• 제목/요약/키워드: virus particle

검색결과 108건 처리시간 0.027초

Preparation of Microspheres Encapsulating a Recombinant TIMP-1 Adenovirus and their Inhibition of Proliferation of Hepatocellular Carcinoma Cells

  • Xia, Dong;Yao, Hui;Liu, Qing;Xu, Liang
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권12호
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    • pp.6363-6368
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    • 2012
  • Objective: The study aim was to prepare poly-DL-lactide-poly (PELA) microspheres encapsulating recombinant tissue inhibitors of metalloproteinase-1 (TIMP-1) in an adenovirus to investigate its inhibition on the proliferation of hepatocellular carcinoma cells HepG2. Methods: Microspheres were prepared by encapsulating the recombinant TIMP-1 adenovirus into biodegradable PELA. The particle size, viral load, encapsulation efficiency and in-vitro release were measured. Microspheres were used to infect HepG2 cells, then infection efficiency was examined under a fluorescent microscope and ultrastructural changes assessed by TEM. Expression of TIMP-1 mRNA in HepG2 cells was examined by semi-quantitative RT-PCR and proliferation by MTT and cell growth curve assays. Results: We successfully prepared microspheres encapsulating recombinant TIMP-1 adenovirus with a diameter of $1.965{\mu}m$, an encapsulation efficiency of 60.0%, a viral load of $10.5{\times}10^8/mg$ and approximate 60% of virus release within 120 h, the total releasing time of which was longer than 240 h. The microspheres were confirmed to be non-toxic with blank microspheres. Infected HepG2 cells could stably maintain in-vitro expression of TIMP-1, with significantly effects on biological behaviour Conclusion: PELA microspheres encapsulating a recombinant TIMP-1 adenovirus can markedly inhibit the proliferation of HepG2 cells, which provides an experimental basis for polymer/chemistry-based gene therapy of hepatocellular carcinomas.

음이온성 유화제로 수식된 폴리락티드/글리코리드 공중합체 나노 입자와 백신의 결합성 (Binding of Vaccine and Poly(DL-lactide-co-glycolide) Nanoparticle Modified with Anionic Surfactant)

  • 최민수;박은석;지상철;신병철
    • Journal of Pharmaceutical Investigation
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    • 제34권3호
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    • pp.177-183
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    • 2004
  • Recently, studies on intranasal mucosa delivery of influenza vaccine have been actively developed because of lack of pain and ease of administration. We studied on preparation of nanoparticle delivery system using biodegradable polymer as a poly(DL-lactide-co-glycolide) (PLGA) and their binding characteristics with vaccine. Three kinds of PLGA nanoparticles were prepared by spontaneous emulsification solvent diffusion (SESD) method using sodium dodecyl sulfate and sodium laurate as an anionic surfactant and Lutrol F68 (polyethylene glycol-block-polypropylene glycol copolymer) as a nonionic surfactant. The 5-aminofluorescein labeled vaccine was coated on the surface of nanoparticles by ionic complex. The complexes between vaccine and nanoparticles were confirmed by change of the size. After vaccine coating on the surface of anionic nanoparticles, particle size was increased from 174 to 1,040 nm. However the size of nonionic nanoparticles was not more increased than size of anionic nanoparticles. The amount of coated vaccine on the surface of PLGA nanoparticles was $14.32\;{\mu}g/mg$ with sodium dodecyl sulfate, $12.41\;{\mu}g/mg$ with sodium laurate, and $9.47{\mu}g/mg$ with Lutrol F68, respectively. In conclusion, prepared nanoparticles in this study is possible to use as a virus-like nanoparticles and it could be accept in the field of influenza vaccine delivery system.

Comparison of Immune Responses to the PCV2 Replicase-Capsid and Capsid Virus-Like Particle Vaccines in Mice

