Shared Attention Network와 Multi-Query Attention을 이용한 Transformer 모델의 디코더 속도 개선

최민주(Minjoo Choi),김도경(Dokyoung Kim),황현선(Hyunsun Hwang),이창기(Changki Lee),이성민(Seongmin Lee),류우종(Woojong Ryu),염홍선(Hongseon Yeom),김준석(Junseok Kim) 한국정보과학회 2021 한국정보과학회 학술발표논문집 Vol.2021 No.6

Efficient antiviral co-delivery using polymersomes by controlling the surface density of cell-targeting groups for influenza A virus treatment

Chun, Haejin,Yeom, Minjoo,Kim, Hyun-Ouk,Lim, Jong-Woo,Na, Woonsung,Park, Geunseon,Park, Chaewon,Kang, Aram,Yun, Dayeon,Kim, Jihye,Song, Daesub,Haam, Seungjoo The Royal Society of Chemistry 2018 Polymer chemistry Vol.9 No.16

<P>Influenza A virus (IAV), which causes one of the most contagious diseases, is a global health concern and is responsible for seasonal epidemics and pandemics. Despite notable efforts towards developing antiviral agents and drugs, a vast majority of these, especially intracellular drugs, have shown limited efficacy due to non-specificity and low viability under physiological or endosomal conditions. Polymersomes consist of phenylboronic acid (PBA) pendant group polymers (PBASomes) and can act as drug carriers; they have sialic acid-targeting properties and can gain greater access to the intracellular space for the transport of antivirals within the host cell. Amphiphilic copolymers comprising methoxy-poly(ethylene glycol)-<I>block</I>-poly(phenylalanine) (mPEG-<I>b</I>-pPhe) formed polymersomes, which encapsulated mir-323a in the core and favipiravir in the exterior layer as hydrophilic and hydrophobic antivirals, respectively. For maximizing the cellular uptake of PBASomes <I>via</I> receptor-mediated endocytosis, the surface density of PBA was controlled with PBA-functionalized copolymers (PBA-PEG-pPhe). Combination therapy by employing polymersomes with PBA functional groups induced a synergistic effect against H1N1 virus infection <I>in vitro</I>. We believe that antiviral co-delivery using these polymersomes would provide better opportunities to improve transfection of therapeutic substances for IAV treatment.</P>

Recombinant influenza virus expressing naturally truncated NS gene from H3N8 equine influenza virus confer self-attenuation for live viral vaccine application

( Woonsung Na ),( Minjoo Yeom ),( Sun-woo Yoon ),( Aram Kang ),( Huijun Yuk ),( Daesub Song ) 대한인수공통전염병학회 2016 창립총회 및 학술대회 초록집 Vol.2016 No.1

Seasonal and pandemic influenza viruses have threatened global health and caused tremendous economic damage. Inactivated influenza virus vaccines (IIV) only elicit a homologous hemagglutinin, which protect narrow spectrum of influenza virus strains. To overcome the limitation, broadly protecting influenza vaccines are needed utilizing a conserved region of antigen, and epitopes that can evoke immune responses with CD8 T lymphocytes(CTL).The live attenuated influenza virus vaccine (LAIV) with intranasal administration can elicit specific CTL and mucosal serum antibody which cross-protects a wide range of influenza viruses. Here we utilized the H3N8 equine influenza virus that has partially deleted NS gene, which infect into host only one round without pathogenicity. This live EIV vaccine inoculated nasally elicits more effective protection than inactivated EIV vaccine with intramuscular injection, making promising candidate as EIV live attenuated vaccine.

