Korean clinical practice guidelines for the diagnosis of hereditary hemolytic anemia

Although the prevalence of hereditary hemolytic anemia (HHA) is relatively low in Korea, it has been gradually increasing in recent decades due to increment in the proportions of hemoglobinopathies from immigrants of South East Asia, raising awareness of the disease among clinicians, and advances in diagnostic technology. As such, the red blood cell (RBC) Disorder Working Party (WP), previously called HHA WP, of the Korean Society of Hematology (KSH) developed the Korean Standard Operating Procedures (SOPs) for the diagnosis of HHA in 2007. These SOPs have been continuously revised and updated following advances in diagnostic technology [e.g., flow cytometric osmotic fragility test (FOFT) and eosin-5-maleimide (EMA) binding test], current methods for membrane protein or enzyme analysis [e.g., liquid chromatography-tandem mass spectrometry (LC-MS/MS), ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), high-performance liquid chromatography (HPLC)], and molecular genetic tests using next-generation sequencing (NGS). However, the diagnosis and treatment of HHA remain challenging as they require considerable experience and understanding of the disease. Therefore, in this new Korean Clinical Practice Guidelines for the Diagnosis of HHA, on behalf of the RBC Disorder WP of KSH, updated guidelines to approach patients suspected of HHA are summarized. NGS is proposed to perform prior to membrane protein or enzyme analysis by LC-MS/MS, UPLC-MS/MS or HPLC techniques due to the availability of gene testing in more laboratories in Korea. We hope that this guideline will be helpful for clinicians in making diagnostic decisions for patients with HHA in Korea.

Establishment of Pediatric Reference Intervals for Routine Laboratory Tests in Korean Population: A Retrospective Multicenter Analysis

Background@#Reference intervals defined for adults or children of other ethnicities cannot be applied in the evaluation of Korean pediatric patients. Pediatric reference intervals are difficult to establish because children are in their growing stage and their physiology changes continuously. We aimed to establish reference intervals for routine laboratory tests for Korean pediatric patients through retrospective multicenter data analysis. @*Methods@#Preoperative laboratory test results from 1,031 pediatric patients aged 0 month–18 years who underwent minor surgeries in four university hospitals were collected. Age- and sex-specific reference intervals for routine laboratory tests were defined based on the Clinical and Laboratory Standards Institute (CLSI) EP28-A3c guidelines. @*Results@#The pediatric reference intervals determined in this study were different from existing adult reference intervals and pediatric reference intervals for other ethnicities. Most tests required age-specific partitioning, and some of those required sex-specific partitioning for at least one age-partitioned subgroup. Erythrocyte sedimentation rate, monocyte percentage, basophil percentage, activated partial thromboplastin time, glucose, cholesterol, albumin, bilirubin, chloride, and C-reactive protein did not show any difference between age- or sex-partitioned subgroups. @*Conclusions@#We determined Korean pediatric reference intervals for hematology, coagulation, and chemistry tests by indirect sampling based on medical record data from multiple institutions. These reference intervals would be valuable for clinical evaluations in the Korean pediatric population.

Mutation of ten-eleven translocation-2 is associated with increased risk of autoimmune disease in patients with myelodysplastic syndrome

Background/Aims@#Myelodysplastic syndrome (MDS) is caused by genetic and epigenetic alteration of hematopoietic precursors and immune dysregulation. Approximately 20% of patients with MDS develop an autoimmune disease (AID). Here, we investigated whether particular genetic mutations are associated with AID in patients with MDS. @*Methods@#Eighty-eight genetic mutations associated with myeloid malignancy were sequenced in 73 MDS patients. The association between these mutations and AID was then analyzed. @*Results@#The median age of the 73 MDS patients was 70 years (interquartile range, 56 to 75), and 49 (67.1%) were male. AID was observed in 16 of 73 patients (21.9%). Mutations were detected in 57 (78.1%) patients. The percentage (68.8% vs. 80.7%, p = 0.32) and the mean number of mutations (1.8 ± 1.6 vs. 2.2 ± 1.8, p = 0.34) in MDS patients with or without AID were similar. However, the ten-eleven translocation- 2 (TET2) mutation rate was significantly higher in patients with AID than in those without (31.3% vs. 5.3%, respectively; p = 0.001). All TET2 mutations were variants of strong clinical significance. @*Conclusions@#Mutation of TET2 in patients with MDS may be associated with increased risk of developing AID.

