• The Journal of Korean Society of Virology
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• v.28 no.1
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• pp.31-38
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• 1998

In Korea, all domestic made test systems for detecting antibodies in HIV-1 contain the antigens from human immunodeficiency type 1 (HIV-1) subtype B. However, because HIV-1 subtype O is significantly different in amino acid sequences from all other subtypes of HIV-1, there has been a need for developing a test for detecting antibodies in subtype O. For this purpose, the entire nucleotide sequence corresponding to the extracellular domain of the transmembrane glycoprotein of HIV-1 subtype O was synthesized with consideration of Escherichia coli condon usage. Various regions of the extracellular domain were cloned into E. coli expression vectors and tested for levels of protein production. The nucleotide sequence, named ECTM, that can encode a 129 amino acid-long peptide, was found to be expressed at a high level in E. coli. The protein of approximately 17 kDa specifically reacted with sera from individuals infected with HIV-1 subtype O. The ECTM protein was purified to near homogeneity by the CM-T gel chromatography, using concentrated, denatured inclusion bodies. In Western blot analysis, the purified viral antigen reacted with sera from individuals infected with subtype O more efficiently than subtype B. The enzyme linked immunoabsorbent assay (ELISA) system was developed using the subtype O viral protein and compared with the commercially available kit lacking the antigens from subtype O. The ELISA kit containing the subtype O antigen ECTM alone efficiently reacted with sera from individuals infected with subtype O. The subtype O antigen-containing kit produced a positive absorbence even when sera were diluted 512-fold, suggesting a high sensitivity. The commercially available kit also reacted with subtype O sera, but produced a negative result at a dilution of 8-fold. Our results suggest that the currently available kit may not be able to efficiently detect subtype O sera and that the viral protein developed in this study may be added to the current system to maximize the detection of sera from individuals infected with subtype O.

• Pediatric Infection and Vaccine
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• v.24 no.1
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• pp.23-30
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• 2017

Purpose: The objective of this study was to compare the clinical characteristics of influenza A and B infections and analyze the effect of oseltamivir in hospitalized children. Methods: We investigated children under the age of 15, who were diagnosed with influenza A/H1N1, A/H3N2, or B from January to April 2014. The subjects were admitted to the Changwon Fatima Hospital and diagnosed using a rapid antigen test from nasopharyngeal swabs. The medical records of the patients were retrospectively reviewed. Results: A total of 302 pediatric patients with influenza were enrolled. Influenza B infection was the most common type (n=187, 61.9%), followed by A/H3N2 (n=100, 33.1%) and A/H1N1 (n=15, 5.0%). Compared to patients diagnosed with influenza A, patients diagnosed with influenza B were older (P=0.005), and the duration of fever was significantly longer (P=0.001). A total of 161 patients (53.3%) had been vaccinated against influenza during the season, before admission. Among the patients infected with A/H3N2 and B, the duration of fever was shorter in oseltamivir recipients compared to oseltamivir non-recipients (P=0.026 and P=0.004, respectively). Conclusions: There were significant differences between influenza A and B groups in terms of age, demographics, and clinical course. Although the effectiveness of oseltamivir on influenza differs according to the type of influenza, our data provides evidence that oseltamivir is beneficial for both A and B infections.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.31 no.3
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• pp.199-218
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• 2005

