Effect of Personalized Blue-Enriched White Light Intervention on Rest–Activity and Light Exposure Rhythms in Mild and Moderate Alzheimer’s Disease

Objective@#We aimed to examine the effectiveness of personalized light intervention using a blue-enriched light-emitting-diodes device on rest–activity rhythm (RAR) and light exposure rhythm (LER) in patients with mild and moderate Alzheimer’s disease (AD). @*Methods@#AD patients with poor sleep quality and/or insomnia symptoms were assigned into either an experimental group (EG) or control group (CG) in a single-blind design. Personalized light intervention was given at 9–10 h after individual dim light melatonin onset, lasting for 1 h every day for two weeks in the EG (77.36±5.79 years, n=14) and CG (78.10±7.98 years, n=10). Each patient of CG wore blue-attenuating sunglasses during the intervention. Actigraphy recording at home for 5 days was done at baseline (T0), immediate postintervention (T1), and at four weeks after intervention (T2). The variables of RAR and LER were derived using nonparametric analysis. @*Results@#We found a significant time effect on the intradaily variability (IV) of RAR at T2 with respect to T0 (p=0.039), indicating reduced IV of RAR at four weeks after personalized light intervention regardless of blue-enriched light intervention. There was a time effect on the IV of LER at T1 with respect to T0 (p=0.052), indicating a reduced tendency in the IV of LER immediately after intervention. @*Conclusion@#Our personalized light intervention, regardless of blue-enriched light source, could be useful in alleviating fragmentation of RAR and LER in AD patients.

Serotype Distribution and Antimicrobial Resistance of Invasive and Noninvasive Streptococcus pneumoniae Isolates in Korea between 2014 and 2016

BACKGROUND: Several factors contribute to differences in Streptococcus pneumoniae serotype distribution. We investigated the serotype distribution and antimicrobial resistance of S. pneumoniae isolated between 2014 and 2016 in Korea. METHODS: We collected a total of 1,855 S. pneumoniae isolates from 44 hospitals between May 2014 and May 2016, and analyzed the serotypes by sequential multiplex PCR. We investigated the distribution of each serotype by patient age, source of the clinical specimen, and antimicrobial resistance pattern. RESULTS: The most common serotypes were 11A (10.1%), followed by 19A (8.8%), 3 (8.5%), 34 (8.1%), 23A (7.3%), and 35B (6.2%). The major invasive serotypes were 3 (12.6%), 19A (7.8%), 34 (7.8%), 10A (6.8%), and 11A (6.8%). Serotypes 10A, 15B, 19A, and 12F were more common in patients ≤5 years old, while serotype 3 was more common in patients ≥65 years old compared with the other age groups. The coverage rates of pneumococcal conjugate vaccine (PCV)7, PCV10, PCV13, and pneumococcal polysaccharide vaccine 23 were 11.8%, 12.12%, 33.3%, and 53.6%, respectively. Of the 1,855 isolates, 857 (46.2%) were multi-drug resistant (MDR), with serotypes 11A and 19A predominant among the MDR strains. The resistance rates against penicillin, cefotaxime, and levofloxacin were 22.8%, 12.5%, and 9.4%, respectively. CONCLUSIONS: There were significant changes in the major S. pneumoniae serotypes in the community. Non-PCV13 serotypes increased in patients ≤5 years old following the introduction of national immunization programs with the 10- and 13-polyvalent vaccines.