Classification of acute myeloid leukemia

The World Health Organization (WHO) Classification of Tumors of Hematopoietic and Lymphoid Tissues was revised in 2017 on the basis of recent high-throughput sequencing and gene expression data on hematologic malignancies. This review explores the current WHO classification of acute myeloid leukemia (AML) and related precursor neoplasms, highlighting the changes made in the current edition and focusing on the diagnosis of AML.

A Case of Fungemia with Co-isolation of Candida parapsilosis and Trichosporon asahii Confirmed by MALDI-TOF MS and D2 rRNA Sequencing

Fungi are a major cause of human infections with diverse clinical manifestations. The incidence of fungal infections has increased over time, particularly in patients who have risk factors such as neutropenia, immune suppression, an intravascular catheter, parenteral nutrition, a prosthetic device, and prior broad spectrum antibiotic therapy. Here, we present an unusual case of co-infection by 2 distinct fungi, Candida parapsilosis and Trichosporon asahii, isolated from a patient who did not have any known risk factors initially, except active pulmonary tuberculosis. Despite the negative conversion of sputum acid-fast bacilli (AFB) culture test after treatment, clinical symptoms were refractory to therapy. The patient developed symptoms suggesting septic shock, and 2 distinct colonies were isolated from a blood specimen, which were identified as C. parapsilosis and T. asahii by MALDI-TOF and rRNA sequencing. Fever and hypotension were relieved after anti-fungal agent injection, and pulmonary lesions identified by imaging also improved.

Comparison of Ion Personal Genome Machine Platforms for the Detection of Variants in BRCA1 and BRCA2 / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: Transition to next generation sequencing (NGS) for BRCA1/BRCA2 analysis in clinical laboratories is ongoing but different platforms and/or data analysis pipelines give different results resulting in difficulties in implementation. We have evaluated the Ion Personal Genome Machine (PGM) Platforms (Ion PGM, Ion PGM Dx, Thermo Fisher Scientific) for the analysis of BRCA1/2. MATERIALS AND METHODS: The results of Ion PGM with OTG-snpcaller, a pipeline based on Torrent mapping alignment program and Genome Analysis Toolkit, from 75 clinical samples and 14 reference DNA samples were compared with Sanger sequencing for BRCA1/BRCA2. Ten clinical samples and 14 reference DNA samples were additionally sequenced by Ion PGM Dx with Torrent Suite. RESULTS: Fifty types of variants including 18 pathogenic or variants of unknown significance were identified from 75 clinical samples and known variants of the reference samples were confirmed by Sanger sequencing and/or NGS. One false-negative results were present for Ion PGM/OTG-snpcaller for an indel variant misidentified as a single nucleotide variant. However, eight discordant results were present for Ion PGM Dx/Torrent Suite with both false-positive and -negative results. A 40-bp deletion, a 4-bp deletion and a 1-bp deletion variant was not called and a false-positive deletion was identified. Four other variants were misidentified as another variant. CONCLUSION: Ion PGM/OTG-snpcaller showed acceptable performance with good concordance with Sanger sequencing. However, Ion PGM Dx/Torrent Suite showed many discrepant results not suitable for use in a clinical laboratory, requiring further optimization of the data analysis for calling variants.

