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BACKGROUND Coronavirus disease 2019 (COVID-19) testing is essential, as it enables early detection and halting the spread of infection throughout the community. Real-time reverse-transcriptase polymerase chain reaction (real-time RT-PCR) testing is the predominant method for detecting severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in samples, and the cycle threshold value (Ct value) is used to determine COVID-19 positivity. OBJECTIVE The aim of the present study was to examine time series distribution during the COVID-19 pandemic, using Ct values at the national level in Korea, and analyze the association between Ct values and the time-varying reproduction number (Rt) to discuss the utilization of laboratory-based COVID-19 test results. METHODS We used real-time RT-PCR results collected by the Seegene Medical Foundation from the index case in Korea between February 2020 and January 2022. The distribution of daily Ct values (RdRp/S target) was examined and compared with the daily count of newly diagnosed cases and Rt to determine the usability of Ct values. Moreover, time lag was applied to the daily count of newly diagnosed cases to analyze the association between Ct values and Rt. RESULTS During the COVID-19 pandemic, Ct values declined in general, while viral load increased progressively. After Ct values dropped markedly, the number of newly diagnosed cases rose substantially, and the association analysis confirmed that the daily count of newly diagnosed cases declined with increasing Ct values. The time series trend of the Ct values was also similar to that of Rt, a classic marker that was used as a predictor of pandemic trends; when compared to the actual count of newly diagnosed cases, we found that Ct values could be used to predict new diagnoses earlier than Rt. The fact that the Ct values were more sensitive to a substantial rise of new COVID-19 cases than Rt in the early days of the pandemic also supports this finding. CONCLUSIONS We examined the potential of Ct values as a predictor of new COVID-19 cases in real-time using nationally collected Ct value data. Further, we proposed the use of Ct values as an index reflecting the degree of viral load. The findings of this study can be used to establish indicators with higher predictive power than conventional outbreak predictions.

During the coronavirus disease 2019 (COVID-19) pandemic, COVID-19 testing is crucial, as it enables early detection and halting the spread of infection throughout the community. Real-time reverse-transcriptase polymerase chain reaction (Real-time RT-PCR) testing is the predominant method for COVID-19 testing, and the cycle threshold value (Ct value) is used to determine COVID-19 positivity. There are many ongoing studies using Ct value, and the present study aims to examine time series distribution during the pandemic using Ct values at the national level and analyze the association with time-varying reproduction number (Rt) to discuss the utilization of laboratory-based COVID-19 test results. We used Real-time RT-PC results collected by Seegene Medical Foundation from the index case in Korea in February 2020 to January 2022 in Korea. The distribution of daily Ct value ( RdRp/S target) was examined, and it was compared with the daily count of newly diagnosed cases and Rt to determine the usability of Ct values. Moreover, time lag was applied to the daily count of newly diagnosed cases to analyze the association between Ct values and Rt. During the COVID-19 pandemic, Ct values declined in general, while viral load increased progressively. After Ct values dropped markedly, the number of newly diagnosed cases rose substantially, and the association analysis also confirmed that the daily count of newly diagnosed cases declined with increasing Ct values. The time series trend of the Ct values was also similar to that of Rt, a classic marker used as a predictor of the trends of the pandemic, and when compared to the actual count of newly diagnosed cases, Ct values can be used to predict new diagnoses earlier than Rt. The fact that the Ct values were more sensitive to a substantial rise of new COVID-19 cases than Rt was in the early days of the pandemic also support this. We examined the potential of Ct values as a predictor of new COVID-19 cases in real-time using nationally collected Ct value data. Further, we proposed the use of Ct values as an index reflecting the degree of viral load, so the findings of this study can be used as valuable evidence to support public health decisions for response and resource distribution.

Background: DEL, the weakest D variant, is mistyped as D-negative by routine serological assays. Transfusion of red blood cells expressing the DEL phenotype has the potential to elicit anti-D alloimmunization in D-negative recipients. The goal of this study was to recommend DEL typing strategies for serologically D-negative Asian donors. Methods: RhCE phenotyping and the adsorption-elution test were performed on 674 serologically D-negative samples. RHD genotyping using real-time polymerase chain reaction and melting curve analysis were also undertaken to identify DEL alleles. Costs and turnaround time of RhCE phenotyping, the adsorption-elution test, and RHD genotyping were estimated. Results: Sensitivity and specificity of the adsorption-elution test for serologically D-negative samples were 94.9% (93/98) and 91.5% (527/576), respectively. C+ phenotypes were detected in all 98 samples with DEL alleles. Despite comparable costs, RHD genotyping was more accurate and rapid than the adsorption-elution test. Conclusions: Two practical DEL typing strategies using RhCE phenotyping as an initial screening method were recommended for serologically D-negative Asian donors. Compared with DEL typing using RHD genotyping, serological DEL typing using adsorption-elution test is predicted to increase the incidence of anti-D alloimmunization and decrease the D-negative donor pool without having any cost-competitiveness but can be used in laboratories where molecular methods are not applicable.

Rodent stem cells demonstrated regenerative effects in diabetic neuropathy via improvement in nerve perfusion. As a pre-clinical step, we explored if human mobilized mononuclear cells (hMNC) would have the same effects in rats. hMNC were injected into Rt. hind-limb muscles of streptozotocin-induced diabetic nude rats, and the grafts were monitored using with MRI. After 4 weeks, the effects were compared with those in the vehicle-injected Lt. hind limbs. Nerve conduction, muscle perfusion and gene expression of sciatic nerves were assessed. Induction of diabetes decreased nerve function and expression of Mpz and Met in the sciatic nerves, which are related with myelination. hMNC injection significantly improved the amplitude of compound muscle action potentials along with muscle perfusion and sciatic nerve Mpz expression. On MRI, hypointense signals were observed for 4 weeks at the graft site, but their correlation with the presence of hMNC was detectable for only 1 week. To evaluate paracrine effects of hMNC, IMS32 cells were tested with hepatocyte growth factor (HGF), which had been reported as a myelination-related factor from stem cells. We could observe that HGF enhanced Mpz expression in the IMS32 cells. Because hMNC secreted HGF, IMS32 cells were co-cultured with hMNC, and the expression of Mpz increased along with morphologic maturation. The hMNC-induced Mpz expression was abrogated by treatment of anti-HGF. These results suggest that hMNC could improve diabetic neuropathy, possibly through enhancement of myelination as well as perfusion. According to in vitro studies, HGF was involved in the hMNC-induced myelination activity, at least in part.

