BACKGROUND: One of the major concerns with biobanking is the absence of standard operating procedures to eliminate pre-analytical variation arising from sample collection, preparation, and storage. Currently, there is a lack of tools to carry out quality control procedures for stored blood samples. The aim of this study is to assess the quality of stored blood samples in our biobank and to suggest appropriate indicators for their quality control. METHODS: The stored blood samples that we tested have been registered into our biobank since 2003. These were transferred to our biobank after carrying out routine requested tests, because the samples would have otherwise been discarded. For the purpose of quality control, we analyzed the concentrations and the integrity of DNA and RNA extracted from the stored samples and tested the levels of several serum proteins; the results were compared with the corresponding pre-storage levels. RESULTS: A total of 19 samples were stored from 2006 to 2009. Of the 22 samples stored between 2003 and 2005, 50% showed complete DNA integrity. However, sufficient RNA integrity was noted in only 1 sample stored as recently as 2009. High blood urea nitrogen levels were also noted in the stored sera, but the increase did not correlate to the duration of storage. CONCLUSIONS: The amount and integrity of nucleic acids extracted from stored blood samples are potential indicators that can be used for quality control. A guideline for the quality assessment of stored blood samples in a biobank is urgently needed.
Blood Banks/*standards ; Blood Proteins/chemistry/standards ; Blood Urea Nitrogen ; DNA/*analysis/chemistry/standards ; Laboratory Techniques and Procedures ; Quality Control ; RNA/*analysis/chemistry/standards ; Specimen Handling/methods

BACKGROUND: Autoanalyzer ADVIA2120 uses intracellular peroxidase concentration to perform white blood cell (WBC) differential. Therefore, in specimens containing neutrophils with low peroxidase concentration, neutrophils can be miscounted as monocytes or large unstained cells resulting in pseudoneutropenia. Myeloperoxidase deficiency can be detected by the mean peroxidase index (MPXI). The aims of this study are to establish the reference interval of MPXI and define a cut off value for manual slide review to discriminate pseudoneutropenia. METHODS: We calculated reference intervals as mean+/-2SD according to the indirect method of CLSI C28-A3 guideline from MPXI data of 5,802 individuals who took routine health checkup from April 2010 to June 2012. We performed manual slide review on specimens with low MPXI and compared neutrophil differential count of manual method with that of autoanalyzer. When neutrophil differential in manual slide review was >20%P higher than autoanalyzer result, it was regarded as a pseudoneutropenia. We performed ROC analysis using the MPXI results of samples with and without pseudoneutropenia to define a cutoff to discriminate pseudoneutropenia. RESULTS: The reference intervals of MPXI in total population, male, and female were -4.9 to 7.5, -5.5 to 7.3, and -4.5 to 7.5, respectively. The mean value of MPXI was significantly higher in female than in male and there was no difference by age. Twenty-two pseudoneutropenia samples from 7 patients were identified. ROC analysis yielded cutoff value of -20.7 with 94.9% of sensitivity and 77.3% of specificity. CONCLUSIONS: MPXI may be used in the manual slide review guideline for detecting pseudoneutropenia.
Discrimination (Psychology) ; Female ; Humans ; Leukocytes ; Male ; Metabolism, Inborn Errors ; Monocytes ; Neutrophils ; Peroxidase ; ROC Curve

BACKGROUND: Screening for healthy blood donors through donor interviews is essential to the safety of donors and blood resources. Our goal was to suggest educational material for donor interviewers and donors, as well as supplemental material for interview sites, which will help provide an effective interview process. METHODS: We conducted surveys regarding experiences in donor interviews from donor interviewers and cognitive interviews about difficulties during interview from blood donors between September and October of 2015. We additionally conducted a post-survey about provided educational and supplemental materials between December 2015 and January 2016. RESULTS: The possibility of an incorrect answer in the donor history questionnaire (DHQ) was high for questions about sexual contact, imprisonment, or medication, and the reasons were incorrect memories, ignorance about donor interview, or protection of privacy. Cognitive interviews of donors revealed questions and terminology that are difficult to understand. Donor interviewers could obtain improved understanding of the DHQ through educational materials, which were found to be useful for new interviewers or donors. Use of a supplemental flip book for the interview process was found to be useful, especially for blood centers with small blood donations. CONCLUSION: This study investigated difficulties in the donor interview from the perspective of donor interviewers and donors and suggested educational and supplemental materials to address these difficulties. These materials will induce correct and honest answers from blood donors through education and guidance about the donor interview process and help secure the safety of blood products.
Blood Donors* ; Cognition ; Education ; Humans ; Mass Screening ; Privacy ; Tissue Donors*