Genetic variations of PEDV of field isolates from Vietnam and plaque selected viruses after 20 serial passages in Vero cells

( Thi Thu Hang Vu ),( Minjoo Yeom ),( Woonsung Na ),( Van Phan Le ),( Daesub Song ) 대한인수공통전염병학회 2019 창립총회 및 학술대회 초록집 Vol.2019 No.1

Introduction: The antigenic diversity makes porcine epidemic diarrhea (PED) as a challenge for the pig industry in all over the world. The emergence of PED virus (PEDV) causes significantly economic loss in pig farms annually. In this research, as an initial step, full-length of spike (S) gene of 8 PEDV-positive samples collected from the farms and 16 clones of 20th passage- PEDV2 strain were sequenced and analyzed to identify the genetic variation of PEDV in the field as well as in the cell adapted condition. Methods: 8 positive PEDV samples were collected from outbreaks in Vietnam during 2018. While PEDV2 strain after 20 serial passages in Vero cell was plaque purified, 16 clones were selected and propagated by transferring from the plaque assay plate into the new plates and then to the T25 tissue culture flasks. RNA of all viral samples were purified and used as the template of RT-PCR with S gene specific primers for sequencing. BioEdit and MEGA 6.0 program were used for sequence analysis. Results: 8 PEDV strains circulating in Vietnam during 2018 belong to three different clades as Asian non-S INDEL, new S INDEL and classical S INDEL. Otherwise 16 PEDV clones from 20^th passage- PEDV2 strain were also divided into three groups with different deletion inside S gene sequence compared with original strain. Conclusion: PED virus strains have variable ability in various environmental conditions, in the field as well as in cell culture. From this result, in each outbreak, samples should be classified independently so that a proper vaccine solution can be developed. Further studies need to be done to clarify the relationship between genetic variants and pathogenicity of PED virus strains.

Mouse adaptation of recombinant canine influenza increase pathogenicity H3N2 virus in mice

( Aram Kang ),( Woonsung Na ),( Minjoo Yeom ),( Daesub Song ) 대한인수공통전염병학회 2019 창립총회 및 학술대회 초록집 Vol.2019 No.1

Introduction: Canine H3N2 influenza virus (CIV H3N2) of avian-origin was first isolated and found to induce acute respiratory disease in South Korea dogs in 2007. The virulence of a novel CIV H3N2 recombinant, VC378, which was CIV H3N2 recombinant derived from pandemic (pdm) H1N1 and CIV H3N2 in companion animals was demonstrated in mice. That was notable because mice inoculated with an equivalent dose of classical CIV H3N2 were shown low or no-pathogenic. However, our experiment showed that the inoculation of VC378 CIV in mice still showed a low pathogenic. We evaluated the pathogenic potential that are possibly associated with the adaptation of CIV H3N2 and VC378 virus to mice. Materials and methods: The recombinant CIV strain A/canine/Korea/01-VC378/2012 (H3N2), and the CIV H3N2 A/Canine/Korea/01/2007 (H3N2) were used. C57/BL6 mice (10 weeks old) were inoculated intranasally with virus and mouse-adaptation was performed through 10 lung-to-lung passages. The 10th passaged viruses were grown in the allantoic cavity of 11-day-old embryonated chicken eggs. The 50% mouse lethal dose (MLD50) was measured using groups of C57/BL6 mice (10 weeks old). The MLD50 values were calculated by the Reed and Muench (1938) method after a 14-day observation period and are expressed as EID50. Results: The parental CIV H3N2 virus MLD50 was >7.5 log10 EID50, whereas the MLD50 of the CIV H3N2 mouse-adapted virus was 5.5 log10 EID50. The VC378 virus MLD50 (8.26 log10 EID50) also reduced compared to the parental VC378 virus MLD50 (5 log10 EID50). The MLD50 of the mouse-adapted viruses were reduced 100-2000fold when compared to the parental virus. Conclusion: We confirmed that no or low pathogenicity of canine influenza virus H3N2 become highly pathogenicity after serial passage infection in the mice model.

Sputum processing method for rapid diagnosis of Middle East respiratory syndrome coronavirus (MERS-CoV)

( Aram Kang ),( Woonsung Na ),( Minjoo Yeom ),( Hyekwon Kim ),( Sun-woo Yoon ),( Heejun Yook ),( Dae-gwin Jeong ),( Daesub Song ) 대한인수공통전염병학회 2017 창립총회 및 학술대회 초록집 Vol.2017 No.1

