Analysis for Human Immunodeficiency Virus 1 Subtype in Korean Blood Donors / 대한수혈학회지

BACKGROUND: Genetic variants of virus appear to differ depending on the country, race, infection route, and so on. To characterize the main HIV subtype in infected blood donors and inquire about the route of HIV infection, we analyzed HIV subtype for samples that showed reactive results on the anti-HIV 1/2 and HIV-1 NAT test from September 2007 to February 2010. METHODS: To identify the HIV-1 subtype of the 90 samples that showed reactive results on the anti-HIV test and HIV-1 NAT, we performed HIV 1/2 Western blot assay, HIV RNA quantitative assay, HIV-1 nested PCR, and HIV-1 RNA sequencing. RESULTS: A total of 85 samples (94.4%) were confirmed to be HIV-1 subtypes. Among them, 82 samples (96.5%) were subtype B; and subtype A, C, and G was confirmed for one case each (1.2% for each case). We could not identify the subtype of the other five samples. One of them was amplified by nested PCR, but was not confirmed of the subtype, and four samples were not amplified even by nested PCR. CONCLUSION: The main HIV-1 subtype among the HIV-infected blood donors was confirmed to be subtype B. In addition, we identified one case each of HIV-1 subtype A, C, and G, which was not detected in blood donors in the past. It appeared that the route of HIV infection in Korea had become complicated. Therefore, we concluded that continuous research for HIV subtype analysis and efficient management of blood donors is needed.

Development and Preliminary Evaluation of a Leukocyte Removal Aptamer Filter / 대한수혈학회지

BACKGROUND: Leukocyte reduction filters are widely used to prevent transfusion reactions caused by leukocytes in blood components. Commercial filters are not sufficient for removal of leukocytes for prevention of transfusion associated graft-versus-host disease; therefore, irradiation of blood components was performed using expensive equipment. Techniques using an aptamer substituted for antibody have been developed and are available in clinical areas. The purpose of this study is to develop the aptamer filter system and to evaluate its efficiency and the possibility of its clinical application. METHODS: Aptamers targeted to CD45 were selected by the Postech Aptamer Initiative. The aptamer filter in which aptamers attached to beads were bound to leukocytes and removed by magnetic field was developed. Filtration of 14 units of leukoreduction-red blood provided by Korean Red Cross Blood Services was performed using aptamer filters. Leukocyte removal rate and red cell recovery rate were evaluated and bacterial culture was performed. RESULTS: After filtration using the aptamer filters, 45.6% of leukocytes were additionally removed and the red cell recovery rate was 92.8%. No growth in the bacterial culture was observed. CONCLUSION: In order to apply the cell depletion technique utilizing an aptamer to blood filter system, we developed and evaluated the aptamer filter system. Through improvement of the binding efficiency of the aptamer and the filtering process, and application of the various aptamers for other different cells, we suggest that this technique can be applied in the clinical area, such as a substitution for the irradiation process for TAGVHD prevention.

Analysis of the Coagulation Factors of Donated Plasma for Effective Utilization / 대한수혈학회지

BACKGROUND: The aim of present study was to assess the effect of different freezing time after phlebotomy on the activity of coagulation factors in frozen plasma and to evaluate which source plasma for clotting factor fractionation is appropriate for use. METHODS: Blood plasma units rejected due to a high level of ALT were divided into four groups depending on freezing time after phlebotomy, and each unit of the four groups was assayed for six different clotting factors and blood type. SAS 9.2 was used for statistical analysis of data. RESULTS: A decrease was observed in the activities of FVIII of the plasmas, in the following order: PL-A>FFP>FP(8-24)approximatelyFP(24-72). Results of the assay also showed that the levels of FVIII were significantly higher in the AB type plasmas than in the O type plasmas. PL-A and FFP units met the current quality requirements of the Korean Red Cross, in which the FVIII activity should have more than 0.7 IU/mL in more than 75% of the source plasma, as 85.0% and 82.5%, respectively. On the other hand, FP24 met the Canadian (Quebec) requirements for the source plasma, in which the FVIII activity should have more than 0.52 IU/mL in more than 75% of the source plasma, as 82.6%. CONCLUSION: For use of plasma frozen within 24 hours after phlebotomy (FP24) and plasma of specific blood type, European Pharmacopeia and WHO guidelines on quality control should be adopted for production of plasma-derived coagulation factors in Korea.

