A Study of the Molecular Characteristics of the Serologic D-- Blood Type in Korean Blood Donors / 대한수혈학회지

Background@#The Korean Red Cross has conducted serologic tests for C, c, E, e antigens and found 18 D-- donors.In this study, we performed RHCE genotyping to identify the molecular characteristics of the serologic D-- blood type in Korean blood donors. @*Methods@#We performed RHCE-specific PCR-based electrophoresis to check the amplification pattern of each exon.Sanger sequencing was conducted to find the variants in the nucleotide sequence. We determined the RHCE genotype based on the electrophoresis and Sanger sequencing results. @*Results@#Total eight out of 18 D-- donors were participated in this research. In the PCR-based electrophoresis tests, RHCE exons 3, 4, and 6 were not amplified in samples #4, #6, and #8. Also, sample #2 showed an abnormal band pattern of RHCE exon 9. The Sanger sequencing results showed that the nucleotide sequences of the RHCE exons 5, 7, and 8 in samples #4, #6 and #8 corresponded to the nucleotide sequences of RHD exons 5, 7, and 8, respectively, suggesting the possibility of a RHCE-RHD(3-8)-RHCE hybrid allele. The nucleotide sequences of RHCE exons 7 and 8 in sample #2 were the same as the nucleotide sequences of RHD exons 7 and 8, respectively.In samples #1, #3, #5, and #7, no specific variants known to cause D-- phenotype were found. @*Conclusion@#RHCE genes partially replaced by the RHD genes were found in four out of eight participants and three of them were identified as ?RHCE*02N.07, which is known as the RHCE null allele. A further study with complete RHCE sequencing could be helpful for an understanding of the molecular mechanisms of samples in which no significant variants were identified.

Current Status of Korean Red Cross HLA-Matched Platelet Donor Registry / 대한수혈학회지

HLA-matched platelet transfusion is required for patients with platelet refractoriness due to HLA alloimmunity. From 2013 to 2019, the Korean Red Cross has recruited 4,080 donors for HLA-matched platelets. The patient’s HLA information should be submitted to the Korean Red Cross in accordance with the WHO HLA serologic specificities. When HLA-matched platelets are requested, the Korean Red Cross selects the appropriate donors based on Duquesnoy’s matching grade classification (1977) and CREGs defined by Takemoto, Fuller, and Rodey (2007) and then contacts them to request blood donations. Platelets of HLA-matched donors are collected by apheresis and supplied to the hospital. To make this process more efficient, the Korean Red Cross introduced a systemic standard work procedure using a computer program for blood donor management and HLA matching. Owing to the extensive polymorphism of the HLA types, expansion of the donor pool would be required to supply HLA-matched platelets sufficiently. As the number of registered donors for HLA-matched platelets is limited, it should only be ordered when the indication criteria for its use are met. The Korean Red Cross is planning to study genotype-based matching strategies for patients with rare HLA types and receive patients’ laboratory test results from medical institutions to evaluate the effectiveness of HLA-matched platelet transfusions.

Current Status of Korean Red Cross HLA-Matched Platelet Donor Registry / 대한수혈학회지

HLA-matched platelet transfusion is required for patients with platelet refractoriness due to HLA alloimmunity. From 2013 to 2019, the Korean Red Cross has recruited 4,080 donors for HLA-matched platelets. The patient’s HLA information should be submitted to the Korean Red Cross in accordance with the WHO HLA serologic specificities. When HLA-matched platelets are requested, the Korean Red Cross selects the appropriate donors based on Duquesnoy’s matching grade classification (1977) and CREGs defined by Takemoto, Fuller, and Rodey (2007) and then contacts them to request blood donations. Platelets of HLA-matched donors are collected by apheresis and supplied to the hospital. To make this process more efficient, the Korean Red Cross introduced a systemic standard work procedure using a computer program for blood donor management and HLA matching. Owing to the extensive polymorphism of the HLA types, expansion of the donor pool would be required to supply HLA-matched platelets sufficiently. As the number of registered donors for HLA-matched platelets is limited, it should only be ordered when the indication criteria for its use are met. The Korean Red Cross is planning to study genotype-based matching strategies for patients with rare HLA types and receive patients’ laboratory test results from medical institutions to evaluate the effectiveness of HLA-matched platelet transfusions.