To evaluate the current status of cerebrovascular disease in Korea, we performed a retrospective study on 1260 cases of stroke who had been admitted to or had visited Seoul National University Hospital, Choongnam National University Hospital, Hanyang University Hosipital, and Chung Ang University Hospital in the period from Jul 1, 1986 to Jun 30, 1987. The incidence was highest in sixth and seventh decades. Ischemic stroke (51.1%) was more common thar, hemorrhagic stroke (47.6%), and thrombotic infarction was the leading type (37.7%) of all kinds of strokes. Middle cerebral arterial territory was the most commonly involved site in thrombotic and embolic stroke. Among intracerebral hemorrhage, basal ganglia hemorrhage was found in 47.7%, and was followed by thalamic(23.8%), lobar (15.3%) and pontine hemorrhage (9.4%). Primary intraventricular hemorrhage (IVH) was relatively rare (8 cases, 7.5% of all IVH), and hemorrhages in basal ganglia or thalamus frequently induced secondary IVH. 16 cases (10.7%) of subarachnoid hamorrhage were accompanied by secondary IVH. This study suggests changing trends of stroke in Korea, and multicenter prospective study using stroke registry is requird for confirmation.
Basal Ganglia ; Basal Ganglia Hemorrhage ; Cerebral Hemorrhage ; Hemorrhage ; Incidence ; Infarction ; Korea* ; Retrospective Studies ; Seoul ; Stroke ; Thalamus

BACKGROUND: Urinalysis is one of the most commonly performed tests in clinical laboratories. In this study, we compared YD URiSCAN PluScope (PluScope; YD Diagnostics Corp., Korea) and Sysmex UF-1000i (UF-1000i; Sysmex Corp., Japan) for urine microscopic sediment analysis. METHODS: A total of 404 fresh urine samples were collected and analyzed using PluScope, UF-1000i, and manual microscopy. Quantitative correlation analyses for red blood cells (RBCs), white blood cells (WBCs), epithelial cells (EC), and casts were performed using Spearman's correlation. We evaluated agreement among the three systems by using weighted Cohen's κ and calculating concordance rates within one grade of difference for semiquantitative and qualitative parameters. RESULTS: There were moderate-high correlations between PluScope and UF-1000i for RBCs, WBCs, and ECs (r=0.542, 0.714, and 0.571, respectively) but negligible correlation for casts (r=0.186). There were moderate-high correlations between manual microscopy and automated devices for RBCs, WBCs, and ECs (r=0.550–0.745) but negligible correlations for casts (PluScope: r=0.247; UF-1000i: r=0.223). The pairwise concordance rates within one grade difference among the three methods were good for RBCs, WBCs, and ECs (95.0%–99.0%, κ=0.41–0.74). For casts, the concordance rate between PluScope and manual microscopy was fair (96.8%, κ=0.25), but concordance rates between UF-1000i and manual microscopy and between the two automated devices were poor (81.2% and 81.7%; κ=0.04 and 0.06, respectively). CONCLUSIONS: The two automated urine sediment analyzers showed a moderate-high correlation and concordance rate. They showed good correlations and concordance rates for RBCs, WBCs, and ECs. However, manual microscopic examinations are still needed for reviewing and confirming the presence of pathologic particles in urine, such as casts and crystals.
Epithelial Cells ; Erythrocytes ; Flow Cytometry* ; Leukocytes ; Microscopy ; Urinalysis