Objectives This study was conducted to establish the Korean national registry, to evaluate the distribution of unexpected antibodies, and to determine the frequencies of specific antigen-negative blood units. Methods Data added to the Korean national registry between July 2013 and April 2016 were analyzed. The distribution of unexpected antibodies and frequencies of specific antigen-negative blood units were estimated. Results In total, 3,513 cases from 22 institutes were registered. The most common single alloantibodies were anti-E, anti-Lea, and anti-M. The most common multiple alloantibodies were anti-E with anti-c, anti-C with anti-e, and anti-Lea with anti-Leb. The frequencies of E-, Lea-, and M-negative units were 42.3%, 56.9%, and 20.2%, respectively. Conclusions The distribution of unexpected antibodies and frequencies of specific antigen-negative blood units were investigated using data from the Korean national registry. The results provide useful data to predict the number of blood units to be tested to obtain compatible blood units.

Background The first regulation on haemovigilance in South Korea was first established in 1999 by the Blood Management Act. The purpose of this act was mainly focused on management of transfusion-transmitted infections. Aims A more general form of haemovigilance system was needed for surveillance on the transfusion chain. Materials & Methods A research funded by the government was initiated in 2007 for development of a haemovigilance system. Review on haemovigilance systems of other countries and development of draft report forms were proceeded on the first year. Pilot operation of the system with 46 voluntary participating institutions was conducted during 2008-2009 including development of an online reporting system via the website. Results The system was formally launched in a nationwide scale in 2010, financially supported continuously by the government but independently operated by the Korean Society of Blood Transfusion. The program is a centralized, stand-alone, voluntary reporting, non-punitive, anonymized, comprehensive system. The number of reporting hospitals and reports submitted has grown every year, with a significant increase after several administrative changes regarding financial incentives and institutional accreditation programs which requested the participation of haemovigilance. Workshops are held for participants every year, and the annual reports can be accessed on the website. Among 215 participating hospitals, 169 reported 3,375 adverse events during the fiscal year 2016. Conclusion We have successfully operated the program for ten years, including a two-year pilot study. We hope to further enhance the system qualitatively regarding case validation and standardization of case reporting.

Utilization Analysis of Neonatal Red Blood Cell Aliquots and Development of Electronic Issuing System Minjeong Nam, Mi Jung Kim, Soon He Choi, Yun Ji Hong, Hyungsuk Kim, Sang Mee Hwang, Jeong Su Park, Chang Won Choi, Kyoung Un Park, Junghan Song, Kyou Sup Han Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Departments of Laboratory Medicine and Pediatrics, Seoul National University Bundang Hospital, Seongnam, Korea

It is often difficult for standard blood banks in Korea to supply adequate amounts of blood for patients with rare phenotype. Moreover, the definition of a blood in need is ambiguous, and much remains to be learned. In this study, we determined the prevalence of various red blood cell (RBC) antigens from a donor viewpoint and estimated the demand for specific antigen-negative blood from a patient viewpoint. Our data will aid the establishment of a Rare Blood Program in Korea (KRBP). RBC genotyping of 419 blood donors was performed using a Lifecodes RBC/RBC-R typing kit (Immucor, Norcross, GA). A national recipient registry website has been established. Each hospital-based blood bank voluntarily enters data on antibodies detected and identified and the outcomes of specific antigen testing. We calculated the availabilities of specific antigen-negative blood components based on these registry data and predicted the prevalence of RBC antigens via RBC genotyping. The prevalences of various RBC antigens in the D-negative population were determined for the first time, and the Cartwright, Scianna, Dombrock, Colton, Landsteiner-Wiener, Cromer, and Knops blood group systems were identified. The availabilities of specific antigen-negative units differed when calculations were based on serotyping or genotyping, especially in the D-negative group. Data on the prevalences of various blood antigens are essential for estimating the availabilities of blood components that are appropriate for use by patients expressing relevant antibodies. Then, blood banks would be able to efficiently supply safe blood products.

Disseminated intravascular coagulation (DIC) is characterized by consumption of coagulation factors and anticoagulants. Thrombin generation assay (TGA) gives useful information about global hemostatic status. We developed a new TGA system that anticoagulant addition can deplete thrombin generation in plasma, which may reflect defective anticoagulant system in DIC. TGAs were measured on the calibrated automated thrombogram with and without thrombomodulin or protein Z in 152 patients who were suspected of having DIC, yielding four parameters including lag time, endogenous thrombin potential, peak thrombin and time-to-peak in each experiment. Nonsurvivors showed significantly prolonged lag time and time-to-peak in TGA-protein Z system, which was performed with added protein Z. In multivariate Cox regression analysis, lag time and time-to-peak in TGA system were significant independent prognostic factors. In TGA-protein Z system, lag time and time-to-peak were revealed as independent prognostic factors of DIC. Protein Z addition could potentiate its anticoagulant effect in DIC with poor prognosis, suggesting the presence of defective protein Z system. The prolonged lag time and time-to-peak in both TGA and TGA-protein Z systems are expected to be used as independent prognostic factors of DIC.