Diagnostic Performance of %-2proPSA and Prostate Health Index for Prostate Cancer: Prospective, Multi-institutional Study

BACKGROUND: We evaluated the clinical performance of [-2]proPSA (p2PSA) and its derivatives in predicting the presence and aggressiveness of prostate cancer (PCa) in Korean men. METHODS: A total of 246 men with total prostate-specific antigen (tPSA) ≥ 3.5 ng/mL who underwent their first prostate biopsy were included in this prospective, multicenter, observational study. Diagnostic accuracy of tPSA, free-to-total PSA ratio (%fPSA), p2PSA, %p2PSA, and the Beckman Coulter prostate health index (PHI) was assessed by receiver operating characteristic curve analyses and logistic regression analyses. RESULTS: Overall, PCa was detected in 125 (50.8%) subjects. In men with tPSA 3.5–10 ng/mL, the detection rate of PCa was 39.4% (61/155). In this group, PHI and %p2PSA were the most accurate predictors of PCa and significantly outperformed tPSA and %fPSA; area under the curve for tPSA, %fPSA, %p2PSA, and PHI was 0.56, 0.69, 0.74, and 0.76, respectively. PHI was also the strongest predictor of PCa with Gleason score ≥ 7. CONCLUSION: This study demonstrates the superior clinical performance of %p2PSA and PHI in predicting the presence and aggressiveness of PCa in Korean men. The %p2PSA and PHI appear to improve detection of PCa and provide prognostic information.

Identification of 10 Candidate Biomarkers Distinguishing Tuberculous and Malignant Pleural Fluid by Proteomic Methods

PURPOSE: Pleural effusion, an accumulation of fluid in the pleural space, usually occurs in patients when the rate of fluid formation exceeds the rate of fluid removal. The differential diagnosis of tuberculous pleurisy and malignant pleural effusion is a difficult task in high tuberculous prevalence areas. The aim of the present study was to identify novel biomarkers for the diagnosis of pleural fluid using proteomics technology. MATERIALS AND METHODS: We used samples from five patients with transudative pleural effusions for internal standard, five patients with tuberculous pleurisy, and the same numbers of patients having malignant effusions were enrolled in the study. We analyzed the proteins in pleural fluid from patients using a technique that combined two-dimensional liquid-phase electrophoresis and matrix assisted laser desorption/ionization-time of flight-mass spectrometry. RESULTS: We identified a total of 10 proteins with statistical significance. Among 10 proteins, trasthyretin, haptoglobin, metastasis-associated protein 1, t-complex protein 1, and fibroblast growth factor-binding protein 1 were related with malignant pleural effusions and human ceruloplasmin, lysozyme precursor, gelsolin, clusterin C complement lysis inhibitor, and peroxirexdoxin 3 were expressed several times or more in tuberculous pleural effusions. CONCLUSION: Highly expressed proteins in malignant pleural effusion were associated with carcinogenesis and cell growth, and proteins associated with tuberculous pleural effusion played a role in the response to inflammation and fibrosis. These findings will aid in the development of novel diagnostic tools for tuberculous pleurisy and malignant pleural effusion of lung cancer.

Effects of Transglutaminase 2 Inhibition on Ventilator-Induced Lung Injury

This study was performed to examine the role of transglutaminase 2 (TG2) in ventilator-induced lung injury (VILI). C57BL/6 mice were divided into six experimental groups: 1) control group; 2) lipopolysaccharide (LPS) group; 3) lung protective ventilation (LPV) group; 4) VILI group; 5) VILI with cystamine, a TG2 inhibitor, pretreatment (Cyst+VILI) group; and 6) LPV with cystamine pretreatment (Cyst+LPV) group. Acute lung injury (ALI) score, TG2 activity and gene expression, inflammatory cytokines, and nuclear factor-kappaB (NF-kappaB) activity were measured. TG2 activity and gene expression were significantly increased in the VILI group (P < 0.05). Cystamine pretreatment significantly decreased TG2 activity and gene expression in the Cyst+VILI group (P < 0.05). Inflammatory cytokines were higher in the VILI group than in the LPS and LPV groups (P < 0.05), and significantly lower in the Cyst+VILI group than the VILI group (P < 0.05). NF-kappaB activity was increased in the VILI group compared with the LPS and LPV groups (P < 0.05), and significantly decreased in the Cyst+VILI group compared to the VILI group (P = 0.029). The ALI score of the Cyst+VILI group was lower than the VILI group, but the difference was not statistically significant (P = 0.105). These results suggest potential roles of TG2 in the pathogenesis of VILI.