Dysregulation of Telomere Lengths and Telomerase Activity in Myelodysplastic Syndrome

BACKGROUND: Telomere shortening is thought to be involved in the pathophysiology of myeloid malignancies, but telomere lengths (TL) during interphase and metaphase in hematopoietic malignancies have not been analyzed. We aimed to assess the TLs of interphase and metaphase cells of MDS and telomerase activity (TA) and to find out prognostic significances of TL and TA. METHODS: The prognostic significance of TA by quantitative PCR and TL by quantitative fluorescence in situ hybridization (QFISH) of interphase nuclei and metaphase chromosome arms of bone marrow cells from patients with MDS were evaluated. RESULTS: MDS patients had shorter interphase TL than normal healthy donors (P<0.001). Average interphase and metaphase TL were inversely correlated (P=0.013, p arm; P=0.029, q arm), but there was no statistically significant correlation between TA and TL (P=0.258). The progression free survival was significantly shorter in patients with high TA, but the overall survival was not different according to average TA or interphase TL groups. Multivariable Cox analysis showed that old age, higher International Prognostic Scoring System (IPSS) subtypes, transformation to AML, no history of hematopoietic stem cell transplantation and short average interphase TL (<433 TL) as independent prognostic factors for poorer survival (P=0.003, 0.001, 0.005, 0.005, and 0.013, respectively). CONCLUSIONS: The lack of correlation between age and TL, TA, and TL, and the inverse relationship between TL and TA in MDS patients reflect the dysregulation of telomere status and proliferation. As a prognostic marker for leukemia progression, TA may be considered, and since interphase TL has the advantage of automated measurement by QFISH, it may be used as a prognostic marker for survival in MDS.

Utilization Analysis of Neonatal Red Blood Cell Aliquots and Development of Electronic Issuing System / 대한수혈학회지

BACKGROUND: Although transfusion in neonates needs to be strictly regulated due to the vulnerability of neonates, there is lack of systematic studies and the working process is not well-established. This study was aimed to point out the problems of current status and to improve the efficiency of systems used in blood aliquots for neonatal transfusions. METHODS: Total red blood cell (RBC) aliquots were analyzed between May 2009 and January 2016 in the neonate intensive care unit. We investigated the aliquot number, issued day interval from the first issued aliquot among the post-aliquots, patients' blood type, and discarded RBC units among the requested RBC units. RESULTS: Of the 472 RBC aliquots, 95.4% (450/472) were divided into two units. The distribution of patients' blood type was similar to that of the Korean population, in decreasing order: A blood group (34.3%), B group (28.2%), and O group (27.5%). The second, third, and forth units of post-aliquots were taken after an average of 49.9 (0∼617.9) hours. Among the post-aliquots, the number of units discarded without use was 22.5%. CONCLUSION: According to the evaluation of current status for neonatal transfusions, we should use aliquot RBC properly and reduce unnecessary requests for aliquot RBC. In addition, in order to reduce the number of near misses, we propose a new label to be attached on the aliquotted blood bags and suggest a development of electronic blood issuing system.

Bone Marrow Recurrence with Microscopic Pulmonary Tumor Embolism after Excision of Extramammary Paget's Disease in Scrotum Three Years Ago

Here we report a case of a 72-year-old male patient recurred in bone marrow alone with pulmonary tumor embolism after an excision of extramammary Paget's disease of scrotum 3 years ago. The patient received paclitaxel/carboplatin chemotherapy with respiratory support in intensive care unit. Four days after chemotherapy, the oxygen demand decreased and the patient was transferred to general ward. The platelet count recovered after 2 weeks. Finally, he died of hepatic failure from Paget's disease hepatic involvement confirmed by liver biopsy at 10 months after recurrence. This is a rare case of recurred extramammary Paget's disease in bone marrow alone with pulmonary tumor embolism, which was properly diagnosed with high suspicion and was successfully treated with immediate chemotherapy.

MALDI-TOF MS: Its Application in the Clinical Laboratory and a Paradigm Shift in Clinical Microbiology