As an East-Asian international study, we evaluated erythrocyte alloimmunity by gender and history of transfusion or pregnancy. In total, data from more than 1,826,000 patients were analyzed, from whom 26,170 irregular erythrocyte antibodies were detected in 22,653 cases. Antibody frequencies in these cases were as follows: anti-E, 26.8%; anti-Lea, 20.0%; anti-P1, 7.1%; anti-M, 6.4%; anti-Mia, 5.6%; anti-c + E, 5.6%; anti-Leb, 4.6%; anti-D, 2.8%; anti-Fyb, 2.6%; anti-Lea+Leb, 2.5%; anti-Dia, 2.0%; and others. For pregnant patients, anti-D (12.7%) was statistically more frequent. For transfused patients, anti-E (37.3%), anti-c + E (9.5%), anti-C + e (3.3%) and anti-Jka (3.1%) were significantly more frequent.

This study was conducted to establish the Korean national registry, to evaluate the distribution of unexpected antibodies, and to determine the frequencies of specific antigen-negative blood units.Data added to the Korean national registry between July 2013 and April 2016 were analyzed. The distribution of unexpected antibodies and frequencies of specific antigen-negative blood units were estimated.In total, 3,513 cases from 22 institutes were registered. The most common single alloantibodies were anti-E, anti-Lea, and anti-M. The most common multiple alloantibodies were anti-E with anti-c, anti-C with anti-e, and anti-Lea with anti-Leb. The frequencies of E-, Lea-, and M-negative units were 42.3%, 56.9%, and 20.2%, respectively.The distribution of unexpected antibodies and frequencies of specific antigen-negative blood units were investigated using data from the Korean national registry. The results provide useful data to predict the number of blood units to be tested to obtain compatible blood units.

Background: In the Korean population, gene frequencies of Human Neutrophil Alloantigen-1 (HNA-1), HNA-2, HNA-4, and HNA-5 were determined using serological and genotyping methods. However, no study assessing the gene frequencies of HNA-3 among the Korean population has been conducted. The aims of this study were to report the gene frequencies of HNA-3, to estimate the risk of HNA-3a (or HNA-3b) alloimmunization, and to secure donors of granulocytes for anti-HNA-3 antibodies among the Korean population. Methods: Genotyping of HNA-3a and HNA-3b genes of 110 healthy and unrelated Korean donors was performed using a polymerase chain reaction with sequence-specific primers. Results: The observed frequencies of HNA-3a and HNA-3b were 0.695 and 0.305, respectively, among the Korean population. The HNA-3a gene frequency of Koreans was significantly lower than those of American Whites and Germans (P＜0.05). The risk of HNA-3a and HNA-3b alloimmunization due to pregnancy (transfusion) was 0.065 (0.084) and 0.147 (0.250), respectively, among the Korean population. The risk of HNA-3a alloimmunization was significantly higher in the Korean population than in the German and American White populations (P＜0.05). Conclusion: The gene frequencies of HNA-3 and the risk of HNA-3a (HNA-3b) alloimmunization due to pregnancy or transfusion among the Korean population were determined. We also identified individuals who were HNA-3a/3a or HNA-3b/3b for the granulocyte panel which could be used for anti-HNA antibody identification. (Korean J Blood Transfus 2015;26:185-192)

Background and Aims Protein Z (PZ) is an anticoagulant that accelerates the inhibitory effect of PZ-dependent protease inhibitor (ZPI) on coagulation factor Xa. We assessed functional status of PZ system in 158 patients with liver cirrhosis and 59 healthy controls. Methods Plasma PZ and ZPI levels were measured by enzyme immunoassay. Thrombin generation assays (TGA) were performed with and without thrombomodulin (TM) or PZ, and the ratios were calculated by dividing TGA values with TM or PZ by values without TM or PZ. Results PZ and ZPI levels were reduced and elevated in advanced cirrhosis, respectively. The lag time ratio–PZ was significantly higher in cirrhosis patients than controls and correlated with the model for end-stage liver disease score. The peak thrombin ratio–PZ and endogenous thrombin potential (ETP) ratio–PZ were significantly lower in cirrhosis patients than controls and correlated with the severity of liver cirrhosis. The peak thrombin ratio–PZ was dramatically reduced in advanced cirrhosis. Cirrhosis patients had a significantly higher ETP ratio–TM than the controls, although the ratio was not correlated with cirrhosis severity. The lag time ratio–PZ and peak time ratio–PZ were significantly correlated with the levels of all coagulation and anticoagulation factors. Interestingly, the lag time ratio–PZ and peak thrombin ratio–PZ were significantly associated with thrombotic events. Conclusion The anticoagulant role of PZ is insufficient in advanced stages of cirrhosis. Our newly developed functional assay for measuring the PZ system is expected to reflect the ongoing hypercoagulability of cirrhosis.

Background Autologous serum eye drops (ASEDs) have been used to treat many eye diseases. However, there are no standardized guidelines for the production and quality control (QC) of ASEDs in Korea. Our aim was to propose standardized guidelines for the production and QC of ASEDs. Study Design and Methods We conducted a nationwide survey consisting of questions regarding the methods used in each hospital for the production and QC of ASEDs. The survey was sent by e-mail to 89 doctors responsible for the blood banks at different hospitals. Results Thirty-two hospitals replied, and 13 hospitals reported using the ASEDs in the treatment of patients with eye diseases. The screening test for patients, amount of blood sampling, type of bottle used for blood collection, details about the production of the eye drops, and storage methods and shelf life of unopened and opened bottles of eye drops varied between hospitals. Conclusion Based on an analysis of the survey results and a review of the standard operating procedures and protocols for ASEDs used in Japan, Germany, England and Wales, and the United States, we proposed standardized guidelines for the production and QC of ASEDs in Korea. ASEDs are not cell therapy products in the strictest sense. However, because eye drops are composed of serum isolated from blood and are used in patients, we consider ASEDs to be the basis for cell therapy products. Therefore, ASEDs should be produced and stored according to standardized guidelines based on the Good Manufacturing Practice guidelines.

