• Pediatric Infection and Vaccine
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• v.14 no.1
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• pp.69-74
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• 2007

Purpose : Human coronanviruses (hCovs) including hCoV-229E and hCoV-OC43 have been known as etiologic agents of the common colds and were regarded as clinically insignificant agents. However, recent identification of hCoV-NL63 and hCoV-HKU1 in children with lower respiratory tract infections has evoked the clinical concerns about their prevalence and the clinical significance of these hCoVs in children. This study was performed to investigate the prevalence of hCoVs in children with community-acquired pneumonia. Methods : From March 2006 to January 2007, nasopharyngeal specimens collected from children hospitalized with pneumonia, were tested for the presence of common respiratory viruses (respiratory syncytial virus, influenza A, influenza B, parainfluenza viruses, and adenovirus) using multiplex reverse transcriptase polymerase chain reaction (RT-PCR). Human metapneumovirus (hMPV) infection was excluded by nested RT-PCR using primers for the F-gene. To detect the different strains of hCoVs, nested RT-PCR assays specific for hCoVNL63, hCoV-OC43, hCoV-229E, and hCoV-HKU1 were performed. Results : Out of the 217 nasopharyngeal aspirate from children aged under 15 years, respiratory syncytial virus (RSV) was detected in 32 patients, hMPV in 18, human parainfluenza virus in 10, influenza virus A in 2, and adenovirus in 6. HCoVs were detected by RT-PCR in 8 (3.7%) of the 217 patients, hCoV-229E in 1, hCoV-NL63 in 3, and hCoVOC43 in 4 patients. HCoV-HKU1 was not detected in this study population. Conclusion : Recently identified hCoV-NL63 and hCoV-HKU1 seemed to have a little clinical significance in Korean children with severe or hospitalized community-acquired pneumonia.

• Proceedings of the Korean Vacuum Society Conference
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• 2013.08a
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• pp.88-89
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• 2013

A variety of influenza A viruses from animal hosts are continuously prevalent throughout the world which cause human epidemics resulting millions of human infections and enormous industrial and economic damages. Thus, early diagnosis of such pathogen is of paramount importance for biomedical examination and public healthcare screening. To approach this issue, here we propose a fully integrated Rotary genetic analysis system, called Rotary Genetic Analyzer, for on-site detection of influenza A viruses with high speed. The Rotary Genetic Analyzer is made up of four parts including a disposable microchip, a servo motor for precise and high rate spinning of the chip, thermal blocks for temperature control, and a miniaturized optical fluorescence detector as shown Fig. 1. A thermal block made from duralumin is integrated with a film heater at the bottom and a resistance temperature detector (RTD) in the middle. For the efficient performance of RT-PCR, three thermal blocks are placed on the Rotary stage and the temperature of each block is corresponded to the thermal cycling, namely $95^{\circ}C$ (denature), $58^{\circ}C$ (annealing), and $72^{\circ}C$ (extension). Rotary RT-PCR was performed to amplify the target gene which was monitored by an optical fluorescent detector above the extension block. A disposable microdevice (10 cm diameter) consists of a solid-phase extraction based sample pretreatment unit, bead chamber, and 4 ${\mu}L$ of the PCR chamber as shown Fig. 2. The microchip is fabricated using a patterned polycarbonate (PC) sheet with 1 mm thickness and a PC film with 130 ${\mu}m$ thickness, which layers are thermally bonded at $138^{\circ}C$ using acetone vapour. Silicatreated microglass beads with 150~212 ${\mu}L$ diameter are introduced into the sample pretreatment chambers and held in place by weir structure for construction of solid-phase extraction system. Fig. 3 shows strobed images of sequential loading of three samples. Three samples were loaded into the reservoir simultaneously (Fig. 3A), then the influenza A H3N2 viral RNA sample was loaded at 5000 RPM for 10 sec (Fig. 3B). Washing buffer was followed at 5000 RPM for 5 min (Fig. 3C), and angular frequency was decreased to 100 RPM for siphon priming of PCR cocktail to the channel as shown in Figure 3D. Finally the PCR cocktail was loaded to the bead chamber at 2000 RPM for 10 sec, and then RPM was increased up to 5000 RPM for 1 min to obtain the as much as PCR cocktail containing the RNA template (Fig. 3E). In this system, the wastes from RNA samples and washing buffer were transported to the waste chamber, which is fully filled to the chamber with precise optimization. Then, the PCR cocktail was able to transport to the PCR chamber. Fig. 3F shows the final image of the sample pretreatment. PCR cocktail containing RNA template is successfully isolated from waste. To detect the influenza A H3N2 virus, the purified RNA with PCR cocktail in the PCR chamber was amplified by using performed the RNA capture on the proposed microdevice. The fluorescence images were described in Figure 4A at the 0, 40 cycles. The fluorescence signal (40 cycle) was drastically increased confirming the influenza A H3N2 virus. The real-time profiles were successfully obtained using the optical fluorescence detector as shown in Figure 4B. The Rotary PCR and off-chip PCR were compared with same amount of influenza A H3N2 virus. The Ct value of Rotary PCR was smaller than the off-chip PCR without contamination. The whole process of the sample pretreatment and RT-PCR could be accomplished in 30 min on the fully integrated Rotary Genetic Analyzer system. We have demonstrated a fully integrated and portable Rotary Genetic Analyzer for detection of the gene expression of influenza A virus, which has 'Sample-in-answer-out' capability including sample pretreatment, rotary amplification, and optical detection. Target gene amplification was real-time monitored using the integrated Rotary Genetic Analyzer system.

