;B&gt;Results and Discussion&lt;/B&gt;: During a mean follow-up of 508 days, eight patients died and nine patients were rehospitalized due to HF. According to a previous report, the patients were divided into two groups. Group A: LAVI&amp;ge;32 ml/m&lt;SUP&gt;2&lt;/SUP&gt; (n=62) and Group B: LAVI&amp;lang;32 ml/m&lt;SUP&gt;2&lt;/SUP&gt; (n=150). Receiver operator curve analysis revealed that A&amp;rsquo;&amp;ge;10.7 cm/s was the optimal cut-off value to predict mortality and rehospitalization due to HF in Group A. In Group A, patients with A&amp;rsquo;&amp;ge;10.7 cm/sec showed a significantly lower incidence of cardiac events than those with A&amp;rsquo;&amp;lang;10.7 cm/sec (log-rank, p=0.0019). However, A&amp;rsquo; could not be a predictor of events in Group B. &lt;B&gt;Conclusion&lt;/B&gt;: A&amp;rsquo; could be a useful predictor of mortality and rehospitalization due to HF in ACS patients with large LAVI.

The relationship between electrocardiographic RR and QT interval variabilities were evaluated, based on the QT variability ratio (VR) in prepubescent children, to investigate the influence of aging. One hundred and seventy-six subjects with no heart disease, ranging from infants to school children, were categorized into 6 groups for a comparative study ; Group I(17 infants aged 0 to 6 months), Group II(21 infants aged 7 to 11 months), Group III(28 infants aged 1 year), Group IV(37 children aged 2 to 3 years), Group V(36 children aged 4 to 5 years), Group VI(37 children aged 6 to 7 years). Regarding the methods, the RR-interval and QT-interval of 120 heart beats were measured at rest to calculate the VR based on the standard deviation of the RR-interval (SDRR) and the standard deviation of the QT-interval (SDQT) in order to investigate the relationship with the age of subjects (months). As a result, there were no gender differences between groups for VR, SDRR and SDQT, and VR decreased in accordance with aging, but became stable when the children reached school age. This study supports the potential application of the standard value of VR by age to evaluate imbalances in the cardiac cycle and myocardial repolarization.

&lt;B&gt;Background&lt;/B&gt;: The ratio of early transmitral velocity to early diastolic velocity of the mitral annulus (E/E&amp;prime;) has been shown to be an excellent predictor of left ventricular filling pressure. However, the clinical significance of mildly elevated E/E&amp;prime; (8&amp;le;E/E&amp;prime;&amp;le;15) remains to be clarified. Left atrial (LA) volume would reflect the duration and severity of left ventricular diastolic dysfunction. There is a graded relation between LA volume index (LAVI) and severity of diastolic dysfunction. &lt;B&gt;Purpose&lt;/B&gt;: We hypothesized that LAVI could be useful for predicting increased PAWP in coronary heart disease patients with mildly elevated E/E&amp;prime;. &lt;B&gt;Subjects and Methods&lt;/B&gt;: Patients admitted to the coronary care unit at our hospital who had indwelling pulmonary artery catheters were eligible. Fifty-eight patients without atrial fibrillation or severe mitral regurgitation underwent echocardiography (Sonos 5500 or 7500; S3 probe; Philips). Tissue Doppler imaging of the mitral annulus was also obtained. Early diastolic E&amp;prime; velocity was measured from the septal mitral annulus velocity profile in the apical 4-chamber view. LA volume was assessed by the biplane Simpson&amp;prime;s method from apical 4- and 2-chamber views in end systole. LAVI was obtained by correcting for body surface area. PAWP was measured simultaneously in all the patients. &lt;B&gt;Results&lt;/B&gt;: In 23 patients with mildly elevated E/E&amp;prime; (8&amp;le;E/E&amp;prime;&amp;le;15) , there was no correlation between PAWP and E/E&amp;prime; (p=0.40). However, LAVI positively correlated with PAWP in those patients (r=0.64, p&amp;lang;0.001). LAVI&amp;ge;32ml/m&lt;SUP&gt;2&lt;/SUP&gt; was the optimal cutoff to predict PAWP&amp;ge;15mmHg (sensitivity 72%, specificity 80%). &lt;B&gt;Conclusion&lt;/B&gt;: LAVI could be a useful adjunct measure for predicting increased PAWP in coronary heart disease patients with mildly elevated E/E&amp;prime;.

早産低出生体重児13例を対象に,脳波,眼球運動電位図,心電図,呼吸波を記録・解析し,日齢0と生後1ヵ月で比較した.日齢0から生後1ヵ月にかけて洞結節興奮周期(PP間隔)は有意な短縮を示し,房室結節伝導時間(PR間隔)は延長した.周波数特性をみると,PP,PRともに低周波数領域(LF)と高周波数領域(HF)の成分が増加し,呼吸周波数領域(RSA)の成分も増加していた.PP間隔の周波数解析で,LF/HFは日齢0の7.56から生後1ヵ月には3.28へと有意に低下し,RSA/全周波数領域(TF)は0.015から0.048に有意に増加していた.生理的な心周期に対する自律神経活動の増加と,呼吸性迷走神経活動の発現が示唆された.PRではLF/HFが0.86から1.37へと有意に増加したが,RSA/TFは0.138から0.125と有意な変化はなかった.房室伝導に対する交感神経活動は亢進していたが,呼吸迷走神経入力は微弱であり,房室伝導は呼吸以外の迷走神経系により優位に支配されていることが示唆された