We compared preoperative autologous blood donation (PABD) using serial manual whole blood (WB) and PABD using a single session, double-unit erythrocytapheresis in terms of the hemodynamic recovery and clinical outcomes.This study included 56 donors in the WB PABD group and 117 donors in the double-unit erythrocytapheresis PABD group. All subjects were men with body weight70 kg, Hb level13.3g/dL, Hct40%, and who were scheduled for oral and maxillofacial surgery. Three cycles of manual WB collection for PABD or a single session, double-unit erythrocytapheresis using the Alyx was performed.There were no significant differences in donor demographic variables including age, height, weight, Hb, Hct, or red cell mass between the 2 groups. The double-unit erythrocytapheresis was completed earlier than the last manual WB PABD (at 15.3 ± 4.7 days and 6.5 ± 3.2 days before surgery, p0.001). Hct values before surgery were higher in the double-unit erythrocytapheresis PABD group than in the manual WB PABD group (39.7 ± 3.2 vs. 38.6 ± 2.7, p=0.024). ΔHct and %ΔHct before the first PABD and before surgery were lower in the double-unit erythrocytapheresis PABD group than in the manual WB PABD group (-5.6 ± 2.8 vs. -6.8 ± 2.7, p=0.010 and -12.3 ± 5.9 vs. -14.8 ± 5.6, p=0.008, respectively). The incidence of additional allogeneic blood transfusions during or after surgery and the post-operative Hb and Hct values were similar in the 2 groups. The length of hospital stay after surgery was significantly longer in the manual WB PABD group than in the double-unit erythrocytapheresis group (6.1 ± 2.5 vs. 5.4 ± 1.9, p=0.043). Of the 33 donors in the double-unit erythrocytapheresis PABD group, 7 (21.2%) reported discomforts related to the procedure, and 6 graded the discomforts (hypocalcemia, perioral tingling sense, paresthesia, dizziness, stuffiness, pain on the intravenous site, and muscle tension) as mild.The single session, double-unit erythrocytapheresis prolonged the time interval between PABD and surgery and led to better hemodynamic recovery than the serial manual WB PABD, and hypocalcemic symptoms were mild.

Background Monocytes express substantial amounts of thrombomodulin, which is consumed throughout ongoing thrombin generation. The modulation of thrombomodulin may aggravate intravascular fibrin deposition and the clinical course of disseminated intravascular coagulation (DIC). Although thrombomodulin restoration has received considerable attention, no reports have been published on the in vivo expression status of thrombomodulin. CD14 expression on monocytes is important for regulation of the inflammatory response. We used an ex vivo stimulation study to evaluate the association of the levels of monocyte-expressed thrombomodulin and CD14 messenger RNA (mRNA) with the severity and prognosis of disseminated intravascular coagulation. Methods A total of 78 patients with suspected DIC were enrolled. Thrombomodulin and CD14 mRNA levels were measured in peripheral blood by real-time quantitative reverse-transcription polymerase chain reaction. Thrombomodulin and CD14 mRNA were also assessed in ex vivo cultures of peripheral whole blood that were stimulated by lipopolysaccharide or thrombin. Results The levels of monocyte-expressed thrombomodulin mRNA were significantly lower in the non-survivors than in the survivors. A low level of monocyte-expressed thrombomodulin mRNA was a significant prognostic marker, but CD14 did not possess prognostic power. Monocyte-expressed CD14 mRNA correlated significantly with the severity of DIC in survivors. In addition, stimulation of ex vivo cultures of whole blood demonstrated that thrombin upregulates both thrombomodulin and CD14 mRNA, and lipopolysaccharide downregulates thrombomodulin mRNA. Conclusions The downregulation of thrombomodulin on monocytes reflects the decompensated status of physiological defenses against hypercoagulopathy and represents the poor prognosis in DIC. The expression levels of thrombomodulin on monocytes may be a useful marker to screen for candidates eligible for recombinant thrombomodulin therapy in future.

Background The inventory size for cord blood (CB) depends on the ethnic diversity of human leukocyte antigen (HLA) and the size estimation is important for public health in each ethnicity. Study Design and Methods We estimate the CB inventory size in Koreans with stored CB units (CBUs) and patients who underwent allogeneic hematopoietic stem cell transplantation. Two-digit HLA specificities were determined using intermediate DNA typing. From 17,508 stored Korean CBUs, 1460 haplotypes with a frequency greater than 0.001% were used for reconstitution of the HLA. A total of 1002 transplanted patients' HLA was used for matching probability calculation. Results The best probability for 6/6 matching is 47% in 500,000 hypothetical size. Ninety-five percent probability is achieved with 51,000 CBUs in 5/6, and 2150 in 4/6 matching condition. Because 4/6 matched CB is rarely selected in the Korean situation, 51,000 units is the lowest limit of CBUs required and the number will be adjusted depending on the cell number required for patients and the resolution of HLA typing. Conclusion Approximately 51,000 units could provide the minimum requirement for hematopoietic transplantation in Korea.

S. Panzer, S. Engelbrecht, M. F. Cole-Sinclair, E. M. Wood, S. Wendel, S. Biagini, Z. Zhu, J.-J. Lefrere, G. Andreu, T. Zunino, J.-J. Cabaud, P. Rouger, O. Garraud, K. Janetzko, M. Muller-Steinhardt, P. van der Burg, A. Brand, P. Agarwal, T. Triyono, A. Gharehbaghian, N. Manny, O. Zelig, A. Takeshita, Y. Yonemura, H. Fujihara, K. E. Nollet, H. Ohto, K.-S. Han, V. S. Nadarajan, G. Berlin, S. G. Sandler, R. G. Strauss & H. W. Reesink

Antiphospholipid syndrome (APS) is an autoimmune disease in which antiphospholipid antibodies (aPLs) are generated. Previous studies show concurrence of APS and thrombotic thrombocytopenic purpura; therefore it is plausible to assume that anti-ADAMTS13 autoantibody is also involved in the pathophysiology of APS. We investigated the clinical significance of ADAMTS13 activity and anti-ADAMTS13 antibody in patients with aPLs. Two hundred and sixteen patients with positive lupus anticoagulant and/or anticardiolipin antibody were included. ADAMTS13 activity and anti-ADAMTS13 antibody were measured using fluorescence resonance energy-transfer technology and ELISA, respectively. Reduced ADAMTS13 activity was observed in 40.3% (87/216) of patients with aPLs. Although 33.8% (73/216) of patients were positive for anti-ADAMTS13 antibody, 41 of these 73 patients had normal levels of ADAMTS13 activity. Reduced ADAMTS13 activity was a significant risk factor for thrombotic events. Thrombotic events and age contributed to the reduced level of ADAMTS13 activity. Presence of anti-ADAMTS13 antibody did not show any association with the level of ADAMTS13 activity. Patients with autoimmune diseases tended to show higher levels of anti-ADAMTS13 antibody. Our findings suggest that reduced ADAMTS13 activity is a significant thrombotic risk factor in patients with aPLs irrespective of the presence of anti-ADAMTS13 antibody. Presence of anti-ADAMTS13 antibody is not seen with reduced activity and it tends to be increased in patients with autoimmune diseases.