• Korean Journal of Poultry Science
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• v.50 no.1
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• pp.41-50
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• 2023

Over the last decade, avian influenza (AI) has been considered an emerging disease that would become the next pandemic, particularly in countries like South Korea, with continuous animal outbreaks. In this situation, risk assessment is highly needed to prevent and prepare for human infection with AI. Thus, we developed the risk assessment matrix for a high-risk area of human infection with AI in South Korea based on the notion that risk is the multiplication of hazards with vulnerability. This matrix consisted of highly pathogenic avian influenza (HPAI) in poultry farms and the number of poultry-associated production facilities assumed as hazards of avian influenza and vulnerability, respectively. The average number of HPAI in poultry farms at the 229-municipal level as the hazard axis of the matrix was predicted using a negative binomial regression with nationwide outbreaks data from 2003 to 2018. The two components of the matrix were classified into five groups using the K-means clustering algorithm and multiplied, consequently producing the area-specific risk level of human infection. As a result, Naju-si, Jeongeup-si, and Namwon-si were categorized as high-risk areas for human infection with AI. These findings would contribute to designing the policies for human infection to minimize socio-economic damages.

• Pediatric Infection and Vaccine
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• v.24 no.1
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• pp.16-22
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• 2017

Purpose: We suspect there is a difference in the clinical manifestations and treatment response to antiviral drugs for influenza A and B. This study was conducted to investigate this difference. Methods: We collected information on pediatric patients, infected with the influenza virus, admitted to Dongguk University Ilsan Hospital from October 2013 to May 2015. We investigated the clinical manifestations of influenza and differences in treatment response to oseltamivir treatment for the two types of influenza. Results: A total of 138 patients were included. The mean age was $3.5{\pm}4.0$ years. When comparing the diseases associated with influenza A and B, croup (19.2% vs. 1.7%, P=0.001) was more common with influenza A infection. Myositis (0% vs. 6.7%, P=0.021) and gastroenteritis (29.5% vs. 46.7%, P=0.038) were more common with influenza B infection. When comparing the total fever duration from the start of oseltamivir administration, patients treated with oseltamivir within 2 days of fever had the shortest duration. Among the patients treated with oseltamivir, the duration of fever, after the start of oseltamivir treatment, for was shorter for influenza A infection than for influenza B infection ($16.0{\pm}19.1$ hours vs. $28.9{\pm}27.9$ hours, P=0.006). Conclusions: There appear to be differences in the accompanying diseases and antiviral medication responses between the two types of influenza. It is important to administer oseltamivir within 2 days of fever.