In the past decade, clinical microbiology underwent revolutionary changes in methods used to identify microorganisms, a transition from slow and traditional microbial identification algorithms to rapid molecular methods and mass spectrometry (MS). Earlier, MS was clinically used as a highly complex method that was adapted for protein-centered analysis of samples in chemistry laboratories. Recently, a paradigm-shift happened when matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) MS was implemented to be used in microbiology laboratories for rapid and robust methods for accurate microbial identification. Two instrument platforms, marketed by well-established manufacturers, are beginning to displace automated phenotypic identification instruments and in some cases even genetic sequence-based identification practices. This review summarizes the current role of MALDI-TOF MS in clinical research, in diagnostic clinical microbiology laboratories, and serves as an introduction to MALDI-TOF MS, highlighting research associated with sample preparation, algorithms, interpretations, and limitations. Currently available MALDI-TOF MS instruments as well as software platforms that support the use of MALDI-TOF with direct specimens have been discussed in this review. Finally, clinical laboratories are consistently striving to extend the potential of these new methods, often in partnership with developmental scientists, resulting in novel technologies, such as MALDI-TOF MS, which could shape and define the diagnostic landscape for years to come.

Adverse Prognostic Impact of Bone Marrow Microvessel Density in Multiple Myeloma

BACKGROUND: Angiogenesis is important for the proliferation and survival of multiple myeloma (MM) cells. Bone marrow (BM) microvessel density (MVD) is a useful marker of angiogenesis and is determined by immunohistochemical staining with anti-CD34 antibody. This study investigated the prognostic impact of MVD and demonstrated the relationship between MVD and previously mentioned prognostic factors in patients with MM. METHODS: The study included 107 patients with MM. MVD was assessed at initial diagnosis in a blinded manner by two hematopathologists who examined three CD34-positive hot spots per patient and counted the number of vessels in BM samples. Patients were divided into three groups according to MVD tertiles. Cumulative progression-free survival (PFS) and overall survival (OS) curves, calculated by using Kaplan-Meier method, were compared among the three groups. Prognostic impact of MVD was assessed by calculating Cox proportional hazard ratio (HR). RESULTS: Median MVDs in the three groups were 16.8, 33.9, and 54.7. MVDs were correlated with other prognostic factors, including beta2-microglobulin concentration, plasma cell percentage in the BM, and cancer stage according to the International Staging System. Multivariate Cox regression analysis showed that high MVD was an independent predictor of PFS (HR=2.57; 95% confidence interval, 1.22-5.42; P=0.013). PFS was significantly lower in the high MVD group than in the low MVD group (P=0.025). However, no difference was observed in the OS (P=0.428). CONCLUSIONS: Increased BM MVD is a marker of poor prognosis in patients newly diagnosed with MM. BM MVD should be assessed at the initial diagnosis of MM.

Genotyping for RhD and RhCEce Antigens Using Free Circulating Nucleic Acids in Plasma and Serum / 대한수혈학회지

BACKGROUND: The Rh blood group includes several antigens, of which D, C, E, c, and e are clinically important. Although nucleic acids from whole blood can be used for Rh blood group genotyping, it is also possible to genotype free circulating fetal nucleic acids from plasma and serum. We performed Rh blood group phenotyping and genotyping using nucleic acids from whole blood and free circulating nucleic acids from plasma and serum, respectively. The results were compared. METHODS: Forty-four blood samples were phenotyped and genotyped for RhD and RhCE blood groups. Phenotyping was performed by hemagglutination assay. Further tests were performed on RhD-negative samples. Nucleic acids were extracted from whole blood, plasma, and serum. Plasma and serum were prepared after filtration and genotyped by real-time polymerase chain reaction. RESULTS: RhD blood group results showed one (2.3%) discrepant case in which the DEL phenotype appeared wild RHD genotype. Among nucleic acids, there were seven discrepant results: two from plasma and five from serum based on whole blood nucleic acids. RhCE blood group results showed three (6.8%) phenotype-genotype discordances. Among nucleic acids, seven (15.9%mpared to phenotypes. Kappa coefficients of serum were lower than those of plasma. CONCLUSION: RHD and RHCE genotype could be identified by assaying free circulating nucleic acids in plasma or serum. This study suggests that plasma is more reliable than serum as a specimen for RHD and RHCE genotyping of free circulating nucleic acids.