Relationships of Coagulation Factor XIII Activity with Cell-Type and Stage of Non-Small Cell Lung Cancer

PURPOSE: Factor XIII (FXIII), a thrombin-activated plasma transglutaminase zymogen, is involved in cancer development and progression through a triggered coagulation pathway. The aim of this study was to examine whether FXIII activity levels differed in non-small cell lung cancer (NSCLC) patients according to histological types and TNM stage when compared with healthy subjects. MATERIALS AND METHODS: Twenty-eight NSCLC patients and 28 normal controls who had been individually age-, gender-, body mass index-, smoking status-, and smoking amount-matched were enrolled: 13 adenocarcinomas, 11 squamous cell carcinomas, and four undifferentiated NSCLCs; four stage I, two stage II, 12 stage III, and 10 stage IV NSCLCs. FXIII activity was measured using fluorescence-based protein arrays. RESULTS: The median FXIII activity level of the NSCLC group [24.2 Loewy U/mL, interquartile range (IQR) 14.9-40.4 Loewy U/mL] was significantly higher than that of the healthy group (17.5 Loewy U/mL, IQR 12.6-26.4 Loewy U/mL) (p=0.01). There were no differences in FXIII activity between adenocarcinoma (median 18.6 Loewy U/mL) and squamous cell carcinoma (median 28.7 Loewy U/mL). NSCLC stage significantly influenced FXIII activity (p=0.02). The FXIII activity of patients with stage III NSCLC (median 27.3 Loewy U/mL, IQR 19.3-40.5 Loewy U/mL) was significantly higher than those of patients with stage I or II (median 14.0 Loewy U/mL, IQR 13.1-23.1 Loewy U/mL, p=0.04). FXIII activity was negatively correlated with aPTT in NSCLC patients (r=-0.38, p=0.04). CONCLUSION: Patients with advanced-stage NSCLC exhibited higher coagulation FXIII activity than healthy controls and early-stage NSCLC patients.

Reduction of Influenza A (H1N1) Virus Through a Leukoreduction Filter / 대한수혈학회지

BACKGROUND: Leukoreduction can reduce the risk of HLA alloimmunization, recurrent febrile nonhemolytic transfusion reactions, and several transfusion-transmitted infectious diseases, including cytomegalovirus infection. Transmission of the new influenza A (H1N1) virus through transfusion may be a concern. We evaluated the effect of filtration with a leukoreduction filter on H1N1 genomes. METHODS: To evaluate the effect of filtration by a leukoreduction filter on H1N1 genomes, we analyzed pre- and post-filtered samples from nasopharyngeal swabs and 10 positive plasma samples using real time RT-PCR. RESULTS: The 10 samples (nasopharyngeal swabs and plasma) contained H1N1 RNA, and filtration with a leukoreduction filter reduced these levels (threshold cycle values from 31.42+/-2.06 to 38.84+/-1.47 in nasopharyngeal swabs, from 35.63+/-2.19 to 39.38+/-2.65 in plasma samples). CONCLUSION: Filtration with a leukoreduction filter can reduce H1N1 genome levels, but may not be completely sufficient for total eradication of this pathogen.

A Study to Verify the Stability of Currently Stored Blood Donor Samples (A First Year Study) / 대한수혈학회지