Purpose of Study: Peripheral nerve regeneration depends on neurotrophism of distal nerve stump, recovery potential of neuron, supporting cell like Schwann cell and neurotrophic factors such as BDNF. Peripheral nerve regeneration can be enhanced by the conduit which connects the both sides of transected nerve. The conduit maintains the effects of neurotrophism and BDNF produced by Schwann cells which can be made by gene therapy. In this study, we tried to enhance the peripheral nerve regeneration by using calcium phosphate coated porous conduit and BDNF-Adenovirus infected Schwann cells in sciatic nerve of rats. Materials and Methods: Microporous filter which permits the tissue fluid essential for nerve regeneration and does not permit infiltration of fibroblasts, was made into 2mm diameter and 17mm length conduit. Then it was coated with calcium phosphate to improve the Schwann cell adhesion and survival. The coated filter was evaluated by SEM examination and MTT assay. For effective allogenic Schwann cell culture, dorsal root ganglia of 1-day old rat were extracted and treated with enzyme and antimitotic Ara-C. Human BDNF cDNA was obtained from cDNA library and amplified using PCR. BDNF gene was inserted into adenovirus shuttle vector pAACCMVpARS in which E1 was deleted. We infected the BDNF-Ad into 293 human mammary kidney cell-line and obtained the virus plaque 2 days later. RT-PCR was performed to evaluate the secretion of BDNF in infected Schwann cells. To determine the most optimal m.o.i of BDNF-Ad, we infected the Schwann cells with LacZ adenovirus in 1, 20, 50, 75, 100, 250 m.o.i for 2 hours and stained with ${\beta}$-galactosidase. Rats(n=24) weighing around 300g were used. Total 14mm sciatic nerve defect was made and connected with calcium phosphate coated conduits. Schwann cells$(1{\times}10^6)$ or BDNF-Ad infected Schwann cells$(1{\times}10^6)$ were injected in conduit and only media(MEM) was injected in control group. Twelve weeks after surgery, degree of nerve regeneration was evaluated with gait analysis, electrophysiologic measurements and histomorphometric analysis. Results: 1. Microporous Millipore filter was effective conduit which permitted the adhesion of Schwann cells and inhibited the adhesion of fibroblast. We could enhance the Schwann cell adhesion and survival by coating Millipore filter with calcium phosphate. 2. Schwann cell culture technique using repeated treatment of Ara-C and GDNF was established. The mean number of Schwann cells obtained 1 and 2 weeks after the culture were $1.54{\pm}4.0{\times}10^6$ and $9.66{\pm}9.6{\times}10^6$. 3. The mRNA of BDNF in BDNF-Ad infected Schwann cells was detected using RT-PCR. In Schwann cell $0.69\;{\mu}g/{\mu}l$ of DNA was detected and in BDNF-Adenovirus transfected Schwann cell $0.795\;{\mu}g/{\mu}l$ of DNA was detected. The most effective infection concentration was determined by LacZ Adenovirus and 75 m.o.i was found the most optimal. Conclusion: BDNF-Ad transfected Schwann cells successfully regenerated the 14mm nerve gap which was connected with calcium phosphate coated Millipore filter. The BDNF-Ad group showed better results compared with Schwann cells only group and control group in aspect to sciatic function index, electrophysiologic measurements and histomorphometric analysis.

• Childhood Kidney Diseases
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• v.7 no.2
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• pp.133-141
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• 2003

Purpose : Idiopathic Membranous Nephropathy(IMN) is a rare renal disease in children. To help better understanding of its clinical course and treatment strategies, we reviewed the clinical manifestations and pathological findings of children with IMN. Methods : Among 58 cases with MN, from 1977 to 2003, 42(72.4%) were hepatitis B virus (HBV) associated and 16(27.6%), 6 males and 10 females, were idiopathic. All cases diagnosed aster 2000 were IMN. Several clinicopathological findings(sex, onset age, proteinuria, serum albumin, cholesterol, creatinine clearance, tubulointerstitial changes, glomerular sclerosis, hypertension, renal vein thrombosis, the use of ACE inhibitor, and immunosuppressive therapy) were compared between the remission and the non-remission group of the patients with IMN. Results : The median onset age was 13.4 years. Clinical manifestations were nephrotic syn-drome(7 cases, 43.8%), gross hematuria(5 cases, 31.3%) and microscopic hematuria with proteinuria(3 cases, 18.8%). Hypertension, hypocalcemic tetany and renal vein thrombosis were accompanied in 2, 1 and 2 cases, respectively. In addition to the typical findings of MN, the kidney biopsies showed segmental sclerosis(5 cases, 31.3%) or global sclerosis(6 cases, 37.5 %), diffuse crescents(1 case), and mild(11 cases, 68.7%) or moderate tubulointerstitial changes(3 cases, 18.8%). Thirteen cases(86.7%) received oral steroid. Among them 2 cases received cyclophophamide and 1 received cyclosporin as well. Ten cases(62.5%) received ACE inhibitors. In the patients followed up, 7 cases(46.7%) became free from proteinuria (remission group) while 8(53.3%) presented continous proteinuria (non-remission group), two (13.3%) of which progressed to renal failure. Clinicopathological findings showed no significant differences between the two groups. Conclusion : With HBV vaccination, HBV associated MN decreased markedly and IMN has taken up most of MN in children. For better understanding of this rare disease, a prospective multicenter study of the clinical course and treatment strategies should be done.

• Horticultural Science & Technology
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• v.33 no.2
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• pp.210-218
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• 2015

Melon leaves showing yellowing symptoms were analyzed using electron microscopy and RT-PCR for major cucurbit-infecting-viruses (CMV, MNSV, CGMMV, SqMV, WMV, KGMMV, PRSV and ZYMV) reported in Korea, but these viruses were not detected. As the result of further analysis by next-generation sequencing (NGS), the virus was identified as Cucurbit aphid-borne yellows virus (CABYV), and then confirmed by RT-PCR using CABYV-specific primers. When photosynthetic capacity was measured based on chlorophyll fluorescence yield (ChlFY), the leaves of the diseased plants showed $4.09{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$, which was one-third of the readings observed for unaffected normal plants ($12.36{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$). The root functions of plants affected by leaf yellowing symptoms (LYS) was $0.28mg{\cdot}g^{-1}$, about half that measured for the normal unaffected plants ($0.48mg{\cdot}g^{-1}$). Cytological observations revealed that there were no morphological differences in the palisade parenchyma and mesophyll spongy cells of the leaves between the diseased and the normal plants. However, the same leaf cells of the affected plants contained more starch granules compared to those of the normal, unaffected plants. We conclude that the LYS of muskmelon is not merely a physiological disorder but a viral disease caused by CABYV and spread by aphids.