We performed a retrospective study to predict the risk factors related to general discomfort, limitations in activities of daily living (ADLs), and intention of a second donation in hematopoietic stem cell (HSC) donation. The subjects of this study were 1,868 consecutive unrelated volunteer donors who donated HSCs through the Korea Marrow Donor Program between November 2007 and April 2014. Bone marrow (BM) collections were performed without administration of granulocyte-colony stimulating factor (G-CSF) and peripheral blood stem cell (PBSC) collections were performed after mobilization with G-CSF administered subcutaneously for 4 or 5 consecutive days at a daily dose of around 10 mcg/kg of body weight. General discomfort and limitations in ADLs were assessed by numerical measurement (scores of 0 to 10), and donor's intention of a second donation by yes or no reply. The post-donation questionnaires were completed within 48 hours after HSC collection, and at 1 week, 4 weeks, and 4 months thereafter. Pre-donation and post-donation predictors of general discomfort and limitations in ADLs were identified using univariate and multivariate linear regression considering donor demographic and clinical characteristics (age, sex, collection method, collection period) and the intention of a second donation on day 1. Pre-donation and post-donation predictors of the intention of a second donation were identified using univariate and multivariate logistic regression considering donor demographic and clinical characteristics (age, sex, collection method, collection period). Men represented 76.77% of donors. The median age at donation was 28 years (range, 19-51), and about 93.00% of donors were younger than 40 years of age. Among HSC collection, 61.88% were completed in a day. PBSC and BM collections accounted for 94.49% and 5.51% of HSC collections, respectively. Predictors of general discomfort included female sex (P< 0.0001), BM collection (P< 0.0001) or PBSC collection through a central line (P= 0.0349), 2-day collection (P= 0.0150), and negative or undetermined intention of a second donation on day 1 (P< 0.0001). Predictors of limitations in ADLs included age group of 30-39 years (P= 0.0046), female sex (P< 0.0001), BM collection (P< 0.0001) or PBSC collection through a central line (P< 0.0001), and negative or undetermined intention of a second donation on day 1 (P< 0.0001). The only predictor of positive intention of a second donation was male sex (P= 0.0007). Age, sex, collection method and period, and intention of a second donation should be considered risk factors when unrelated HSC donation is performed. As a collection method, PBSC collection through a peripheral line is recommended to alleviate physical stress and to improve ADLs. Female donors are more sensitive to HSC collection than male donors, and male sex of the donor is predictive of positive intention of a second donation. Disclosures No relevant conflicts of interest to declare.

Background DEL, a variant of RhD, is difficult to detect in routine blood bank testing owing to its extremely low levels of D antigen expression. However, DEL is capable of alloimmunizing a patient when transfused into RhDnegative individuals. Methods In this study, we developed real-time PCR and melting curve analysis for the rapid detection of the DEL phenotype. Results Of the 325 serologically RhD-negative individuals involved in the study, 56 (17.2%) had melting temperatures distinguishable from complete RHD absence as follows: 53 RHD (c.1227G>A) DEL specimens had a plateau at 54 - 56°C and a peak at 61.95°C, while 3 RHD (c.1222T>C) DEL had a melting temperature of 62.62°C. All DEL results were identical to those obtained by multiplex single-base primer extension reactions. Conclusions The rapid DEL genotyping method developed in this study will be useful for screening DEL in serologically RhD-negative donors and preventing the alloimmunization of RhD-negative individuals by DEL.

We hypothesized that polymorphisms of the vitamin D receptor (VDR) gene might affect clinical outcomes of allogeneic hematopoietic stem cell transplantation (HSCT). Three VDR gene polymorphisms (BsmI G>A, ApaI G>T, and TaqI T>C) were genotyped in 147 patients who underwent HLA-matched sibling allogeneic HSCT. Frequencies of infection, graft-vs.-host disease (GVHD), overall survival (OS), and disease-free survival (DFS) were compared according to genotypes and haplotypes. Infection and acute GVHD had trends to be less frequent in patients with ApaI TT genotype than non-TT genotypes (p = 0.061 and p = 0.059, respectively). For TaqI genotypes, there were no statistical differences in frequency of infection and acute GVHD (p = 0.84 and p = 0.30, respectively), but TC genotype was associated with longer OS and DFS than TT genotype (p = 0.022 and p = 0.038, respectively). In the ApaI-TaqI haplotype analysis, patients with TC haplotype had significantly longer OS and DFS than those without TC haplotype (p = 0.022 and p = 0.038, respectively). In multivariable analysis, TaqI genotype and ApaI-TaqI haplotype of recipients were independent prognostic factors for both OS and DFS. This study suggests that the genotype and haplotype of VDR in recipient might be associated with clinical outcome of sibling HLA-matched HSCT.