BACKGROUND: The Korean Red Cross (KRC) has stored blood donor samples for 10 years under -20degrees C since 2004. These samples have been used for investigating transfusion related infections and for Look-back studies. We designed an experimental scheme to verify the stability of stored blood samples. METHODS: We collected and prepared samples such as blood donor samples (HBV, HCV, HIV nucleic acid positive; n=90), the HIV infected patient samples (n=20), the WHO nucleic acid international standards serologic positive samples (HBsAg, anti-HCV, anti-HIV; n=120) and the negative samples (n=20). The samples were aliquoted in cryo tubes with volumes of 0.5~5 mL and they were stored at -20~-30degrees C and -70~-80degrees C. We used enzyme immunoassay, chemiluminescence immunoassay and quantitative PCR for the base line and the follow up studies. The linear mixed statistical model using SAS 9.1 for windows was used for statistical analysis. RESULTS: The results of the baseline test of the stored samples showed a variable range of viral load (10(1)~10(7) IU/mL or copies/mL) and optical density (S/CO 3.0~500). The results of the stored samples after 6 month (n=82) did not show any significant differences compared to the baseline data for the viral loads (P>0.05) and the qualitative serologic tests. CONCLUSION: We established an experimental scheme to verify the stability of the stored blood donor samples. From now on, the stability of the stored samples is going to be monitored by every 6 month for 10 years.

Protein Profile Changes in Platelet Concentrates According to Storage and Leukoreduction- Analysis Using Proteomics Technology / 대한진단검사의학회지

BACKGROUND: Knowing how the protein profile of platelet products changes with storage or leukoreduction may give us greater insight into cell physiology and the cause of transfusion reactions other than cytokines and chemokines. METHODS: We filtered four packs of platelet concentrates (PC) within 24 hr of blood collection and after 120 hrs of storage. Four aliquots of each supernatant in PC were obtained: pre-storage+prefiltration, pre-storage+post-filtration, post-storage+pre-filtration and post-storage+post-filtration. Routine chemistry tests and a two-dimensional electrophoresis (2-DE) were performed. The stained images were analyzed and the significant spots were identified using a peptide mass finger printing (PMF) with matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) analysis after trypsin digestion. RESULTS: The protein spots increased with storage and decreased after filtration (P<0.05, prestorage+post-filtration). The spot density of various proteins, including macrophage inflammatory protein-2 alpha, megakaryocyte colony stimulating factor and interleukin-22 changed with storage and leukoreduction. CONCLUSIONS: The database of identified protein spots and their changes produced in this study is a useful basic tool for future studies on the mechanism of transfusion reactions. Further studies should validate the significance of each protein spot.

Using Medical Error Cases for Patient Safety Education / 한국의학교육

PURPOSE: To draw attention to patient safety and increase its awareness among medical students, we developed a program that teaches patient safety based on common medical error cases. The aim of this study is to introduce this program and improve student receptivity to it. METHODS: As part of the "Patient, Doctor, and Society" course, third-year medical students participated in 8 hours of a medical error education program. Students discussed recent, typical medical lawsuits that were generated from internal medicine, surgery, pediatrics, obstetrics and gynecology, neurosurgery, medication, anesthesia, and blood transfusion cases. Students weighed these issues in small groups, using various discussion methods. After finishing the program, students completed a course evaluation questionnaire. RESULTS: The students rated this program as satisfactory, highly motivating, and helpful in preparing their future practices. They responded that although the cases were interesting, some were difficult. They stated that the small group discussion techniques encouraged them to take active part in the discussion and to consider the cases more deeply. CONCLUSION: Small group discussion of medical error cases is an effective method for students to study patient safety.

Evaluation of the Genotypes of the Lewis Blood Group in a Korean Population Using Direct Sequencing