Establishment of Plasma Working Standards for the Performance and Quality Assurance of NAT Screening Tests for HIV, HCV and HBV / 대한수혈학회지

BACKGROUND: Since Jan. 2012, for performance evaluation of viral reagents, analysis of domestic samples has been recommended in order to obtain approval from the KFDA when they are first introduced to Korea. This regulation requires the standard domestic materials driven from locally infected samples. We tried manufacturing the plasma working standards of HBV, HCV, and HIV NAT using a mixed titer of viral loads. METHODS: Forty three HBV DNA positive plasmas, 25 HCV RNA positive plasmas, and 26 HIV RNA positive plasmas were evaluated according to viral load and genotype. Several plasma units, which had high-titer viral loads and the common viral genotypes in Korea, were selected as the source materials for each viral standard. To adjust the appropriate concentration based on the detectable range of variable viral reagents, the source plasma was diluted to several concentrations, divided into small vials, and analyzed for quantification. RESULTS: The 13 plasma working standards, which had variable viral loads for the mixed titer performance panel of HIV, HCV, and HBV NAT, were produced. CONCLUSION: These national standard materials were first produced in order to supply the mixed titer performance panel for the viral NAT reagent of the level IV transfusion related high-risk group in Korea.

Chromosomal Abnormalities in Myelodysplastic Syndrome with Near-tetraploidy: A Case Report

Massive hyperdiploidy and tetraploidy are rare cytogenetic abnormalities in myelocytic malignancies, especially in myelodysplastic syndrome (MDS). These abnormalities are known to be associated with leukemogenesis, leukemic transformation and poor prognosis. We report here the first case of MDS with near-tetraploid cytogenetic abnormality in Korea. A 80-yr-old male was diagnosed with refractory anemia with excess blasts-2 (RAEB-2). Bone marrow aspiration smear showed 16% of blasts, which were large sized myeloid blasts with irregular margins and cytoplasmic vacuolation. Cytogenetic analysis of bone marrow cells revealed numercal and structural cytogenetic abnormalities including near-tetraploidy in 8 of 20 metaphases: 45,XY,add(1)(p36.1),del(10)(p11.2),del(11)(q13),-12,-16,der(17)t(11;17) (q13;q21),add(20)(q13.1),+mar[8]/85~90,idemx2[cp8]/46,XY[4]. After chemotherapy with decitabine, he showed pancytopenia during follow-up period and died of sepsis 14 months after the diagnosis.

Evaluation of the Automated Instrument QWALYS-3 for Unexpected Antibody Screening / 대한수혈학회지

BACKGROUND: The use of automated techniques reduces the impact of human errors in blood banking and it improves the standardization and the quality of the achieved results. Erythrocyte Magnetized Technology (EMT) is now being widely used due to its simplicity and efficiency for detecting alloantibody. We evaluated the antibody screening test of the QWALYS-3 (DIAGAST, Loos Cedex, France). METHODS: The evaluation focused on antibody screening using the QWALYS-3 as compared to the standard manual tube method and the Ortho BioVue system in clinical samples (n=100) and frozen stored samples (n=64), which had RBC alloantibody. RESULTS: Using the manual tube method, the sensitivity of antibody screening was 100% by the QWALYS-3 and 42.8% by the Ortho BioVue in the clinical samples (n=7) and 2 results were discrepant by the QWALYS-3 for negative samples. For the known antibodies from the frozen stored samples (n=64) this correspondence rate amounted to 93.7% (n=60). CONCLUSION: The QWALYS-3 system displayed a good match rate with the Ortho BioVue system (92%). It also showed reliable results for the general accuracy when compared to the manual method (concordance rate: 98%). The QWALYS-3 system will facilitate the automation of routine antibody screening with high reliability, sensitivity and specificity compared to the standard manual methods.

A Case of Hemophagocytic Lymphohistiocytosis with Clonal Karyotype Abnormalities

There have been a few reports of hemophagocytic lymphohistiocytosis (HLH) with chromosomal abnormalities. Clonal chromosomal abnormalities in HLH patients are usually found in association with hematologic malignancies and rarely with epstein-barr virus (EBV) infection. Here, we report a fatal case of HLH with clonal karyotype abnormalities. A 75-yr-old man was admitted with persistent anorexia and high fever. Laboratory data revealed pancytopenia, hypofibrinogenemia, hyperferritinemia, prolonged prothrombin time and activated partial thromboplastin time, and marked elevated level of serum transaminases. In real time-PCR using whole blood, EBV DNA was not detected but cytomegalovirus (CMV) DNA was detected. The bone marrow aspiration smear showed hyperplasia of mature histiocytes with prominent hemophagocytosis. In chromosomal analysis of bone marrow aspirates, complex chromosomal abnormalities were found. In spite of steroid pulse therapy and antibiotic treatment, he died of disseminated intravascular coagulopathy.