• Journal of Veterinary Science
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• v.21 no.2
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• pp.24.1-24.11
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• 2020

The pandemic of avian influenza viruses (AIVs) in Asia has caused enormous economic loss in poultry industry and human health threat, especially clade 2.3.4.4 H5 and H7 subtypes in recent years. The endemic chicken H6 virus in Taiwan has also brought about human and dog infections. Since wild waterfowls is the major AIV reservoir, it is important to monitor the diversified subtypes in wildfowl flocks in early stage to prevent viral reassortment and transmission. To develop a more efficient and sensitive approach is a key issue in epidemic control. In this study, we integrate multiplex reverse transcription recombinase polymerase amplification (RT-RPA) and capillary electrophoresis (CE) for high-throughput detection and differentiation of AIVs in wild waterfowls in Taiwan. Four viral genes were detected simultaneously, including nucleoprotein (NP) gene of all AIVs, hemagglutinin (HA) gene of clade 2.3.4.4 H5, H6 and H7 subtypes. The detection limit of the developed detection system could achieve as low as one copy number for each of the four viral gene targets. Sixty wild waterfowl field samples were tested and all of the four gene signals were unambiguously identified within 6 h, including the initial sample processing and the final CE data analysis. The results indicated that multiplex RT-RPA combined with CE was an excellent alternative for instant simultaneous AIV detection and subtype differentiation. The high efficiency and sensitivity of the proposed method could greatly assist in wild bird monitoring and epidemic control of poultry.

• Pediatric Infection and Vaccine
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• v.21 no.3
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• pp.207-213
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• 2014

Purpose: This study aimed to investigate the association between respiratory virus infection and pneumococcal colonization in children. Methods: From May 2009 to June 2010, nasopharyngeal (NP) aspirates were obtained from patients under 18 years old who visited Seoul National University Children's Hospital for respiratory symptoms. NP samples were used to detect respiratory viruses (influenza virus A and B, parainfluenza virus 1, 2 and 3, respiratory syncytial virus A and B, adenovirus, rhinovirus A/B, human metapneumovirus, human coronavirus 229E/NL63 and OC43/HKU1) by RT-PCR and pneumococcus by culture. Results: Median age of the patients was 27 months old. A total of 1,367 NP aspirates were tested for respiratory viruses and pneumococcus. Pneumococcus was isolated from 228 (16.7%) of samples and respiratory viruses were detected from 731 (53.5%). Common viruses were rhinovirus (18.4%), respiratory syncytial virus (RSV) A (10.6%), adenovirus (6.9%), influenza virus A (6.8%). Pneumococcal isolation rate was significantly higher in the cases of positive virus detection than negative detection [21.3% (156/731) vs. 11.3% (72/636), P <0.001]. For individual viruses, pneumococcal isolation rate was positively associated with detection of influenza virus A [24.7% (23/93) vs 16.1% (205/1274), P=0.001], RSV A [28.3% (41/145) vs 15.3% (187/1222), P=0.001], RSV B [31.3% (10/32) vs 16.3% (218/1335), P=0.042], rhinovirus A/B [22.6% (57/252) vs 15.3% (171/1115), P=0.010]. Conclusion: The study revealed that pneumococcal isolation from NP aspirates is related with respiratory virus detection. The result of this study could be used to investigate how respiratory viruses and pneumococcus cause clinical diseases.

• Journal of Microbiology and Biotechnology
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• v.25 no.2
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• pp.280-287
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• 2015

Current influenza vaccines are produced in embryonated chicken eggs. However, egg-based vaccines have various problems. To address these problems, recombinant protein vaccines have been developed as new vaccine candidates. Unfortunately, recombinant proteins frequently encounter aggregation and low stability during their biogenesis. It has been previously demonstrated that recombinantly expressed proteins can be greatly stabilized with high solubility by fusing stabilizing peptide (SP) derived from the C-terminal acidic tail of human synuclein (ATS). To investigate whether SP fusion proteins can induce protective immunity in mice, we produced influenza HA and SP fusion protein using a baculovirus expression system. In in vitro tests, SP-fused recombinant HA1 (SP-rHA1) was shown to be more stable than recombinant HA1 (rHA1). Mice were immunized intramuscularly with baculovirus-expressed rHA1 protein or SP-rHA1 protein ($2{\mu}g/mouse$) formulated with aluminum hydroxide. Antibody responses were determined by ELISA and hemagglutination inhibition assay. We observed that SP-rHA1 immunization elicited HA-specific antibody responses that were comparable to rHA1 immunization. These results indicate that fusion of SP to rHA1 does not negatively affect the immunogenicity of the vaccine candidate. Therefore, it is possible to apply SP fusion technology to develop stable recombinant protein vaccines with high solubility.