BACKGROUND: The FUT2 and FUT3 genes determine the Lewis phenotype of red blood cells (RBCs). Recently, the Lewis genes, the secretor genes, and several mutations that cause Lewis negative and nonsecretor phenotypes have been identified. The purpose of this study was to analyze the gene frequency of FUT2 and FUT3 in a Korean population by comparing the use of the direct sequencing method to the use of the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method for mutation detection in the FUT2 and FUT3 genes. METHODS: RBCs and peripheral blood leukocytes were obtained from 225 apparently healthy volunteers to determine the phenotype and genotype of the FUT2 and FUT3 genes. Lewis phenotypes were determined on K3EDTA-stablized fresh blood samples using the column agglutination method. Lewis blood group genotyping was performed by use of the direct sequencing method. For the detection of T59G, C357T, and A385T mutations, the PCR-RFLP method was performed. RESULTS: The frequencies of the Lewis blood group phenotype were 12.4% for Le(a+b-), 70.7% for Le(a-b+), 11.1% for Le(a-b-) and 5.8% for Le(a+b+), respectively. Direct Sequencing of the FUT2 gene identified 92.2% C357T, 56.9% A385T, 0.4% G244A mutations and 1.8% del396. Direct Sequencing of the FUT3 gene identified 46.9% T59G, 30.4% G508A, 1.1% T202C, 1.1% C314T, 0.7% A1029G, 3.0% T1067A and 13.3% G1242A mutations. The PCR-RFLP method results were discordant in nine cases (1 case for C357T, 4 cases for A385T and 2 cases for T59G) as compared to the direct sequencing method results. CONCLUSION: We have determined the frequencies of FUT2 and FUT3 gene mutations in a Korean population. The use of the direct sequencing method was more accurate than the use of the PCR-RFLP method for the determination of the genotype of the FUT2 and FUT3 genes.

New Leukocyte Counting Method of Cerebrospinal Fluid: Using Transparent Ruler Tape / 대한진단검사의학회지

BACKGROUND: To enumerate leukocyte count in cerebrospinal fluid (CSF) is important for diagnosing bacterial meningitis. Using automated hematology analyzer for enumeration of leukocyte in CSF is below the sensitivity, so microscopic hemocytometric method is standard method. But this requires sufficient practical experience and has limitation of accuracy and stability. So we developed new manual method and evaluated it. METHODS: We designed new method using transparent ruler tape. We performed correlation, accuracy and precision test by counting leukocyte in diluted EDTA blood with three methods: new method, Neubauer and Nageotte hemocytometry. Twenty two CSF were used for stability test, which determines leukocyte count according to time (within one hour and after 2, 4 and 12 hr), by new method and Neubauer hemocytometry at room temperature. RESULTS: There was no clinical significant difference between three methods in correlation test, whereas Neubauer and Nageotte hemocytometry showed a bias to underestimation relative to the results obtained with new method in case with low leukocyte count. The new method showed the lowest CV and most accurate result. In stability test, leukocyte counts decreased being 44.4%, 72.1% of initial values after 2 hr, 14.8%, 31.1%, after 4 hr and 4.2%, 8.7%, after 12 hr, by Nageotte hemocytometry and new method, respectively. CONCLUSIONS: The new method we devised is simple, easy and applicable to use in a laboratory and offers advantages of improved precision and stability. It may be sufficient for replacing standard methods for leukocyte counting in CSF.

Development of a New Blood Typing Kit Using the Microfluidics Separation Technique

BACKGROUND: Blood typing is an essential test for transfusion. Generally, blood typing is performed using a slide test, tube test or microcolumn agglutination test. The aims of this study were to develop a new blood typing kit using micromachining, microfluidics and microseparation methods, and to evaluate the clinical usefulness of the new blood typing kit. METHODS: We designed and manufactured a blood typing microchip using polydimethylsiloxane (PDMS), which contained a microchannel (25~200 micrometer). The blood sample and antisera to be tested were dropped on the microwell for movement and mixing by capillary action. Once agglutination occurred, the microchannel acts as a filter and the blood type was determined by observation by the naked eye. To evaluate the newtyping kit, we tested sensitivity using artificially diluted blood and compared the results of the new typing method with the slide and tube methods using 70 samples. RESULTS: The new blood typing kit could differentiate a +4~+2 agglutination reaction, but could not detect a +1 agglutination reaction as observed by the naked eye. Among 70 samples, the results of ABO and Rh typing by the new typing method (n=66, > or = +2 agglutination reaction by the column agglutination method) were in accord with the results of the tube and slide methods, but couldnot detect agglutination in all 4 clinical samples, below a +1 agglutination reaction. CONCLUSION: The new blood typing kit is inadequate for routine use in the clinical laboratory due to low sensitivity, but with further improvement, it can be used economically, conveniently and objectively for blood typing without any special equipment. Moreover, the microfludics and separation method may be broadly applicable in other tests using the hemagglutination method.