BACKGROUND: We analyzed neonatal factors that could affect hematopoietic variables of cord blood (CB) donated from Korean neonates. STUDY DESIGN AND METHODS: The numbers of total nucleated cells (TNCs), CD34+ cells, and CD34+ cells/TNCs of CB in neonates were compared according to sex, gestational age, birth weight, birth weight centile for gestational age, and ABO blood group. RESULTS: With 11,098 CB units analyzed, blood group O CB showed an increased number of TNCs, CD34+ cells, and CD34+ cells/TNCs compared with other blood groups. Although TNC counts were lower in males, no difference in the number of CD34+ cells was demonstrated because the number of CD34+ cells/TNCs was higher in males. An increase in the gestational age resulted in an increase in the number of TNCs and decreases in the number of CD34+ cells and CD34+ cells/TNCs. The numbers of TNCs, CD34+ cells, and CD34+ cells/TNCs increased according to increased birth weight centile as well as birth weight. CONCLUSION: CB with blood group O has unique hematologic variables in this large-scale analysis of Korean neonates, although the impact on the storage policies of CB banks or the clinical outcome of transplantation remains to be determined.

Therapeutic plasma exchange (TPE) has been used for the treatment of patients with thrombotic thrombocytopenic purpura-hemolytic uremic syndrome (TTP-HUS). We report the 10-year treatment results along with the risk factors analyses.Retrospective analyses were performed on patients who were treated with TPE for TTP-HUS.Fifty-two patients were included. Secondary causes were identified in 38 patients (73·1%). The others were classified as idiopathic. After a median five sessions of TPE, 26 patients (50·0%) achieved remission. Remission rate in patients with idiopathic and secondary TTP-HUS was 71·4 and 42·1%, respectively. Overall 30-day mortality rate was 34·6% and median overall survival was 5·2 months. Patients with hematopoietic stem cell transplantation-associated TTP-HUS did not respond and had poor overall survival. Males had a lower remission rate than females (P = 0·009).TPE was an effective treatment in patients with idiopathic TTP-HUS. Treatment results were various according to etiology and gender.

Background Heparin-induced thrombocytopenia (HIT) is an adverse drug reaction caused by antibodies to the heparin/platelet factor 4 (PF4) complex, resulting in thrombocytopenia and prothrombotic state. HIT diagnosis is challenging and depends on clinical presentation and laboratory tests. We investigated the usefulness of clinical scores and heparin/PF4 ELISA optical density (OD) as a diagnostic marker and thrombosis predictor in HIT. Methods We analyzed 92 patients with suspected HIT. The heparin/PF4 antibody was measured using a commercial ELISA kit (GTI, USA). For each patient, the 4 T's score and Chong's score were calculated. Results Of the 92 patients, 28 were anti-heparin/PF4-seropositive. The 4 T's score and Chong's score showed good correlation (r=0.874). The 4 T's score and OD values showed good performance for diagnosis of the definite and unlikely HIT groups; however, OD levels showed better sensitivity (93.8%) than the 4 T's score used alone (62.5%). Of the 92 patients, 26 developed thrombosis. The OD values were significantly higher in patients with thrombosis than in those without thrombosis (0.52 vs. 0.22, P0.4) had an increased risk of thrombosis (adjusted odds ratio 9.44 [3.35-26.6], P

Jr(a) is a high-frequency antigen found in all ethnic groups. However, the clinical significance of the anti-Jr(a) antibody has remained controversial. Most studies have reported mild hemolytic disease of the newborn and fetus (HDNF) in Jr(a)-positive patients. Recently, fatal cases of HDNF have also been reported. We report the first case of HDNF caused by anti-Jr(a) alloimmunization in twins in Korea. A 33-yr-old nulliparous woman with no history of transfusion or amniocentesis was admitted at the 32nd week of gestation because of vaginal bleeding caused by placenta previa. Anti-Jr(a) antibodies were detected in a routine laboratory examination. An emergency cesarean section was performed at the 34th week of gestation, and 2 premature infant twins were delivered. Laboratory examination showed positive direct antiglobulin test and Jr(a+) phenotype in the red blood cells and the presence of anti-Jr(a) antibodies in the serum in both neonates. The infants underwent phototherapy for neonatal jaundice; this was followed by conservative management. They showed no further complications and were discharged on the 19th postpartum day. Preparative management to ensure the availability of Jr(a-) blood, via autologous donation, and close fetal monitoring must be performed even in cases of first pregnancy in Jr(a-) women.

Platelet counting in patients with acute leukemia or disseminated intravascular coagulation (DIC) may have a risk for erroneous counts owing to the presence of nonplatelet particles or platelet activation. We evaluated automated platelet counting methods using the Abbott Cell-Dyn Sapphire (Abbott Diagnostics, Santa Clara, CA), Sysmex XE-2100 (Sysmex, Kobe, Japan), ADVIA 2120 (Siemens Diagnostics, Tarrytown, NY), and Beckman Coulter LH 750 (Beckman Coulter, Miami, FL) compared with the international reference method (IRM). Automated platelet counting methods were inaccurate compared with the IRM, without evidence of interfering nonplatelet particles. It is interesting that platelet activation markers were associated with DIC severity and erroneous platelet counting, suggesting that platelet activation is a potential source of inaccuracy. Furthermore, the artifactual in vitro platelet activation induced a high degree of intermethod variation in platelet counts. The inaccuracy of automated platelet counts increased the risk for misdiagnosis of DIC. More attention needs to be given to the accuracy of platelet counts, especially in clinical conditions with florid platelet activation.

Local anesthetics have been reported to have anticoagulant properties, but the mechanisms responsible for this action are poorly understood. Here, we evaluated the in vitro effects of 3 local anesthetics—lidocaine, ropivacaine, and bupivacaine—on the tissue factor expression by monocytes. Monocytes from peripheral blood were stimulated with lipopolysaccharide (LPS) in the presence or absence of local anesthetics. All 3 local anesthetics inhibited the expression of tissue factor antigen and tissue factor activity in LPS-stimulated monocytes in a dose- and time-dependent manner and reduced tissue factor messenger RNA (mRNA) expression in endothelial cells and a monocytic cell line. None of the 3 drugs induced apoptosis or affected the viability of monocytes. Our findings that local anesthetics inhibited the tissue factor induction in activated monocytes by inhibiting tissue factor mRNA level may demonstrate the feasibility of using local anesthetics in hypercoagulable and inflammatory conditions.