• Journal of Multimedia Information System
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• v.6 no.4
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• pp.293-302
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• 2019

In the past, there was a theory that influenza wasn't transmitted directly from birds but was infected to humans via swains. Recently, molecular level research has progressed, and it was confirmed that the avian influenza virus can directly infected to human lung and intestinal epithelial cells. Three pandemicsin the past 100 years were also infected to humans directly from birds. In view of such scientific background, we are developing a method for screening sick birds by monitoring the physiological characteristics of birds in a contactless manner with sensors. Here, the movement of respiratory muscles and abdominal muscles under autonomic innervation was monitored using a magnet and Hall sensor sewn on the thoracic wall, and other multimedia devices. This paper presents and discusses the results of experiments involving continuous periodic noise discovered during flight experiments with a data logger mounted on a Japanese pheasant from 2012 to 2015. A brief summary is given as the below: 1. Magnet and Hall sensor sewn to the left and right chest walls, bipolar electrocardiograms between the thoracic walls, posterior thoracic air sac pressure, angular velocity sensors sewn on the back and hips, and optical reflection of LEDs (blue and green) from the skin of the hips allow observation of periodic vibrations(fasciculations) in the waves. No such analysis has been reported before. 2. These fasciculations are presumed to be derived from muscle to maintain and control air sac pressure. 3. Since each muscle fiber is spatially Gaussian distributed from the sympathetic nerve, the envelope is assumed to plot a Gaussian curve. 4. Since avian trunk muscles contract periodically at all time, we assume that the sympathetic nerve dominates in their control. 5. The technique of sewing a magnet to the thoracic wall and measuring the strength of the magnetic field with a Hall sensor can be applied to screen for early stage of avian influenza, with a sensor attached to the chicken enclosure.

• Genomics & Informatics
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• v.14 no.3
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• pp.96-103
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• 2016

The influenza A (H1N1) virus, also known as swine flu is a leading cause of morbidity and mortality since 2009. There is a need to explore novel anti-viral drugs for overcoming the epidemics. Traditionally, different plant extracts of garlic, ginger, kalmegh, ajwain, green tea, turmeric, menthe, tulsi, etc. have been used as hopeful source of prevention and treatment of human influenza. The H1N1 virus contains an important glycoprotein, known as neuraminidase (NA) that is mainly responsible for initiation of viral infection and is essential for the life cycle of H1N1. It is responsible for sialic acid cleavage from glycans of the infected cell. We employed amino acid sequence of H1N1 NA to predict the tertiary structure using Phyre2 server and validated using ProCheck, ProSA, ProQ, and ERRAT server. Further, the modelled structure was docked with thirteen natural compounds of plant origin using AutoDock4.2. Most of the natural compounds showed effective inhibitory activity against H1N1 NA in binding condition. This study also highlights interaction of these natural inhibitors with amino residues of NA protein. Furthermore, among 13 natural compounds, theaflavin, found in green tea, was observed to inhibit H1N1 NA proteins strongly supported by lowest docking energy. Hence, it may be of interest to consider theaflavin for further in vitro and in vivo evaluation.

• Journal of the Korea Convergence Society
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• v.12 no.4
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• pp.201-206
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• 2021

Damage to the highly pathogenic avian influenza virus(H5N1) continues to increase, but there is a lack of antiviral research. In this study, we analyze antiviral properties on H5N1 by coating Cu/TiO2 photocatalyst on polyethylene films. The specimen was manufactured a photocatalyst master batch and coated both sides of the 3-layer polyethylene fabric at 280℃ from the extrusion coating machine. The results showed a 99.9% decrease in the Staphylococcus aureus and Escherichia coli. In particular, H5N1 type highly pathogenic avian influenza viruses, which is capable of human infection, has been found to decrease 99.9% within five minutes of contact with Cu/TiO2 films. Antibacterial effects of films coated with photocatalyst are known, but this study also confirmed the antiviral effects.