Evaluation of the sodium intake reduction plan for a local government and evidence-based reestablishment of objectives: Case of the Seoul Metropolitan Government / 한국영양학회지

PURPOSE: To identify the effectiveness of policy evaluation, consistent monitoring is necessary. This study aimed to carry out mid-term evaluation of objectives and programs related to comprehensive plans for sodium intake reduction by 2020 for Seoul city and then reestablish the objectives of the sodium intake reduction plans. METHODS: Literature reviews, data analysis, and reviews of expert focus-groups were performed to evaluate objectives, to develop a new goal, and to identify the priority subjects of the sodium intake reduction programs. In order to examine target populations for the programs, awareness and behaviors related to sodium intakes among Seoul citizens were examined by sex, age, and income level using the 2008~2013 Korea National Health and Nutrition Examination Survey data. RESULTS: Current objectives of the sodium intake reduction plan by 2020 for Seoul city were not appropriate, so objectives were reset to 3,600 mg of sodium intake by 2020 among Seoul citizens with 2% reduction per year. Although sodium intake showed a decreasing trend by year, it was still high, especially in men. The sodium intake reduction programs currently in progress have not been assessed at multiple levels across multiple sectors and have only been assessed fragmentarily. For dietary behavior related to sodium intakes by sex, age, and income level, sodium intake was higher in the group with less than 100 g of fruit intake compared to the group with 100 g or more. Subjects aged 30~59 years and the low household income group showed relatively higher sodium intakes. Based on the data analysis and the expert review, the priority subject of the sodium intake reduction programs was determined to be adult men. In terms of a program strategy for sodium intake reduction, multi-level and setting approaches, including work sites, home, and restaurants, were suggested to reduce sodium intakes of the target subject. CONCLUSION: The suggested objectives should be consistently monitored by data analysis, and the determined programs need to be phased in over 5 years.

Korean Red Cross HLA-Matched Platelet Donor Registry / 대한수혈학회지

BACKGROUND: Transfusion of HLA-matched platelets is required when development of platelet refractoriness occurs after repeated platelet transfusion. This study was conducted to establish a HLA-matched platelet donor registry to supply matched platelets to patients who develop platelet refractoriness. METHODS: HLA-matched platelet donors were recruited among plateletpheresis donors. HLA-A and HLA-B antigen types of recruited donors were tested using a polymerase chain reaction-sequence specific oligonucleotide probe method. RESULTS: A total of 1,029 plateletpheresis donors were recruited. HLA-A and HLA-B antigen frequencies of recruited donors were similar to those of previously reported HLA antigen frequencies of Koreans. During the study period, a patient with platelet refractoriness recovered after receiving six units of HLA-matched platelets. CONCLUSION: During this study 1,029 donors were registered as HLA-matched platelet donors and a patient with platelet refractoriness received HLA-matched platelets using this registry. Supply of HLA-matched platelets will be facilitated by continuous expansion of the number of registered HLA-matched platelet donors, development of a program for management and searching for HLA-matched donors, and establishment of a request-supply system between hospitals and the Korean Red Cross through further studies.

Development of dietary behavior items available in Korea National Health and Nutrition Examination Survey / 한국영양학회지

PURPOSE: Given that the indicators related to chronic diseases are important to evaluating goals of the national health policy, the aim of this study was to develop dietary behavior items available in the Korea National Health and Nutrition Examination Survey. METHODS: The dietary behavior items were developed based on the literature reviews, need assessment of the field, expert focus-group interviews, and expert advisory meeting. Questions for each dietary behavior item were developed by reflecting on environmental, personal, and behavior factors of the ecological frame and then revised through expert focus-group interview and expert advisory meeting. The understanding and reliability of the developed questionnaire were assessed by cognitive interview and test-retest reliability. RESULTS: The developed items were sodium and salt intake, added and simple sugar intake, enough time to consume a meal for all ages, caffeine drinks and fresh/healthy food intake for children and adolescents, and limited dietary intake, fresh/healthy food intake for seniors. In most questions except some questions on sodium and salt intake, subjects understood over 70% and consistency of responses based on the kappa values was acceptable. CONCLUSION: Developed dietary behavior items are expected to be useful for evidence-based nutrition policy, interventions and research targeting dietary patterns through investigating and monitoring dietary behavior patterns.