Introduction: Middle East respiratory syndrome coronavirus (MERS-CoV), a member of the family of betacoronavirus, was first identified in Saudi Arabia in 2012. MERS-CoV has ability to cross the host species from camel to human causing severe acute respiratory illnesses, and spread by contact in human population. For diagnosis of MERS-CoV in camels, the immunochromatographic assay (ICA) has been used due to its rapid decision and prompt triage of infected animal for the early quarantine. However, when the ICA is applied to an expectorated sputum in which antigens are present, the viscosity of sputum interferes with the migration of the antigens on the test strip. To overcome this limitation, it is necessary to use a mucolytic agent without affecting the antigens. In this study, we have developed a sputum pre-treatment method by testing specimens of the sputa spiked with alphacorona virus and MERS-CoV. Methods: Two mucolytic agents were used: Tris(2-carboxyethyl) Phosphine (TCEP) and N-acetyl-L-cysteine (NALC) and prepared at various concentration to treat with sputum. Bovine serum albumin (BSA) was used as a blocking agent and protease inhibitor cocktail (PI) was used to inhibit the cleavage of the antigens. After treating the compound to sputum, the mixture was applied to colloidal gold-based immunochromatographic test strip for rapid detection of MERS-CoV or alpha coronavirus (BIONOTE Inc., South Korea). The intensities of colloidal gold were measured by MEDISENSOR Gold reader (SD BIOSENSOR Ltd., South Korea). Results: Intensity of test line was higher when the sputa spiked with alpha coronavirus was processed with TCEP, BSA and PI than with TCEP alone. In the case of the sputa spiked with inactivated MERS-CoV, mixture of TCEP and BSA presented higher intensities, while it decreased in the addition of PI. This was reproduced when compound of NALC and BSA was treated to the specimens. Conclusion: In this study, we demonstrated that mixture of the mucolytics, blocking agent and protease inhibitor together effectively dissolve the viscosity of sputum minimizing the effect on antigens, which is more suitable for use in flow immunochromatographic test kit than when sputum or mucolytics alone were used.

DPO based Multiplex PCR for detecting pathogens causing canine infectious respiratory disease and its usefulness for analyzing epidemiology of canine influenza H₃N₈, canine H₃N₂, pandemic H1N1

( Heejun Yook ),( Woonsung Na ),( Minjoo Yeom ),( Aram Kang ),( Daesub Song ) 대한인수공통전염병학회 2017 창립총회 및 학술대회 초록집 Vol.2017 No.1

Introduction: Canine infectious respiratory disease (CIRD) is one of the most common contagious disease and spread worldwide in dog population. Numerous bacteria species and viruses contribute CIRD with synergistic co-infection in host leading clinical symptoms of flu-like illnesses such as dry coughing and sneezing, which alone limits the differential diagnosis. To provide an appropriate treatment by differential diagnosis, it is required that high sensitive and specific diagnostic system to detect various pathogens simultaneously. In this study, we have designed novel multiplex PCR(mPCR) for differential diagnosis of threatening pathogens that cause CIRD: Bordetella bronchiseptica, canine distemper virus(CDV), canine influenza virus H3N8 (cH3N8), canine influenza H3N2 (cH3N2), and pandemic influenza virus H1N1 (pH1N1). Methods: Dual priming oligonucleodtide(DPO) system was used, in which contains stabilizing priming region, determining region and polydeoxyinosine linker, to enhance sensitivity and specificity by reducing non-specific amplicon. Initial sequence analysis and securing the conservancy for the sequences were conducted using Bio Edit ver. 7.0, and primers for mPCR were designed using Primer-BLAST (NCBI) Results: The five different pathogens of CIRD were discriminated by 100bp size difference of each PCR product according to their target sequence size in gel electrophoresis, which indicates specific primer pairs amplifies target template, respectively. Various annealing temperature were applied and the optimal reaction was determined to 57℃. Conclusion: In this study, we have developed a DPO based novel multiplex PCR that discriminate five CIRD pathogens concurrently. This provides exact diagnosis of CIRD and supports an appropriate treatment to infected dogs. Especially, the mPCR can differentiate three kinds of influenza viruses to which dogs are susceptible, and this could be useful tool for surveillance study of influenza viruses in dog population. Therefore, this multiplex PCR can offer powerful diagnosis tool and effectively help both treatment and surveillance of CIRD.