Uterine sarcomas are rare among all uterine malignancies, and frequently misdiagnosed as benign uterine diseases such as leiomyoma and adenomyosis because of lack of feasible tools for the preoperative diagnosis. Although some studies have suggested the role of serum CA-125 levels for the preoperative diagnosis, the efficacy is controversial. Since malignancy is known to be associated with systemic inflammation which leads to hematological alteration, we compared the efficacy for the preoperative diagnosis of uterine sarcomas between the neutrophil to lymphocyte ratio (NLR) and serum CA-125 levels using a case-match comparison.From November 2004 to December 2008, 55 patients with carcinosarcoma (n=21), leiomyosarcoma (n=20) and endometrial stromal sarcoma (n=14) were matched to 330 patients with leiomyoma (n=165) and adenomyosis (n=165) in terms of age at diagnosis, body mass index and uterine volume.The receiver operating characteristic curve showed the best cut-off values of the NLR (or=2.12) and serum CA-125 levels (or=27.5U/ml) for the preoperative diagnosis of uterine sarcomas, demonstrating that the NLR was more powerful for the preoperative diagnosis of uterine sarcomas than serum CA-125 levels (sensitivity, 74.5% vs. 52.3%; specificity, 70.3% vs. 50.5%; positive predictive value, 29.5% vs. 15.1%; negative predictive value, 94.3% vs. 86.5%; accuracy, 60.6% vs. 49.6%; p0.05). Furthermore, the NLR reflected recurrence and progression more accurately than serum CA-125 levels in patients with uterine sarcomas.These findings suggest that the NLR may be more useful than serum CA-125 levels as a cost-effective tool for the preoperative diagnosis in patients with uterine sarcomas.

The Lewis histo-blood group system consists of 2 major antigens-Lea and Leb-and a sialyl Lewis antigen-carbohydrate antigen (CA) 19-9. We investigated the distribution of Lewis genotypes and evaluated the relationship between the Lewis/Secretor genotypes and the serum level of CA 19-9 in a Korean population to identify whether the serum CA 19-9 levels are influenced by the Lewis/Secretor genotypes.The study included 242 individuals who had no malignancies. Lewis genotyping was performed for the 59TG, 508GA and 1067TA polymorphic sites. The Secretor genotype was determined through analysis of the 357CT and 385AT polymorphic sites and the fusion gene. Serum CA 19-9 level was analyzed using an electrochemiluminescence immunoassay.Individuals carrying the 3 common genotypes-Le/Le, Le/le(59,508), and Le/le(59,1067)-accounted for 95% of the study population. In the Korean population, the allelic frequencies of Le, Le(59), le(59,508), and le(59,1067) were 0.731, 0.010, 0.223, and 0.035, respectively. We found a significant difference in serum CA 19-9 concentrations among the 9 Lewis/Secretor genotype groups (P0.001). The serum CA 19-9 levels in subjects with genotype groups 1 and 2 (Le/- and se/se) were higher than those with genotype groups 3-6 (Le/- and Se/-; 15.63 vs 6.64 kU/L, P0.001).Le/Le, Le/le(59,508), and Le/le(59,1067) are frequent Lewis genotypes in Koreans. Because serum CA 19-9 levels are significantly influenced by the Lewis/Secretor genotypes, caution is suggested when interpreting the serum CA 19-9 levels.

Although platelet count is a good parameter for the diagnosis of disseminated intravascular coagulation (DIC), a single measurement of platelet is not enough to reflect the ongoing platelet consumption because of compensatory synthesis of circulating platelet number. Increased thrombopoiesis owing to peripheral destruction is expected in patients with DIC. Reticulated platelet, measured as immature platelet fraction (IPF), and plasma thrombopoietin (TPO) are markers of platelet production. We investigated the potential usefulness of circulating IPF and TPO in 222 patients suspected of having DIC. Both IPF and TPO levels were significantly increased in overt DIC patients and well correlated with DIC score. IPF also correlated with fibrin-related marker such as fibrinogen degradation product and D-dimer. Both IPF and TPO showed better mortality prediction than platelet count with the multivariate logistic regression and Kaplan-Meier survival analysis. These results suggest that IPF and TPO are new potential candidates to detect the severity of DIC and to predict DIC mortality.

The human umbilical cord (hUC) is a source of adult tissue-derived mesenchymal stem cells (MSCs). A pulsed low-intensity ultrasound (PLIUS) method is described for increasing the yield of MSCs from whole hUC without enzymatic digestion or growth factor supplementation. Analysis of the immunophenotype of cells and a differentiation study were performed to show the compatibility of MSCs. The mean number of cells recovered from primocultures of hUC was 6 x 10(5) cells/cm. PLIUS resulted in a 3.3-fold increase in MSC yield at passage 0. PLIUS exposure increases the yield of hUC-MSCs by promoting release and enhancing proliferation.

Thrombomodulin has a central role in the regulation of coagulation through its ability to promote generation of the potent anticoagulant, activated protein C. Aurintricarboxylic acid (ATA) has been reported to inhibit platelet function by blocking von Willebrand factor binding to platelet glycoprotein Ib and to impede thrombosis development in vivo. In the present study, we demonstrated a novel antithrombotic effect of ATA. The surface thrombomodulin expression of endothelial cells and peripheral blood monocytes was upregulated by ATA in a dose-dependent and time-dependent manner. ATA also increased the mRNA level of endothelial thrombomodulin in a dose-dependent manner. Tumor necrosis factor (TNF)-alpha (50 ng/ml) or lipopolysaccharide (20 microg/ml) downregulated the expression of endothelial thrombomodulin. Blocking of nuclear factor-kappaB by parthenolide effectively inhibited the TNF-alpha-induced thrombomodulin downregulation of endothelial cells. ATA increased endothelial thrombomodulin expression that was downregulated by TNF-alpha or lipopolysaccharide, in a dose-dependent manner. The inhibition of small G proteins of the Rho family by the Clostridium difficile toxin B-1,0643 did not increase thrombomodulin expression of endothelial cells, and ATA did not activate Rac1 in endothelial cells. These findings provide, at least in part, a novel platelet-independent mechanism of ATA that may explain the demonstrated antithrombotic efficacy of ATA.