• Pediatric Infection and Vaccine
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• v.22 no.2
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• pp.55-62
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• 2015

Purpose: Varicella Zoster Immune Globulin (VZIG) is available in Korea for post-exposure prophylaxis of the Varicella-zoster virus (VZV) in high-risk patients. In July 2013, the United States Centers for Disease Control and Prevention (US CDC) recommended extending the time for administration of VariZIG$^{(R)}$ from within 96 hours up to 10 days after VZV exposure. This study was performed to analyze the effectiveness of VZIG prophylaxis between the two groups of patients who received VZIG within 96 hours and more than 96 hours of exposure to varicella. Methods: A retrospective chart review was performed in pediatric patients who received VZIG at Samsung Medical Center, Seoul, Korea from January 2001 to December 2012. Results: A total of 91 patients were identified. Fifty-seven patients were male (62.6%) and the median age was 5.91 years. Thirty-nine patients (42.9%) were exposed to VZV in the hospital. Underlying diseases were solid tumors (41.8%), hematologic malignancies (40.7%), and others (17.5%). Forty-five patients (49.5%) were hematopoietic cell transplant recipients. Seventy-four patients (81.3%) received VZIG within 96 hours after VZV exposure. There was no significant difference in the development of chickenpox between the two groups (2.7% vs. 5.9%, P=0.4664). In 22 seronegative patients, we also observed no significant difference between the groups in terms of the development of chickenpox (6.6% vs. 0%, P=0.667). Conclusions: This study showed that the effectiveness of VZIG for the prevention of chickenpox was comparable between patients who received VZIG within 96 hours and those who received VZIG more than 96 hours after exposure to VZV.

• Pediatric Infection and Vaccine
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• v.5 no.2
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• pp.258-266
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• 1998

Purpose : Cases of adenoviral penumonia with rapidly progressive clinical course were experienced. We reviewed these patients in viewpoint of clinical manifestation and adenoviral serotypes. Methods : Culture and indirect immunofluorescence for respiratory viruses including respiratory syncytial virus, influenza virus, parainfluenza virus, adenovirus was done with nasopharyngeal aspirates from patients who admitted due to respiratory infections in Fatima Hospital, Masan from Nov. 1996 to Jul. 1997. Cultured adenovirus was serotyped by both neutralization and hemagglutination inhibition test. Medical records were reviewed for 5 patients with respiratory failure from adenovirus was isolated and serotyped. Results : The total number of examined patients was 460 patients. We isolated respiratory viruses in 143(30.9%) patients. Adenovirus was isolated from 66 out of 143(46.2%) patients. During Jan 1997 to May 1997, five patients with ages of 18 days to 11 months who were infected by adenovirus and had high fever with dyspnea and required assisted mechanical ventilation. One patients discharged against doctor's advice then died. Two of four patients had complications of disseminated intravascular coagulation; two had bronchiolitis obliterans. Two isolates were serotype 7, and one was serotype 5, and two were untyped. Conclusion : Severe pneumonia caused by serotype 7 continued to occur in 1997 following the epidemic in 1996, and severe pneumonia may also be caused by serotype 5 and other serotypes.

• Journal of Veterinary Clinics
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• v.29 no.4
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• pp.309-314
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• 2012

The study was undertaken to evaluate sensitivity and specificity of rapid Avian Influenza (AI) and Newcastle Disease virus (NDV) combo antigen kits from field samples of domestic (broiler and layer chicken, native chicken) and semi-domestic (duck, goose, pigeon and quail) birds of Bangladesh. Samples were collected from naturally infected AI suspected domestic and semi-domestic birds of five different outbreak areas in Bangladesh. From each area two birds were selected for sampling, and from each bird three types of samples (tracheal, cloacal and oro-nasal swabs) were collected. A total of 210 field samples from a total of 70 birds were collected and tested using AI and NDV combo antigen rapid diagnostic kits in the study. All three different samples from a bird showed similar pattern of reaction. Out of 210 samples, 15 samples (5 birds), 63 samples (21 birds) and 27 samples (9 birds) were positive for AIV, NDV and both for AIV and NDV, respectively; whereas the remaining birds were negative for either AIV or NDV in this screening test. Among the five AIV positive, a layer chicken from wet market in Mymensingh, Netrokona, Gibandha and Kurigram and a native chicken from wet market in Kurigram area was positive to AIV. The semi-domestic birds are either positive to NDV or free from both AIV and NDV. This study revealed that the AIV and NDV rapid diagnostic kits could be effectively use to diagnose the respective virus in trachea, oro-nasal and cloacal samples simultaneously. AIV-NDV combo Ag test result clearly indicates that the test kit designed for AIV and NDV could diagnose the disease rapidly with less effort and higher scientific know how which could be used for the detection of AIV and NDV using field samples in large scale.