Dengue virus-Polymersome Hybrid nanovesicles for Real-Time Single Virus Tracking

( Hyun-ouk Kim ),( Woonsung Na ),( Minjoo Yeom ),( Seungjoo Haam ),( Daesub Song ) 대한인수공통전염병학회 2019 창립총회 및 학술대회 초록집 Vol.2019 No.1

Introduction: Dengue virus is one of the major infectious human pathogens worldwide. Currently, no antiviral drug has become available against the dengue virus induced diseases since little is known regarding how dengue virus interacts with host cells. Dengue virus-polymersome hybrid nanovesicles are powerful tools to explain the dynamics of host cell-virus interaction and tracking. Methods: Dengue virus-polymersome hybrid nanovesicles were prepared by dialysis method. We showed that DiD and BODIPY FL C5-Ceramide encapsulated dengue virus-polymersome hybrid nanovesicles triggered the formation of red and green fluorescence by using real-time fluorescence microscopy. The internalization to escape endosomal entrapment was determined by confocal laser scanning microscopy. Results: Fluorescence images of DiD and BODIPY FL C5-Ceramide encapsulated dengue virus-polymersome hybrid nanovesicles illustrated at different time points. For real-time virus tracking study, it is desirable to acquire images to obtain adequately high resolution to monitor the dynamics of host cell-virus interaction and tracking in living cells. Conclusion: This study exploited a virus-nanovesicles tracking technology to approach whether dengue virus interacts with autophagy. Therefore, we demonstrated that dengue virus-polymersome hybrid nanovesicles will be utilized for virus tracking studies to examine the mechanisms of viral infections.

Virulence of a novel reassortant canine H3N2 influenza virus in ferret, dog and mice models

( Woonsung Na ),( Kwang-soo Lyoo ),( Minjoo Yeom ),( Dae-gwin Jeong ),( Daesub Song ),( Chang-ung Kim ),( Jeong-ki Kim ),( Daesub Song ) 대한인수공통전염병학회 2016 창립총회 및 학술대회 초록집 Vol.2016 No.1

The outbreak of a canine influenza virus (CIV) H3N2 reassortanted from pandemic (pdm) H1N1 and CIV H3N2 in companion animals has underscored the urgent need to monitor CIV infection for potential zoonotic transmission of influenza viruses to humans. In this study, we assessed the virulence of a novel CIV H3N2 (VC378) from a pdm H1N1 and CIV H3N2 coinfected dog in ferrets, dogs, and mice. Significantly enhanced virulence of VC378 was demonstrated in mice, although the transmissibility and pathogenicity of VC378 were similar to those of classic H3N2 in ferrets and dogs. This is notable because mice inoculated with an equivalent dose of classic CIV H3N2 showed no clinical signs and no lethality. We found that the PA and NS gene segments of VC378 were introduced from pdmH1N1, and these genes included amino acid substitutions; PAP224S and NS-I123V, that were previously found to be associated with virulence enhancement in mice. Thus, we speculate that the natural reassortment of CIV between pdm H1N1 and CIV H3N2 can confer virulence and that continuous surveillance is needed to monitor the evolution of CIV in companion animals.

Genetic Characteristics and Immunogenicity of Pandemic H1N1 Influenza Virus Isolate from Pig in Korea

Moon, Hyoung Joon,Oh, Jin Sik,Na, Woonsung,Yeom, Minjoo,Han, Sang Yoon,Kim, Sung Jae,Park, Bong Kyun,Song, Dae Sub,Kang, Bo Kyu 한국조명·전기설비학회 2016 한국조명·전기설비학회 학술대회논문집 Vol. No.

<P>A pandemic influenza A (H1N1) virus strain was isolated from a pig farm in Korea in December 2009. The strain was propagated in and isolated from both the Madin-Darby canine kidney cell line and embryonated eggs. The partial and complete sequences of the strain were identical to those of A/California/04/2009, with >99% sequence similarity in the HA, NA, M, NS, NP, PA, PB1, and PB2 genes. The isolated strain was inactivated and used to prepare a swine influenza vaccine. This trial vaccine, containing the new isolate that has high sequence similarity with the pandemic influenza A (H1N1) virus, resulted in seroconversion in Guinea pigs and piglets. This strain could therefore be a potential vaccine candidate for swine influenza control in commercial farms.</P>