Evaluation of a Blood Coagulation Factor XIII Assay Kit

BACKGROUND: Plasma coagulation factor XIII (FXIII) catalyzes the formation of covalent bounds between fibrin monomers, thus stabilizing the fibrin clot and increasing its resistance to fibrinolysis. Alteration of FXIII may contribute to bleeding, wound dehiscence and recurrent abortion. However, standard clotting tests cannot detect the FXIII deficiency. In this study, we evaluated a newly developed FXIII test kit (CoalinkTM, PeopleBio Inc., Seoul, Korea) in patients with various clinical conditions. METHODS: We evaluated the linearity and precision of the new FXIII test kit and compared the results of the new kit and the Pefakit FXIII assay. The FXIII was tested in idiopathic thrombocytopenic purpura (ITP) (n=40) patients, chronic renal failure (CRF) (n=20) patients, liver cirrhosis (LC) (n=40) patients, EDTA-induced pseudothrombocytopenia (EDTAIP) (n=10) patients, and in normal healthy persons (n=50). In the normal healthy persons, we determined a complete blood count (CBC), Ed-highlight-the second (n=50) is redundant. prothrombin time (PT) measurement and activated partial prothrombin time (aPTT) measurement and evaluated the results using the two assays. RESULTS: Serial dilution experiments with five samples provided good linearity (r2=0.9717). The intra- and inter assay precisions (CV) were 2.3~8.6% and 3.9~14.9%, respectively (n=20). There was a significant correlation between the use of the new kit and the Pefakit FXIII assay (r=0.8798, n=50). The FXIII activities of the normal healthy persons, ITP, CRF, LC and EDTAIP patients were 103.3+/-23.3%, 79.7+/-41.0%, 117.9+/-82.3%, 56.9+/-23.7% and 130.0+/-29.0%, respectively and they were significantly decreased in the ITP and LC patients (P<0.05). The rates below 80% of the FXIII level were 67.5% in the ITP patients, 90.0% in the LC patients, 35.0% in the CRF patients and 0.0% in the EDTAIP patients. FXIII activities were closely related to platelet count (r=0.832, P<0.05) and negatively correlated with PT (r=-0.389, P<0.05) and aPTT (r=-0.326, P<0.05). CONCLUSION: The new kit was determined to have good linearity and precision. Moreover, it was simple and rapid to perform. This method may prove useful for the evaluation of FXIII.

A case of fetal hydrops associated with maternal -D-/-D- phenotype / 대한산부인과학회지

Blood types are very important because they are associated with blood transfusion, the diagnosis of hematological diseases and the related diseases. Rh system is a major blood group system as ABO system. Major Rh antigen is Rh (D) antigen. Minor Rh antigens are Rh (C), (c), (E), (e) antigen. Most of people have Rh (C), (c), (E), (e) antigen but some people don't have these antigens. The hematologists call this blood type -D-/-D- blood phenotype. -D-/-D- phenotype is blood group that have (D) antigen without (C), (c), (E), (e) Antigen. This blood type is rare throughout the world. Especially, fetal hydops associated with anti-C,-c,-E,-e antibody is very rare. We experienced a case of fetal hydrops associated with anti-C,-c,-E,-e antibodies and report it with a brief of literatures.

The Seroprevalence of IgM and IgG Antibodies to Helicobacter pylori Among Maternal, Neonatal, and Cord Bloods in Korea / 임상검사와정도관리