Evaluation of RF300 for Leukoreduction of Red Blood Cells / 대한수혈학회지

BACKGROUND: Use of universal leukoreduction for prevention of leukocyte associated transfusion reactions is common practice in many countries. This study was conducted in order to evaluate the performance of a newly developed leukoreduction filter for red blood cells (RBCs), the RF300 (Kolon Industries, Inc, Gumi, Korea). METHODS: Filtration time, RBC recovery, residual leukocyte count, and leukocyte removal rate were evaluated. To assess the quality of RBCs after filtration, percent hemolysis was monitored for a period of 21 days. Performance of the RF300 (N=78) was compared with that of the Bio-R O2 plus (Fresenius, Hamburg, Germany), the Pall Purecell RC (Pall Co., Washington, USA), and the Sepacell R-500N (Asahi, Tokyo, Japan). RESULTS: The shortest filtration time was observed using the RF300 (P<0.05). Using the RF300, recovery of RBC was 96.5%, which was higher than that of two filters (P<0.05). Mean residual leukocyte count was 0.26x10(6)/unit, with a leukocyte removal rate of 3 log. Using the RF300, mean percent hemolysis was 0.32% at day 21, which was comparable with that of two filters, but lower than that of one filter (P<0.05). CONCLUSION: The RF300 meets all established quality requirements for conduct of safe and effective leukoreduction of RBCs.

Complete Sequencing of the ABO Alleles of the Common ABO Group and Rare ABO Subgroups in Koreans / 대한수혈학회지

BACKGROUND: Complete sequencing, except for intron 1, of the ABO allele in some populations has been reported. However, so far, one report on complete sequencing of the ABO gene in three Korean families with normal ABO phenotypes has been published. This study aimed to establish a reference database of common ABO alleles in Koreans. METHODS: Screening of common ABO alleles, including homozygote form, was performed by direct sequencing of exons 6 and 7 and by real-time PCR using displacing probes in 95 healthy donors. Genomic DNA from the common ABO group (n=8) and some ABO subgroups (n=7) was used in complete sequencing (except for intron 1) of the ABO allele. RESULTS: The sequences of B101/B101 (n=1), O01/O01 (n=1) were identical with the corresponding sequences registered in Genebank. A102 and A105 had a common point mutation, 1142 C>T in intron 4. A102 (n=3/11) and O02 (n=3/3), selected by sequencing of exons 6 and 7, were reclassified into A105 and O65 by whole genomic sequencing, respectively. Analytic results for ABO subgroups were as follows: B3, B101/O01 (n=3) and B101/O02 (n=1); A1B3, A102/B101 (n=1) and A105/B101 (n=1); Ax, A102/O01 (n=1). CONCLUSION: We established a reference database of common ABO alleles in Koreans and found that the molecular basis of introns of ABO alleles in the Korean population differs from that reported in previous studies of other populations.

Comparison of Methods for Detecting Bacterial Contamination in Platelet Concentrates / 대한수혈학회지

BACKGROUND: Bacterial contamination of platelets represents the highest infectious risk for a transfusion. In this study, we evaluated 2 culture-based systems that have been approved by the US FDA for bacterial screening. METHODS: Platelet concentrates were inoculated with 5 bacterial species to give a final concentration of 10(0), 10(1) and 10(2) CFU/mL. Samples for culture were taken immediately after inoculation (0 hr sample) and after 24 hrs (24 hr sample). For the BacT/ALERT 3D system, a 10 mL sample was inoculated into an aerobic culture bottle and incubated for 7 days. For the Pall eBDS system, 3 mL samples were taken from the 0 hr and 24 hr samples, respectively. The samples were incubated for 24 hrs and 30 hrs. RESULTS: Both systems detected all inoculated units both in the 0 hr and 24 hr samples, except for units inoculated with K. pneumoniae. Eleven units out of 30 units inoculated with K. pneumoniae were detected by the BacT/ALERT 3D system in the 24 hr samples. The Pall eBDS system detected 8 of 30 units in the 24 hr samples. CONCLUSION: Implementation of either system will decrease the risk of transfusing bacterially contaminated platelets. However, testing for bacterial contamination will not completely prevent septic transfusion reactions; pathogen inactivation that is now available should also be considered as an alternative method to reduce the risk of bacterial contamination.