ABO Genotyping of a Neonate with Mixed Field Agglutination / 대한수혈학회지

Mixed field agglutination is an important, but rare phenomenon of ABO blood grouping. Contrary to adults, neonatal red blood cells are immature and they present a weak ABO expression, and sometimes this result in a mixed field agglutination pattern. We report here on a case of a neonate who presented with mixed field agglutination on the ABO blood grouping during serologic testing and the neonate had a normal ABO genotype.

Determining the Cause of Discrepant Results in HCV NAT Comparative Tests / 대한수혈학회지

BACKGROUND: The Korean Red Cross blood laboratory centers have been performing comparative tests for NAT reactive specimens since February 2005. However, five discrepant specimens were found in HCV-diluted specimens between 2007 and 2008 and the reasons for this has been investigated. METHODS: For the five discrepant specimens, the HCV RNA concentration was measured in 5 tubes for each speciment. Subsequently, in order to compare the sensitivity of the low titer specimens measured by RT-PCR and TMA, comparative tests of diluted samples were examined six times per sample. Finally, the genotype was tested in order to determine the characteristics of the discrepant samples. RESULTS: Result of the quantitative tests for HCV RNA demonstrated that aliquots of the plasma bag were homogenous in term of viral load quantitation. As a result of the comparative test, all samples were found to contain over 1.0x10(1) IU/mL as detected by the two analytical systems. In contrast, those less than 1.0x10(1) IU/mL were not entirely detected by the two systems. CONCLUSION: It was impossible to completely detect using the two NAT system and the detection rates for both systems were equivalent for the samples examined. In particular, with respect to HCV, it may be undetectable on the NAT test because viral load decreases rapidly before and after sero-conversion. This result indicates that anti-HCV and NAT should be performed together as an HCV screening test prior to blood donation.

A Case of Mild Hemolytic Disease of the Fetus and Newborn - The First Case of Anti-Di(b) Identified on Prescreening Test during Pregnancy - / 대한수혈학회지

The Di(a+b-) phenotype is extremely rare among Caucacians and mostly confined to mongoloids. The clinical significance of anti-Di(b) is incompletely recognized. The authors report here a mild case of hemolytic disease of the fetus and newborn (HDFN) due to anti-Di(b). The mother was Di(a+b-) with anti-Di(b), which was detected by antenatal screening. She delivered a Di(a+b+) infant with a positive direct antiglobulin test and mild hemolytic disease. The infant was treated by phototherapy and subsequently recovered without the need for exchange transfusion. We suggest that the screening program as routine prenatal care is necessary.

Characteristics of Intron 6 Nucleotide Sequences of cis-AB in Koreans / 대한수혈학회지

BACKGROUND: The cis-AB is a very rare phenotype in the ABO blood group system. It corresponds to a special ABO allele that encodes glycosyltransferase that is capable of synthesizing both A and B antigens. Until now, the exon 6 and 7 gene sequences of cis-AB alleles are well known. In this study, we report on the intron 6 sequence structure of the cis-AB allele. METHODS: Standard serologic tests for the ABO blood group phenotypes were performed in four cis-AB samples. Allele-separation by cloning and subsequent sequencing was carried out. RESULTS: The results showed that intron 6 of cis-AB is almost identical to the A101 allele except for three single nucleotide polymorphisms at nucleotide positions 163, 179 and 662, where the nucleotides of the A101 replace those of B101. CONCLUSION: The intron 6 sequences of cis-AB in Koreans have both A101 and B101 blood group sequences.