BACKGROUND: Since free light chain (FLC) is metabolized in the kidney, serum FLC concentration and kappa/lambda ratio are increased in patients with decreased renal function, even in the absence of monoclonal protein. In this study, we measured serum FLC levels to investigate the change in kappa/lambda ratios in relation to the severity of renal dysfunction. METHODS: Serum FLC concentrations were measured in 92 archived serum samples from patients diagnosed with chronic kidney disease using the Freelite assay (The Binding Site Group Ltd., UK), and kappa/lambda ratios were calculated. Serum creatinine levels were assayed to calculate estimated glomerular filtration rate (eGFR), and patients were divided into subgroups according to Kidney Disease Improving Global Outcomes (KDIGO) guidelines. We analyzed the difference in serum FLC levels and kappa/lambda ratios between subgroups. RESULTS: Serum FLC levels and kappa/lambda ratios increased depending on the severity of renal dysfunction. When patients were classified by setting cut-off value of eGFR as 60 mL/min/1.73 m2 (group A: eGFR ≥60 mL/min/1.73 m2, group B: < 60 mL/min/1.73 m2), the kappa/lambda ratio of group B was significantly higher than that of group A (group B: 1.60±0.46 vs. group A: 1.35±0.27, P=0.018). Serum FLC kappa/lambda ratios were within the previously determined renal reference interval (0.37–3.1). CONCLUSIONS: When interpreting results of serum FLC kappa/lambda ratio, renal function status should be considered in addition to hematological findings. If renal function deteriorates, a wider renal reference interval is preferred instead of the usual reference interval.
Binding Sites ; Creatinine ; Glomerular Filtration Rate ; Humans ; Kidney ; Kidney Diseases ; Renal Insufficiency, Chronic

Background: Intraoperative measurement of intact parathyroid hormone (iPTH) levels is crucial for confirming complete removal of hyperfunctioning parathyroid glands during parathyroidectomy and for detecting parathyroid gland damage during thyroidectomy. The use of plasma samples can shorten the turnaround time (TAT) for iPTH. The present study explored the effectiveness of using plasma samples for iPTH quantitation by comparison with the corresponding serum samples. We also evaluated the analytical performance of iPTH. Methods: The TAT of plasma and serum samples analyzed in March 2019 was compared. In addition, comparative evaluation of the iPTH levels in 100 paired plasma and serum samples were performed. Analytical performances including within-run and within-laboratory precision, and linearity were evaluated in plasma samples using the ADVIA Centaur iPTH assay (Siemens Healthineers, Germany). The reference range was verified with plasma samples collected from 20 healthy adults. Results: Plasma iPTH tests showed shorter TAT values (P<0.001) and higher iPTH levels (P<0.001) than serum. Correlation analysis between plasma and serum iPTH levels showed a strong positive correlation (r=0.925). The within-run and within-laboratory precision values were within the manufacturer's recommendation. iPTH showed linearity from 5.1 to 1,670.0 pg/mL (R2=0.999). The plasma iPTH levels from 20 healthy adults were within the reference range, thus validating our method. Conclusions The plasma iPTH levels were higher than the serum levels, with a strong positive correlation. The TAT of plasma samples was considerably shorter than that of serum. iPTH quantitation from plasma samples is preferable when rapid results are required.

BACKGROUND: BRCA1 (breast-cancer gene 1) is a tumor suppressor gene that accounts for nearly all families of both early onset breast and ovarian cancer and about 45% of families with breast cancer only. Sporadic nonhereditary breast cancer is recognized as the most common form of this malignancy. However, presence of germ-line mutations in the BRCA1 gene of these tumors is an infrequent event. The BRCA1 protein includes a ring domain and an acidic domain, both of which are characteristics of certain transcription factors, as well as two putative nuclear localization signals (NLS) that interact with importin-alpha. The normal BRCA1 protein is located in the nucleus of most breast-cell types whereas the BRCA1 protein of breast cancer cells is aberrantly localized in the cytoplasm. This mislocation of the BRCA1 protein in breast cancer cells may be due to defects in the NLS receptor-mediated pathway for the nuclear import of the BRCA1 gene product. Identification of importin-alpha mutations as a cellular protein responsible for the nuclear import of BRCA1 in breast-cancer cell lines and primary breast cancers is the focus of this investigation. METHODS: A series of 15 surgical samples of breast cancer and 3 samples of breast-cancer cell lines (Hs578T, ZR75-1, MCF-7) was assayed for the presence of the deletion mutant in importin-alpha by using both RT-PCR amplification of importin-alpha transcripts and sequencing analysis. RESULTS: Three of the 15 primary breast cancers and 1 of the 3 breast-cancer cell lines showing deletions in importin-alpha transcripts produced two different truncated transcripts. 1208 bp deletions were observed in transcripts from breast cancer (T-1, T-3) and ZR75-1, which is specified by the nucleotide 251-1458 of the transcript. Another transcript encoded by primary breast cancer (T-2) included a 1312 bp deletion in the nucleotide 61-1372 of the transcript. CONCLUSIONS: The deletions eliminated part of the importin-alpha transcript segment encoding the putative NLS-binding domain but not the importin-beta binding domain, suggesting that these deletion mutants could not bind to NLS of the BRCA1 protein. These results suggest that the composite effects of mislocationof the BRCA1 protein by deletion of the NLS-binding domain in importin-alpha may contribute to tumorigenesis in sporadic breast cancer.
Active Transport, Cell Nucleus ; alpha Karyopherins* ; Alternative Splicing ; beta Karyopherins ; BRCA1 Protein ; Breast Neoplasms* ; Breast* ; Carcinogenesis ; Cell Line ; Cytoplasm ; Genes, BRCA1 ; Genes, Tumor Suppressor ; Germ-Line Mutation ; Humans ; Nuclear Localization Signals ; Ovarian Neoplasms ; Transcription Factors