Evaluation of Single-use Blood Donor Arm Antiseptic Kits / 대한수혈학회지

BACKGROUND: Transfusion-associated bacterial contamination (TABC) have become one of the major hazards of transfusion and the donors' normal skin flora is considered to be the most important source of contamination. We therefore evaluated the effectiveness of single-use blood donor arm antiseptic kits available on the Korean market. METHODS: Forty-eight antiseptic kits of two manufacturers consisting of 70% isopropyl alcohol swab stick and 10% povidone iodine swab stick were evaluated for their performance in reducing the bacterial load of the skin before and after disinfection. Staff response about ease of use was also evaluated. RESULTS: Whereas before disinfection the mean bacterial count was 5 CFU/plate or less, after disinfection bacterial growth was hardly observed. Overall both antiseptic kits showed a percent reduction of more than 98% compared to predisinfection. The amount of antiseptic solution of one manufacturers' kit was excessive and leakage of the antiseptic solution was also observed. CONCLUSION: The effectiveness of both antiseptic kits evaluated in this study was satisfactory to be used for blood donors, but an improvement to the packaging of one manufacturers' kit is required before it can be used in the field.

Cryopreservation and Thawing of Red Blood Cells Using Haemonetics ACP 215 / 대한진단검사의학회지

BACKGROUND: The FDA has approved the storage of frozen red blood cells (RBCs) at -80degrees C for 10 years. After deglycerolization, the RBCs can be stored at 4degrees C for no more than 24 hours, because open systems are currently being used. We evaluated Haemonetics ACP 215, an automated, functionally closed system, for both the glycerolization and deglycerolization processes. METHODS: Thirty packed RBCs that had been glycerolized and stored at -80degrees C for 2 weeks were thawed, deglycerolized and resuspended in AS-3. The RBCs were then stored at 4degrees C for 2 weeks. For the evaluation of the procedure, RBC recovery rate, osmolarity, specific gravity, LDH, K+, Hb-2, 3 DPG, Hb-ATP, and plasma hemoglobin were tested at day 0 and day 14. RESULTS: The recovery rate of RBCs was 83.7+/-2.6% (78.9-88.8%). The Hb ATP and 2, 3-DPG of RBCs were 5.16+/-1.0 mol/g Hb and 10.4+/-2.4 mol/g Hb, respectively, at day 0. The supernatant K+, specific gravity, osmolarity, LDH were 1.3+/-0.6 mmol/L, 1.008+/-0.001, 295.0+/-3.1 mOsm/kgH2O, 175.0+/-39.0 unit/L, respectively. All measurements were acceptable to allow the RBCs deglycerolized on ACP 215 to be stored at 4degrees C for 14 days. The blood cultures were negative at day 0 and day 14. CONCLUSIONS: Haemonetics ACP 215 provides a closed, automated system for RBC glycerolization and deglycerolization. This study showed that the RBCs that were glycerolized and deglycerolized in the automated instrument and stored in AS-3 at 4degrees C for 14 days are of an acceptable quality.

ABO Gene Analysis of Discrepant ABO Blood Group in Blood Donors / 대한수혈학회지

BACKGROUND: An exact ABO blood group is essential for prevention of transfusion accident and safe transfusion therapy. It is known that one of causes of ABO discrepancies is ABO subgroup caused by genetic polymorphism. Therefore, we analyzed ABO genotype of ABO discrepancies in blood donors and studied the distribution and cause of ABO discrepancies. METHODS: This study examined 118 samples showing ABO discrepancies of ABO blood typing between May 2003 and Dec 2003. ABO genotyping using the polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) method was performed on 118 samples. Restriction enzymes including BssH II, Kpn I and Alu I were used for PCR-RFLP. RESULTS: The genotypes of 118 cases were composed of 43 cases of A/B, 12 cases of A/O, 10 cases of B/O, 1 case of B/B, 37 cases of cis-AB/O, 4 cases of cis-AB/A, 11 cases of cis-AB/B. The genotype of cis-AB/O showed 32 cases with phenotype A2 B3 , 2 cases with phenotype A2 B, 2 cases with phenotype A1 B3 , 1 case with phenotype Ael B. The genotype of cis-AB/B showed 11 cases with phenotype A2 B, and cis-AB/A showed 2 cases with phenotype A2 B3 , 1 case with phenotype A1 Bx and 1 case with phenotype A1 Bel. CONCLUSION: These data demonstrated that the most frequent genotype of ABO discrepancies in our study is cis-AB. The most predominent phenotype of cis-AB/O is A2 B3 . ABO genotyping is useful in resolving ABO discrepancies, and determination of ABO subgroups.