  • Jung, Bo-Kyoung;Kim, Hye-Ran;Lee, Young-Hyeon;Jang, Hyun;Chang, Kyung-Soo
    • Journal of Microbiology and Biotechnology
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    • 제29권3호
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    • pp.482-488
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    • 2019
  • Porcine circovirus type 2 (PCV2) is the causative agent of postweaning multisystemic wasting syndrome (PMWS) in pigs. Replicase (Rep) proteins are considered essential for viral replication. Capsid (Cap) protein is the primary immunogenic protein that induces protective immunity. Little is known about comparison on the immunogenicity of PCV2 Rep and Cap fusion protein and Cap protein. In the present study, recombinant baculoviruses expressing the Rep-Cap fusion protein (Bac-Rep-Cap) and the Cap protein (Bac-Cap) of PCV2 were constructed and confirmed with western blot and indirect fluorescence assay. Immunogenicities of the two recombinant proteins were tested in mice. The titers of antibodies were determined with a PCV2-specific enzyme-linked immunosorbent assay (ELISA) and a serum neutralization assay. The $IFN-{\gamma}$ response of immunized mice was measured by ELISA. The mice immunized with the Bac-Rep-Cap and Bac-Cap successfully produced Cap-specific immunoreaction. The mice immunized with the Bac-Cap developed higher PCV2-specific neutralizing antibody titers than mice injected with the Bac-Rep-Cap. $IFN-{\gamma}$ in the Bac-Rep-Cap group was increased compared to those in the Bac-Cap group. Vaccination of mice with the Bac-Rep-Cap showed significantly decreased protective efficacy compared to the Bac-Cap. Our findings will indubitably not only lead to a better understanding of the immunogenicity of PCV2, but also improved vaccines.

Glycogen Synthase Kinase-3 Interaction Domain Enhances Phosphorylation of SARS-CoV-2 Nucleocapsid Protein

  • Jun Seop, Yun;Hyeeun, Song;Nam Hee, Kim;So Young, Cha;Kyu Ho, Hwang;Jae Eun, Lee;Cheol-Hee, Jeong;Sang Hyun, Song;Seonghun, Kim;Eunae Sandra, Cho;Hyun Sil, Kim;Jong In, Yook
    • Molecules and Cells
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    • 제45권12호
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    • pp.911-922
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    • 2022
  • A structural protein of SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2), nucleocapsid (N) protein is phosphorylated by glycogen synthase kinase (GSK)-3 on the serine/arginine (SR) rich motif located in disordered regions. Although phosphorylation by GSK-3β constitutes a critical event for viral replication, the molecular mechanism underlying N phosphorylation is not well understood. In this study, we found the putative alpha-helix L/FxxxL/AxxRL motif known as the GSK-3 interacting domain (GID), found in many endogenous GSK-3β binding proteins, such as Axins, FRATs, WWOX, and GSKIP. Indeed, N interacts with GSK-3β similarly to Axin, and Leu to Glu substitution of the GID abolished the interaction, with loss of N phosphorylation. The N phosphorylation is also required for its structural loading in a virus-like particle (VLP). Compared to other coronaviruses, N of Sarbecovirus lineage including bat RaTG13 harbors a CDK1-primed phosphorylation site and Gly-rich linker for enhanced phosphorylation by GSK-3β. Furthermore, we found that the S202R mutant found in Delta and R203K/G204R mutant found in the Omicron variant allow increased abundance and hyper-phosphorylation of N. Our observations suggest that GID and mutations for increased phosphorylation in N may have contributed to the evolution of variants.

참굴 채묘 부진 원인 구명에 관한 연구 -병리 발생학적 조사를 통한 참굴 모패의 건강도와 난 및 유생의 생존율과의 상관관계 구명 - (Investigation on the Cause of Bad Natural Seed Collection of the Pacific Oyster, Crassostrea gigas: Relationships between the Conditions of Mother Shell and the Viability of the Released Eggs and Larvae Based on the Pathological and Embryological Survey)