A Study on the Improvement of Blood Donor Health Questionnaire / 대한수혈학회지

BACKGROUND: For the safety of blood resources, the Blood Donor Health Questionnaire (DHQ) should be modified and improved allowing donors to answer questions with further accuracy. To accomplish this, it is essential to identify any part of this questionnaire that is donors find inconvenient. METHODS: The problems of the current DHQ were examined through a poll of donors at the Korean Red Cross and other hospital blood service centers from November 2008. We also compared the structure and contents of the Korean DHQ to similar document in eight other countries. RESULTS: Donors thought that the current DHQ was too complicated, took too much time (27.3%) and probed too much into a donors private life (51.2%), making it difficult to answer honestly. The Korean DHQ focuses on a deferral period and uses special medical terminology in order for an interviewer to make easy decisions regarding donor eligibility. In contrast, other questionnaires tend to focus on a donor's recall of memory, use simple vocabulary, and emphasize donor's duties, and therefore, these documents are easy for donors to understand and complete CONCLUSION: Donor-oriented DHQs using simply terminology, help donors with memory recall and emphasize a donor's duty. Also, such a document allows donors to answer frankly. Therefore donor-oriented DHQs provide a great degree of blood resource safety than interviewer-oriented DHQs.

Ultrastructural Changes of Nerve Fibers Using a Neuropathic Pain Model in a Rat / 대한마취과학회지

BACKGROUND: Neuropathic pain is part of the symptom complex known as peripheral neuropathy. Sensory loss, muscle weakness, atrophy, and decreased tendon reflexes are more common than pain in neuropathic disease. Recently, Bennett and Xie reported that when the sciatic nerve of a rat is loosely ligated, the rat develops pain syndrome similar to that observed in neuropathic pain states in a human. Anatomical and physiological studies to date indicate that the major pathological finding in large diameter myelinated fibers distal to the ligatures was a complex loss of response while in small myelinated fibers there were was only subtle changes. However, a more extensive analysis of the various nerve fiber groups in the damaged sciatic nerve is required for a better understanding of the pathophysiology of the present neuropathy. METHODS: To evaluate the damage and regeneration of all caliber of peripheral nerve, we performed an electron microscopic analysis of the sciatic nerve after four loose ligatures were applied. Cross- sectional photomicrographies of regions distal to the ligatures were studied. A peripheral mononeuropathy was produced in adult rats by tying 4 ligatures loosely around the common sciatic nerve. The distal part of the ligated common sciatic nerve was severed in 2 rats of each group at 1 day, 3 days, 1 week, 2 weeks and 4 weeks respectively. The severed nerves were prepared for electron microscopic examination and pathologic changes were observed under the electron microscope. RESULTS: The ultrastructural changes after ligature application were as follows: At 1 day, the axon of A-beta fiber was shrunken and detached from the myelin sheath. C-fibers were mildly edematous and A-delta fibers appeared to be normal. On the 3rd day, the axoplasm of A-beta fibers was more shrunken, containing swelling of microorganelles and irregularly thickened myelin sheath. C-fiber showed some degrees of degeneration. A-delta fibers revealed mild degeneration and interstitial edema was also noted. At 1 week, the myelin sheaths of A-beta fibers were severely irregular in appearance with marked axonal loss. Many myelin fragments were phagocytosed in the cytoplasm of adjacent Schwann cells. At 2 weeks, A-beta fibers predominantly disappeared and many fragmented myelin sheaths were ingested in the Schwann cell. In some areas, A-beta fibers partially regenerated, which involved remyelination and an increase in the numbers of microorganelles of the Schwann cells. C-fibers were also regenerated. At 4 weeks after sciatic nerve ligation, A-beta fibers regenerated and myelin ovoids were noted within the axoplasm of the A-beta fibers. Myelin ovoids were found in the Schwann cell cytoplasm. A-delta fibers and C-fibers appeared ultrastructurally well-regenerated and had a relatively normal distribution. CONCLUSIONS: We found that maximal nerve degeneration was observed at 2 weeks after sciatic nerve ligation, thereafter, nerve regeneration was noted at 4 weeks after sciatic nerve ligation.

A Study of the Need of Clinical Expert System for Acid-Base Diagnosis / 대한임상병리학회지

BACKGROUND: Rapid and accurate diagnosis of multiple acid-base disorders is not so easy. The authors evaluated a need of a computer-aided diagnosis program for acid-base disorders through the questions of 25 cases with acid-base disorders in 105 physicians. METHODS: For 39 college graduated students and 66 housestaffs, we asked them to diagnose 25 sample cases of acid-base disorders, which were selected from the published 36 cases (Horn et. al. 1992, Halperin and Goldstein 1994, Walmsley and White 1988). The 25 questions consisted of 15 cases with single disorder, 8 cases with double disorder and 2 cases with triple disorder. RESULTS: The 39 college graduated students diagnosed correctly in 55.4%; 79.6% in single disorders, 23.7% in double disorders and 0.0% in triple disorders. The 36 housestaffs (answer rate: 55.4%) diagnosed correctly in 61.7%; 82.4% in single disorders, 38.2% in double disorders and 0.0% in triple disorders. CONCLUSIONS: The data suggest that the physicians can't diagnose acid-base disorders well and that they needs a computer-aided expert system to diagnose the acid-base disorders and to ensure uniform quality of patient care.