Ael is a rare blood type which has the least amount of A antigen among A subgroups. It can be detected by special tests performed to resolve the discrepancy between red cell and serum typing in routine serological typing. The presence of A antigen on Ael red cell is demonstrable only by adsorption and elution tests. An Ael individual does not secret A substance in the saliva and may have anti-A antibody in the serum which is usually less reactive with the reagent red cells than anti-B antibody. In Korea, Ael02 has been reported more frequently than other Ael alleles. We report a case of Ael02/O04 who presented as typical phenotype O with strong anti-A and anti-B antibodies and no A antigen detected even by adsorption and elution tests. The case has been proved to be Ael02/O04 by direct sequencing analysis. In individuals with history of discrepancies in the results of ABO phenotyping, ABO genotyping is needed for an accurate evaluation of their blood type.

The CXC chemokine receptor CXCR4 and its unique ligand SDF-1 (stromal-derived factor-1) play critical roles for the retention of hematopoietic cells within the bone marrow (BM) and for their mobilization into the circulation. Lithium often produces neutrophilia in psychiatric patients, but the mechanism of mobilization related to neutrophilia has not been fully clarified. We showed here that lithium dose-dependently reduces the levels of surface CXCR4 protein and mRNA in neutrophils, but not in lymphocytes. The chemotactic migration of neutrophils in response to SDF-1 was reduced after a pre-incubation with lithium. We provide evidence that lithium down-regulates the CXCR4 expression of neutrophils and it attenuates their responsiveness to SDF-1. Our studies support the concept that down-regulation of CXCR4 is one of the mechanisms by which causes neutrophilia.

The chemokine receptor CXCR4 and its unique ligand, stromal-derived factor 1 (SDF-1), play critical roles in the retention of hematopoietic cells within bone marrow and in their mobilization into the circulation. Surface CXCR4 down-regulation in hematopoietic cells is associated with a loss of retention of the cells in bone marrow. Lipopolysaccharide (LPS), commonly referred to as endotoxin, induces neutrophilia in vivo, but the mechanism of mobilization related to neutrophilia has not been fully clarified. We show that LPS reduces CXCR4 surface expression in a dose- and time-dependent manner in neutrophils and monocytes, but not in lymphocytes. Polymyxin B neutralization of LPS in culture supernatants still induced this down-modulation, and LPS-stimulated neutrophils released interferon gamma and interleukin 1beta. These results provide evidence that CXCR4 down-regulation can be attributed to soluble factors released by neutrophils upon LPS treatment. Moreover, LPS treatment increased CXCR4 messenger RNA in neutrophils, suggesting that the down-regulation of surface CXCR4 is caused by a posttranslational mechanism, and the chemotactic migration of neutrophils in response to SDF-1 was reduced by LPS pretreatment. Thus, the present study has shown that by down-regulating neutrophil CXCR4 expression and attenuating neutrophil responsiveness to SDF-1, LPS can mobilize neutrophils from bone marrow to the peripheral blood through reducing neutrophil retention in bone marrow.

A contiguous segment attached to the cord blood unit (CBU) is required for verifying HLA types, cell viability, and, possibly, potency before transplantation since such a segment is considered to be representative of the CBU. However, little is known regarding the characteristics of contiguous segments in comparison to main bag units due to the difficulty experienced in accessing a large number of cryopreserved CBUs. In this study, we used 245 nonconforming CBUs for allogeneic transplantation. After thawing the cryopreserved CBU, the number of total nucleated cells (TNCs), CD34+ cells, and CFUs in CB from main bags and segments, as well as cell viability and apoptosis, were examined. The comparative analysis showed that the number of TNCs was significantly higher in CB from main bags, whereas the numbers of CD34+ cells and CFU-GM were significantly higher in CB from segments. While the cell viability of TNCs in segments was higher, the proportion of apoptotic TNCs was also higher. In contrast, no difference was observed between the proportion of apoptotic CD34+ cells in main bags and segments. In the correlation analysis, the numbers of TNCs, CD34+ cells, and CFU-GM in main bags were highly correlated with those in segments, indicating that CB from segments is indeed representative of CB in main bags. Taken together, we conclude that segments have higher CD34+ cells and CFU-GM and lower TNCs than the main cryopreserved bag, although the two compartments are highly correlated with each other.

BACKGROUND ABO genotyping is a useful tool in case of ABO discrepancies and in legal medicine. Recent knowledge of various alleles in the ABO gene has led to the need of a different method that can cover numerous polymorphisms. We performed a polymerase chain reaction using sequencespecific priming (PCR-SSP) with 12 primer sets and evaluated its value in the detection of 8 ABO alleles. METHODS Genomic DNA was isolated from peripheral blood of 222 unrelated Koreans. Sequencespecific primer sets for the nucleotides 261, 297, 467, 802, 803, and 1059 were selected, and 12 PCR reactions were performed for each sample. Direct sequencing was performed to evaluate the accuracy of discrimination between A(1)(Pro) and A(1)(Leu) and between O(1) and O(1v). RESULTS All the ABO genotype patterns were in an exact match with the ABO phenotypes. The results from sequencing and PCR-SSP were equivalent. The allele frequencies of A(1), B, O(1), and O(1v) were 27.25%, 19.82%, 27.25%, and 25.68% respectively. Out of total 121 A(1) alleles, 6 (4.96%) were A(1)(Pro) alleles and 115 (95.04%) were A(1)(Leu) alleles. No A(2), O(2), or CisAB alleles were found in this study. CONCLUSIONS The proportion of O(1v) allele was similar to that of O(1) allele. This was an unexpected result. We developed a method for detecting 8 ABO alleles by PCR-SSP; the method was accurate and was able to discriminate between A(1)(Pro) and A(1)(Leu) and between O(1) and O(1v).