Alpha-thalassemia (alpha-thalassemia), which is prevalent in the Mediterranean region, is caused by deficient synthesis of the alpha-globin chains. It is commonly caused by HBA1 and/or HBA2 gene deletion and is diagnosed by DNA sequence analysis. The proband was a 38-year-old woman who was found to have microcytic and hypochromic anemia on a routine health checkup. Results of the Hb electrophoresis (EP) and direct sequencing of the HBA1 and HBA2 genes were found to be normal. As multiplex ligation-dependent probe amplification (MLPA) for the HBA1 and HBA2 genes revealed heterozygous deletion, she was diagnosed with heterozygous alpha+-thalassemia. Although routine laboratory tests revealed similar findings in the proband's father, brother and niece, MLPA revealed heterozygous deletions of the HBA1 or HBA2 gene in her brother and niece. In summary, we report a case of heterozygous alpha+-thalassemia in a Korean family that was detected by MLPA. We recommend that patients with suspected hemoglobinopathies should be followed-up further with MLPA, especially when Hb EP shows a normal pattern.
alpha-Globins ; alpha-Thalassemia ; Anemia, Hypochromic ; Electrophoresis ; Fathers ; Female ; Gene Deletion ; Hemoglobin A, Glycosylated ; Hemoglobinopathies ; Humans ; Mediterranean Region ; Multiplex Polymerase Chain Reaction ; Sequence Analysis, DNA ; Siblings

PURPOSE: Previous studies and our own clinical experience suggest that concurrent corticosteroid treatment for severe rickettsial disease with multiorgan failure may improve the clinical course or reduce mortality. However, the use of corticosteroids as adjunctive treatment for rickettsial diseases is controversial. We attempted to determine the influences of corticosteroid on the growth of Orientia tsutsugamushi in vitro to justify and evaluate the clinical applicability of corticosteroid in rickettsial disease. MATERIALS AND METHODS: L929 cells were infected with Orientia tsutsugamushi Gilliam. Dexamethasone was added to the cells at final concentrations of 10(1) and 10(7) pg/mL. Cultures were incubated at 35degrees C and processed for flow cytometry on the 6th day after addition of dexamethasone. RESULTS: Observation on the 6th day after treatment with dexamethasone in infected cultures revealed that there was no difference in fluorescence intensity among the treatment wells. Treatment of the cells with dexamethasone at concentrations of 10(1) and 10(7) pg/mL showed no influence on the growth of Orientia tsutsugamushi. CONCLUSION: Our results to show that isolated corticosteroid does not enhance the replication of Orientia tsutsugamushi in vitro. Concurrent use of anti-inflammatory or immunosuppressive doses of corticosteroids in conjunction with antibiotics may not have detrimental effects on the course of scrub typhus.
Animals ; Cell Line ; Cell Proliferation/drug effects ; Dexamethasone/*pharmacology ; Flow Cytometry ; Interferon-gamma/pharmacology ; Mice ; Orientia tsutsugamushi/*drug effects/growth & development ; Scrub Typhus/drug therapy/microbiology