Probability of Obtaining Specific Antigen-Negative Blood Units through Antigen Testing in Korean Medical Institutes / 대한수혈학회지

BACKGROUND: When unexpected antibodies are identified, selection for specific antigen-negative blood units is needed in order to ensure transfusion safety. We estimated the number of blood units required for antigen testing to obtain specific antigen-negative units in Korean medical institutes. METHODS: We analyzed cases of selection for specific antigen-negative units for recipients who had antibodies identified in Seoul National University Bundang hospital from January 2008 to December 2010 and cases entered into the KRBP (Korean Rare Blood Program) online database from July 2013 to February 2014 from eight medical institutes. RESULTS: A total of 559 cases of 266 patients were analyzed. The antigen types requiring two units on average for one specific antigen-negative unit were E, P1, and Lea. Three units on average were required for one Fyb-negative blood unit, four units for one Jka-negative unit, four units for one Jkb-negative unit, 4.5 units for one Leb-negative unit, five units for one C-negative unit, six units for one M-negative unit, and seven units for one S-negative unit. In cases of obtaining specific antigen-negative units for more than one antigen type, three units on average were required for one E, c-negative unit and seven units for one C, e-negative unit. Other multiple antigen-negative units required up to 20 units. CONCLUSION: The accurate antigen-negative frequency in the Korean population should be investigated. Following this effort, the number of blood units required for selection of specific antigen-negative units could be predicted and practical measures for obtaining specific antigen-negative blood units could be suggested for Korean medical institutes.

Frequency of CD36 Deficiency on Platelets and Monocytes in Koreans / 대한수혈학회지

BACKGROUND: CD36 deficiency was first identified in a patient who showed refractoriness to HLA-matched platelet transfusion. CD36 deficiency can be divided into two subgroups. The type I phenotype is characterized by platelets and monocytes exhibiting CD36 deficiency. The type II phenotype lacks surface expression of CD36 in platelets only. In this study, the frequency of type I and type II CD36 deficiency in Koreans was evaluated. METHODS: A total of 220 samples were randomly selected from subjects who requested CBC testing from August 2013 to February 2014. The expression levels of CD36 on platelets and monocytes were analyzed by flow cytometry using FITC-conjugated CD36 antibodies. Correlation between the median fluorescence intensity of CD36 and the number of platelets or monocytes was evaluated using Pearson's correlation coefficient. RESULTS: Type I phenotype, lacking CD36 on platelets and monocytes, was present in 0.9% and type II, lacking CD36 on platelets, was present in 3.2%. The median fluorescence intensity of CD36 did not show correlation with the count of platelets or monocytes. CONCLUSION: Type I subjects may produce alloantibodies against CD36 following transfusion or pregnancy, leading to refractoriness to HLA-matched platelet transfusion, post-transfusion purpura, or neonatal immune thrombocytopenia. Studies to determine exact frequency of CD36 deficiency in Koreans, including a larger population, should be conducted, and more case reports on patients immunized against CD36 are also needed in order to elucidate the clinical importance and relevance of CD36 deficiency testing and the transfusion of CD36-deficient platelets.

The Current Status of Management for Specific Antigen-negative Blood Units in Korea / 대한수혈학회지

BACKGROUND: For pretransfusion testing, ABO and D antigen tests along with unexpected antibody screening tests are performed. When unexpected antibodies are identified, selection for specific antigen-negative blood units is needed in order to ensure safety of transfusion. METHODS: A questionnaire survey was conducted from August 23 to September 10, 2012 in 36 medical institutes in order to evaluate the current status of management for specific antigen-negative blood units in Korea. The questionnaire consisted of a method for detection of unexpected antibodies, the number of antibodies identified in the last year, and the antigen tests performed for specific antigen-negative blood units. For the institutes where blood donations are obtained, we asked about the enrollment of donors for specific antigen-negative or rare blood types. RESULTS: Among the 36 institutes, antigen testing for specific antigen-negative blood units was performed in 20 institutes. Of the remaining 15 institutes, except for one institute which answered as not applicable, eight institutes requested blood units at blood centers and another seven institutes replaced antigen tests with crossmatching tests. Among the 21 institutes where blood donations are obtained, two institutes had enrolled donors for specific antigen-negative or rare blood types. CONCLUSION: For selection of specific antigen-negative blood units for recipients who have identified antibodies, standardization of antibody detection tests and antigen tests is needed. In addition, the accurate antigen frequency in the Korean population should be investigated and donors for specific antigen-negative or rare blood types should be enrolled and managed systematically. Following these efforts, practical measures for obtaining specific antigennegative blood units could be suggested for medical institutes in Korea.