BACKGROUND: Helicobacter pylori (H. pylori) infection has been known closely related with gastritis, duodenal ulcer and gastric cancer and is prevalent among Koreans. However, the infection route and the time are unclear, especially during perinatal period. The aim of this study is to investigate the relationship of H. pylori IgG and IgM antibody prevalences and titers between maternal, neonatal, and cord blood. METHODS: We collected 45 simultaneous maternal, neonatal, and cord bloods and 150 single cord bloods during delivery. The specific H. pylori IgG and IgM antibody levels were measured by enzyme-linked immunosorbent assay (ELISA) method. RESULTS: The H. pylori IgG antibody-positive rate for maternal, neonatal, and cord bloods were equal as 35.6% (16/45). The H. pylori IgG antibody levels of neonatal and cord bloods were 52.7% and 70.7% of maternal blood level. The H. pylori IgG antibody levels between maternal and cord bloods (r2 = 0.9725, p<0.05), maternal and neonatal bloods (r2 = 0.8569, p<0.05), and neonatal and cord bloods (r2 = 0.9437, p<0.05) were well correlated. Only one case of maternal blood was H. pylori IgM antibody positive and it's antibody level was 52.3 U/mL. CONCLUSIONS: In this study, we provided the sero-prevalence of H. pylori IgG and IgM antibodies and the relationship of antibody level of H. pylori IgG in maternal, neonatal and cord bloods. To elucidate the exact route and time of H. pylori infection, further studies including serial measurement of H. pylori IgG and IgM level in neonates will be needed.

The Reduction Effect of Leukocyte Fragments by Leukodepletion Filter / 대한수혈학회지

BACKGROUND: The efficiency of leukocyte removal filter is influenced by many factors. But, filtration efficiency of leukocyte fragments was not well known. We performed this study to evaluate whether the filtration efficiency for packed red blood cells can be influenced by leukocyte fragments according to storage time. METHODS: Leukocyte fragments in packed red blood cells (three units) which were artificially made by incubation for 4 hrs at 56degrees C and each four units of packed red blood cells according to storage time (0 days, 10 days, 20 days, and 30 days) were filtered using Sepacell R-500A (Asahi medical Co, Japan). The leukocyte concentrations of the pre-leukodepleted samples were estimated using an automated hematology analyzer (XE-2100, Sysmex, Japan). The ratio between the number of normal leukocytes and leukocyte fragments on Wright Giemsa stained slide was used in the analysis. The leukocyte concentrations of the post-leukodepleted samples were performed by the conventional counting methods using Nageotte hemocytometer. RESULTS: The ratios of fragmented to total leukocytes in packed red blood cells at pre- and post leukoreduction according to storage times were 1.5% and 16.3% within 1 days, 4.5% and 30.0% at 10 days, 6.3% and 35.0% at 30 days, and 8.3% and 42.5% at 40 days, respectively. Leukoreduction efficiencies of normal leukocytes in packed red blood cells were 99.99 +/- 0.01%, 99.97 +/- 0.02%, 99.98 +/- 0.01%, and 99.86 +/- 0.09%, respectively. The 36.0% of leukocytes in packed red blood cells were changed to fragmented leukocytes, residual fragmented leukocytes ratio was 95.0% and filter efficiencies of normal leukocytes was low(99.28%, p<0.05). CONCLUSIONS: The leukodepleted efficiency for leukocyte fragments were lower than for normal leukocytes. Leukocytes fragments may be influenced to lower the leukodepleted efficiency of normal leukocytes with storage time elapse.

Two Cases of ABO Discrepancy due to Hypogammaglobulinemia / 대한수혈학회지

Two cases of ABO discrepancy were observed in thirty-year old woman with gall bladder abscess and fifty-five-year old woman with hepatocellular carcinoma. Their red cells were typed as group O and their serum had only anti-A antibody. Absence of A and B antigens on their RBCs were confirmed by adsorption elution test and saliva test. The B transferase activities were not demonstrated in their serum. Their ABO genotypes were O/O by sequence specific polymerase chain reaction. Their serum protein electrophoresis showed hypogammaglobulinemia pattern, and immunoglobulin levels (IgG, IgA, IgM) were decreased (39 mg/dL, 46 mg/dL, <5 mg/dL and 63 mg/dL, 65 mg/dL, 12 mg/dL, respectively).