The Changes of Plasma Concentration of Lidocaine Associated with Epidural Anesthesia / 대한마취과학회지

Epidural anesthesia is widely practiced for lower aMominal surgery and painless delivery in many hospitals and its complications are much lower than that of spinal anesthesia, Lidocaine is a most commonly using local anesthetic in clinics and it has been used exten-sively for epidural anesthesia with very good results. It has a rapid onset of acton with a good analgesic effect and a reasonable duration of action (about l - 11/2 hr). The primary purpose of this study is to observe toxic levels of plasma lidocaine concentration in patients who receive different doses of lidocaine epidurally. Twenty-two adult patients were divided into three groups. With patients in a sittng position, single epidural anesthesias were perfomed with a 17 gauge Tuohy needle. The 2% lidocaine mixed with 1: 200,000 epinephrine was used as a local anes- thetic. Of these, 7 patients in group l were given 400 mg and 8 patients in group 2 were given 500 mg and 7 patients in group 3 were given 600 mg. The plasma concentrations of lidocaine were measured by immunofluorescence assay at the intervals of 5, 10, 15, 30, 45 and 60 minutes. The peak plasma concentrations in the group 1, 2 and 3 were 2.24+/-0.51, 3.84+/-0.90 and 4.36+/-1. 16ug/ml (mean+/-SD) respectively. The peak plasma concentrations were measured 15 and 30 minutes after the epidural injection in group 1, 2 and 3 respectively. The maximum peak plasma concentration among 22 patients was 6.48ug/ml. These results indicate that the injection of 2% lidoeaine 500, 600mg with epinephrine is considered dangerous to some extent because the plasma concentrations of lidocaine exceed toxic levels occasionally.

Paraplegia Following Inadvertent Epidural Administration of Potassium Chloride / 대한마취과학회지

A 25-year-old female came in for an explo-laparotomy. The operation was performed under epidural anesthesia. Potassium chloride (40 mEq20 ml) instead of 2% lidocaine mixed with 1:200,000 epinephrine was accidently injected via a Tuohy needle into the epidural space. The patient showed an unexpected prolonged motor and sensory paralysis below the T, cord level for about 8 hours after the epidural injection. The patient recovered without a permanent neurologic sequale after 8 hours. However, the patient developed pulmonary edema resulting from rapid fluid therapy for unusual hypotension which developed 12 hours after the potassium chloride injection. The pulmonary edema was treated by oxygen therapy using a nonrebreathing oxygen mask, a dopamine drip and an intravenous injection of lasix 20 mg. The patient completely recovered from the pulmonary edema on the 3rd postoperative day and went home with no complaints.

Tonic - Clonic Convulsions of the Lower Limbs following Spinal Anesthesia Using Meperidine / 대한마취과학회지

Recently we encountered consecutive but transient tonic-clonic convulsions and motor weak-ness in the lower limbs of a patient during the recovery from meperidine spinal aneathesia for Cesarean section. The patient revealed no previous history of neurologic disease, and there was no suspicious cause to explain the phenomenon. The episodes of convulsion were suppressed with intravenous diazepam. The mechanisms are vaguely suggested that the spinal inhibitory neurons are inhibited by the local anesthetic effects of spinal meperidine and an effect similar to the convulsive action of intraspinal morphine occurs.

A case of promyelocytic crisis of chronic myelogenous leukemia / 대한임상병리학회지

No abstract available.

Bicanalicular Silicone Intubation of Canalicular Laceration

Thirty cases of lacerated canaliculi were repaired by bicanalicular silicone intubation(Pilling(R) Intubation Set)with use of a silicone sponge for nasal fixation. The results were as follows: Canalicular laceration occurred mainly in males(25 cases), and in young age group(10-39yrs). An isolated inferior canalicular laceration was the most common and occurred in 23 cases. There was no significant difference in laterality between right and left. The average follow up period was 13.8 months and final results were better in patients who were repaired earlier and had a longer intubation time. Postoperative complications occurred in 7 cases and all of them were canalicular slits. Canalicular slits were not related to the duration of intubation and did not affect the final results significantly. The advantages of this operation were good cosmetic results, minimal irritation by tube, and easy removal of tube.