The application of an in situ karyotyping technique for mesenchymal stromal cells: a validation and comparison study with classical G-banding

The cytogenetic analysis of mesenchymal stromal cells (MSCs) is essential for verifying the safety and stability of MSCs. An in situ technique, which uses cells grown on coverslips for karyotyping and minimizes cell manipulation, is the standard protocol for the chromosome analysis of amniotic fluids. Therefore, we applied the in situ karyotyping technique in MSCs and compared the quality of metaphases and karyotyping results with classical G-banding and chromosomal abnormalities with fluorescence in situ hybridization (FISH). Human adipose- and umbilical cord-derived MSC cell lines (American Type Culture Collection PCS-500-011, PCS-500-010) were used for evaluation. The quality of metaphases was assessed by analyzing the chromosome numbers in each metaphase, the overlaps of chromosomes and the mean length of chromosome 1. FISH was performed in the interphase nuclei of MSCs for 6q, 7q and 17q abnormalities and for the enumeration of chromosomes via oligo-FISH in adipose-derived MSCs. The number of chromosomes in each metaphase was more variable in classical G-banding. The overlap of chromosomes and the mean length of chromosome 1 as observed via in situ karyotyping were comparable to those of classical G-banding (P=0.218 and 0.674, respectively). Classical G-banding and in situ karyotyping by two personnel showed normal karyotypes for both cell lines in five passages. No numerical or structural chromosomal abnormalities were found by the interphase-FISH. In situ karyotyping showed equivalent karyotype results, and the quality of the metaphases was not inferior to classical G-banding. Thus, in situ karyotyping with minimized cell manipulation and the use of less cells would be useful for karyotyping MSCs.

Adoption of Donor Screening Policy in a Tissue Bank at a Tertiary Hospital / 대한수혈학회지

BACKGROUND: Tissues for transplantation can save lives or restore essential functions. According to national policies and regulations, access to suitable transplantation, as well as the level of safety, quality, efficacy of donation, and transplantation of tissues, differ significantly between countries. We reviewed a few guidelines on tissue banking from the aspect of screening tests. In addition, four-year experience with screening panels for donated bones and donors at a tertiary hospital is introduced. METHODS: Seven national and international guidelines for screening tests for donors and donated tissues were reviewed. At our institution, screening tests for donation involve two steps. At retrieval, the first screening panel, including ABO/Rh typing, unexpected antibody screening, VDRL, HBsAg, anti-HBs, anti-HBc IgM, anti-HCV, anti-HIV, and microbiological cultures was performed. The second screening panel, including the same tests, except culture studies, was performed after 90 days. From 2008 to 2011, a total of 245 retrievals of bone tissue were performed and the screening panel results were analyzed. RESULTS: Mandatory screening serologic tests for living donors can differ according to local law or regulation and/or screening for endemic diseases. At our institution, among 245 donated bones for a period of four years, 61 bone tissues were discarded due to noncompliance for the second screening (n=32), contamination or no culture study results (n=9), abnormal serologic test results (n=8), and so on. CONCLUSION: Donor screening policies for tissue banking are various according to national laws or endemic disease status. Second screening tests with consideration of the window period should be adopted.