The genotyping of Kell, Duffy, and Kidd System in Korean / 대한수혈학회지

BACKGROUND: Among human blood group antigens, the genes for Kell, Duffy, and Kidd antigens have been recently identified, and those can play an important role in unexpected acute and delayed hemolytic transfusion reactions or hemolytic disease of newborns. The determination of blood group polymorphism at the genomic level facilitates the resolution of clinical problems that cannot be addressed by hemagglutination. They are useful to determine antigen types for which currently available antibodies are weakly reactive, type patients who have been recently transfused, identify fetuses at risk for hemolytic disease of the newborn and to increase the reliability of repositories of antigen negative RBCs for transfusion. METHODS: Two hundred peripheral blood samples were collected from normal population. Primer sets were used with slight modification from Reid M.E, et al. Bsm I, Ban I, and Mnl I were used from digestion of 5 uL PCR products. 10 uL of each digested-PCR products were electrophoresed on agarose or polyacrylamide gel with ethidium bromide staining. Kell, Duffy, and Kidd phenotypes (serologic types) were compared with respective genotypes by PCR-RFLP. RESULTS: The concordance rate was 100%: between genotype and phenotype 0 case(0%) K, 187 cases(100%) k; 22 cases(11.4%) Fy(a+b+), 171 cases(88.1%) Fy(a+b-), 1 case(0.5%) Fy(a-b+), 0 case(0%) Fy(a-b-); 95 cases(50.8%) Jk(a+b+), 39 cases(20.9%) Jk(a+b-), 53 cases(28.3%) Jk(a-b+), 0 case(0%) Jk(a-b-). In this study, Fyb frequency was 11.9% and it was equal to that of Japan and China. We analyzed each digested PCR product from 200 patients; Kell(187 cases), Duffy(194 cases), and Kidd(187 cases). CONCLUSIONS: The PCR-RFLP method can be effectively used for the Kell, Duffy, and Kidd typing and is particularly useful in cases where serological typing method is difficult as in autoimmune hemolytic anemia or recently transfused red blood cells in their circulation. Also, it is useful in cases of hemolytic disease in newborns and hemolytic transfusion reaction.

Evaluation of the RBC Lewis Blood Group Phenotypes and Genotypes of the FUT3 and FUT2 Gene / 대한진단검사의학회지

BACKGROUND: The Lewis and secretor gene determine the Lewis phenoytpe. Conventional Lewis blood grouping is difficult because of the presence of nongenuine Lewis negative individuals. Recently, the Lewis gene (FUT3), the secretor gene (FUT2), and several mutations that cause the Lewis negative and the nonsecretor phenotypes were identified. The purpose of this study was to compare Lewis phenotypes determined by commercially available three pairs of monoclonal antibodies with the Lewis and secretor genotypes. METHODS: RBCs for phenotyping and peripheral blood leukocytes for genotyping of FUT3 and FUT2 gene were obtained from 184 apparently healthy volunteers. Lewis phenotypes were determined on K3EDTA-stablized fresh blood samples using three pairs of commercially available monoclonal antibodies, one of which was the column agglutination method and the others were the tube agglutination methods. Lewis blood group genotyping was performed by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method to detect T59G, G428A, C357T, and A385T mutations. RESULTS: The frequencies of the Lewis blood group phenotype were Le(a+b-) 15.0%, Le(a-b+) 65.8%, Le(a-b-) 14.8%, and Le(a+b+) 4.3%, respectively. The Lewis blood group phenotypes determined by three pairs of monoclonal antibodies were 93.5%, 93.5% and 89.1% in accordance with the genotypes. The frequencies of Le, le, Se and se alleles were 64.4%, 35.6%, 48.6%, and 51.4% and we have newly detected 4 cases with only one A385T mutation. All of the Le(a+b+) phenotype cases have both C357T, and A385T homozygotic mutations. CONCLUSIONS: The PCR method may be effectively used for the genotyping of the FUT3 and FUT2 genes and offers an attractive alternative to Lewis phenotyping using hemagglutination method.