• Korean Journal of Veterinary Research
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• v.22 no.2
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• pp.175-185
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• 1982

Since bovine lymphosarcoma causes considerable economic loss to the dairy industry, seroepidemiological survey on bovine leucosis virus (BLV) was carried out for the dairy herds throughout the country to observe the epidemiological situation of the disease by using immunodiffusion test. Attempts were simultaneously made to detect bovine leucosis virus in the lymphocytes from BLV antibody-positive cattle by means of fluorescent antibody techniques, syncytium assay and electron microscopy. In immunodiffusion test for BLV antibody in 2003 heads of dairy cattle selected randomly from 164 herds, the prevalence of positive reactors by regions were 37.8% in Central, 27.2% in Honam (Southwest), 28.0% in Youngnam (Southeast) and 25.2% in Youngdong (East coast)and averaging 29.7%. By provinces, Chungcheong appeared the highest prevalence of BLV antibody carriers (41.8%), while Jeonbug revealed the lowest incidence rate (24.4%). When the results of serological studies were analyzed by age groups and the sizes of herds, the number of reactors increased gradually with the advance in the age of cattle and the herd size. The highest rate of BLV carriers was found in the ages between 6 and 8 years, and in the size of herds with 20 to 50 heads. One hundred and seventeen breeding bulls from the central regions were tested for BLV antibody. Four out of 70 bulls (5.7%) of Korean cattle and 14 out of 39 bulls (35.9%) of Holstein were reactive for BLV antigens. Of 164 dairy herds examined, 17 herds (10.4%) have no BLV antibody-positive cattle, while 42 herds (25.6%) were included in the range of 20 to 40% of the positive rate and 10 herds (6.1%) in the range of over 80% of the rate. When the lymphocytes from the BLV antibody carrying cattle were cultured in the presence of phytohemagglutinin and stained with FITC-conjugated sheep anti-BLV serum, 8 out of 11 cases (72.7%) of BLV positive cattle revealed specific fluorescence for BLV in the lymphocytes. In syncytium assay of the peripheral lymphocytes of the cattle, 5 out of 7 (71.4%) lymphocytes from BLV antibody carriers induced syncytia in the indicators of bovine embryonic splenic cells. The cultured lymphocytes were examined with an electron microscope to detect the BLV particles. Two out of 6 specimens (33.3%) from the reactors showed the typical type C virus with the size of 90 to 110 nm around microvilli and in intracytoplasmic vacuoles.

• Clinical and Experimental Pediatrics
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• v.48 no.12
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• pp.1348-1353
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• 2005

Purpose : Influenza is a respiratory disease which prevails widely every year and shows high morbidity and mortality among not only chronic invalids and the old, but also among infants and young children. To prevent community-acquired influenza infection, to facilitate prompt antiviral therapy and to avoid unnecessary use of antibiotics, an easy, rapid diagnostic method for the influenza virus is needed. We evaluated a lateral-flow immunoassay(QuickVue Influenza Test), compared to viral culture. Methods : During two consecutive years from Jan. 2004 to June 2004 and from Feb. 2005 to Jan. 2005, 408 patients who were suffering from fever, cough and/or sore throat and myalgia were enrolled in our study. A total of 408 patients were tested with $QuickVue^{(R)}$(Quidel Co., San Diego, USA) influenza rapid antigen test and virus cultures at the same time. Results : Of the 408 patients tested, children who showed positive results at the virus culture numbered 77; among them, 55(71.4 percent) were type A/H3N2 and 22(28.5 percent) were type B. QuickVue influenza test had a sensitivity of 71.4 percent and a specificity of 95.8 percent. The positive and negative predictive values were 79.7 percent and 93.5 percent, respectively. Conclusion : In our study, this test had comparable high sensitivity and high specificity and many advantages, such as being easy to perform and simple to interpret, and showing rapid results. If rapid influenza antigen tests are widely applied in the clinic, we can begin treatment more rapidly and reduce influenza complications and the abuse of antibiotics.