Human Platelet Antigen Genotyping Using a Multiplex Single-Base Primer Extension Reaction in Koreans / 대한수혈학회지

BACKGROUND: Alloimmunization of human platelet antigens (HPA) is associated with clinically significant disease, such as platelet refractoriness, neonatal alloimmune thrombocytopenia, or posttransfusion purpura. It is determined by single nucleotide polymorphism of genes for platelet membrane glycoprotein. To date, approximately 27 HPAs have been discovered, and their frequencies differ depending on ethnicity and country. METHODS: We conducted an investigation of prevalence of HPA in the Korean population using a multiplex single-base primer extension reaction (SNaPshot). With 84 specimens from healthy donors, HPA genotyping was performed on 11 different HPAs, including HPA-1, -2, -3, -4, -5, -6, -7, -8, -9, -13, and -15. RESULTS: A total of 90 blood samples were genotyped. The genotype frequencies of HPA were as follows: HPA-1a/1a: 100.0%, -2a/2a: 83.3%, -2a/2b: 14.3%, -2b/2b: 2.4%, -3a/3a: 39.3%, -3a/3b: 52.4%, -3b/3b: 8.3%, -4a/4a: 100.0%, -5a/5a: 95.2%, -5a/5b: 4.8%, -6a/6a: 94.0%, -6a/6b: 6.0%, -7a/7a: 100.0%, -8a/8a: 100.0%, -9a/9a: 97.6%, -9a/9b: 2.4%, -13a/13a: 100.0%, -15a/15a: 23.8%, -15a/15b: 51.2%, and -15b/15b: 25.0%. CONCLUSION: The SNaPshot assay was employed for detection of SNPs in various clinically significant HPA genes. In addition to well-known frequencies of previously reported HPA-1 to -8, this study showed frequencies of HPA-9, -13, and -15 in Koreans for the first time. The SNaPshot technique might be suitable for use in actual clinical testing in patients with platelet alloimmunization.

Analysis of Xenotropic Murine Leukemia Virus-Related Virus (XMRV) in Korean Blood Donors in a Medical Center / 대한수혈학회지

BACKGROUND: Xenotropic murine leukemia virus-related virus (XMRV) has been detected in peripheral blood mononuclear cells (PBMNs), therefore, it has been regarded as being infectious and transmittable by transfusion. Thus, we attempted to detect XMRV in blood samples in order to confirm the absence of XMRV from blood donors. METHODS: We achieved 165 blood donors and four chronic fatigue syndrome (CFS) patients. We performed real-time polymerase chain reaction using the LightCycler 480 (Roche, Penzberg, Germany) for the gag and env genes of the XMRV genome. DNA was extracted from peripheral blood samples. We used Uracil-N-Glycosylase in order to prevent contamination and DNA extracted from mouse embryonic fibroblasts (MEF) for amplification control. RESULTS: No XMRV was detected in any of the blood donors in both the gag and env genes. In four CFS patients, amplification was not detected in the gag gene. In two of four CFS patients, amplifications were detected and the melting temperature was in agreement with that of MEF control in the env gene. CONCLUSION: Although XMRV was not present in blood samples from blood donors, this is the first report on XMRV in Korean blood donors. We confirmed the absence of XMRV in Korean blood donors, the same as studies reported in other countries.

Effects of granulocyte-colony stimulating factor and the expression of its receptor on various malignant cells / 대한혈액학회지

BACKGROUND: Granulocyte-colony stimulating factor (G-CSF) is extensively used to improve neutrophil count during anti-cancer chemotherapy. We investigated the effects of G-CSF on several leukemic cell lines and screened for the expression of the G-CSF receptor (G-CSFR) in various malignant cells. METHODS: We examined the effects of the most commonly used commercial forms of G-CSF (glycosylated lenograstim and nonglycosylated filgrastim) on various leukemic cell lines by flow cytometry. Moreover, we screened for the expression of G-CSFR mRNA in 38 solid tumor cell lines by using real-time PCR. RESULTS: G-CSF stimulated proliferation (40-80% increase in proliferation in treated cells as compared to that in control cells) in 3 leukemic cell lines and induced differentiation of AML1/ETO+ leukemic cells. Among the 38 solid tumor cell lines, 5 cell lines (hepatoblastoma, 2 breast carcinoma, squamous cell carcinoma of the larynx, and melanoma cell lines) showed G-CSFR mRNA expression. CONCLUSION: The results of the present study show that therapeutic G-CSF might stimulate the proliferation and differentiation of malignant cells with G-CSFR expression, suggesting that prescreening for G-CSFR expression in primary tumor cells may be necessary before using G-CSF for treatment.