  • 박미선;류호영;이태식
    • 한국수산과학회지
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    • 제32권1호
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    • pp.62-67
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    • 1999
  • 1992, 1993년에 경남 통영, 거제 및 전남 여수 등 남해안 주요 굴 채묘 어장에서 참굴, Crassostrea gigas의 극심한 채묘 부진 현상이 발생하여 이에 대한 원인 구명의 일환으로 각 시험 어장 서식 모패 중의 난소기생충 감염율을 조사하고 감염난 및 비감염난에 대한 발생 비교 시험을 실시하였다. 그리고, 모패 중 난의 건강도 판정 및 발생 가능성 추정을 위하여 난 지질 함량을 측정하였다. 또한, 각 채묘 부진 해역에서 채집한 굴 유생 중의 세균의 오염정도와 오염균의 세균상 등을 조사하였다. 1992년 8월에서 1993년 9월 사이 경남 통영 저산, 거제 오수 및 전남 여수 경도, 굴전 지역 굴 암컷 모패의 난소기생충, Marteiliodes chungmuensis 감염 율은 각각 $11.8\~100\%,\;14.3\~100\%,\;$15.4\~93.3\%,\;12.5\~91.7\%$였으나, 충남 대천의 자연산 굴 암컷 모패에서는 난소기생충이 검출되지 않았다. 한편, 난소기생충에 감염된 난의 세포질 내에서는 직경 78$\~$80nm의 바이러스성 입자 (virus-like particle)가 관찰되었다. 난소기생충에 감염된 난을 사용한 인공 발생 실험에서 기생충에 감염된 난은 수정이 불가능하였으며, 동일 생식소에 감염난과 함께 있었던 정상난도 수정은 가능하였으나, $80\%$ 이상이 형태적으로 비정상적인 발생을 하였고, 정상적으로 발생하여 D형 및 초기각정기까지 도달한 유생도 외부 각부터 녹기 시작하여 연체부까지 파괴되는 세포 괴사 현상을 나타내며 전량 폐사하였다. 채묘 부진 현상이 발생한 통영 저산 양식산 참굴의 난지질 함량은 3.8ng/egg이었으나, 충남 대천 자연산의 난지질은 6.2ng/egg였다. 한편, 발생 전 낮은 난 지질 함량을 나타내었던 통영 저산 산의 난은 정상적인 수정, 발생 단계를 거쳐 수정 7일째 초기각정기까지는 발달하였으나, 이후 발생이 진전되지 못한 채 수정 10일째 전량 폐사하였다. 각 시험어장에서 채취한 참굴 유생의 단위개체당 생균수는 거제 오수가 8,100으로 가장 높았고, 통영 저산이 98, 여수 굴전이 5.5로 나타났다. 각 시험어장에서 채수한 해수 중의 생균수는 여수 굴전이 14,000/ml으로 가장 높았으며, 거제 오수와 통영 저산이 각각 370, 260/ml으로 비슷하였다. 채묘 부진 해역에서 채집된 패류 유생 중의 세균상은 Pseudomonas속 및 그 유사세균이 $53.3\~87.1\%$로 절대 우점종을 차지하였다.

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Development of dry milling suitable rice cultivar to invigorate rice processing products

  • Jeung, Ji-Ung
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2017년도 9th Asian Crop Science Association conference
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    • pp.10-10
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    • 2017
  • Rice consumption has been continuously decreasing as the eating habits of Koreans have become westernized and diversified. The per capita annual rice consumption in Korea has dropped sharply from 136.4 kg in 1970 to 61.9 kg in 2016. The Korean government, therefore, has been trying to promote rice consumption by invigorating the processed food industry using rice flour. To facilitate the market for processed rice foods, it is essential to develop proper milling technology in terms of flour particle size and damaged starch content to produce high quality rice flour at competitive cost. Dry milling and wet milling are the two major processes used to produce rice flour. Although the dry milling process is relatively simple with a lower production cost, damaged starch content increases because of the high grain hardness of rice. In wet milling, the quality of rice flour is improved by reducing flour particle size as well as damaged starch content through soaking procedures. However, the production costs are high because of the additional expenses associated with the disposal of waste water, sterilization and drying of the wet flour. Recently developed technologies such as jet milling and cryogenic milling also require expensive investment and production. Therefore, developing new rice cultivars with dry milling adaptability as well as good processing properties is an important goal of rice breeding in Korea. 'Suweon 542' is a floury endosperm mutant line derived from sodium azide treatment on a high-yield, early maturing, and non-glutinous japonica rice cultivar, 'Namil'. Compared with the wild type, after dry milling process, the grain hardness of 'Suweon 542' was significantly lower because of its round and loosely packed starch granules. Also, the flour of 'Suweon 542' had significantly smaller particles and less damaged starch than 'Namil' and other rice cultivars and its particle size distribution was similar to a commercial wheat cultivar. Recently, through collaborations with nine universities and food companies, a total of 21 kinds of processed prototypes, using the dry milling flour of 'Suweon 542', were evaluated. In the production of major rice processing products, there was no significant quality difference between the flours prepared by wet milling and dry milling. Although the amount of water added to the dough was slightly increased, it was confirmed that the recipe applying the wet flour could be used without significant change. To efficiently transfer the floury endosperm characteristics of 'Suweon 542' to other commercial rice cultivars, it is essential to develop DNA marker tightly linked to the target gene. Association analysis using 70 genome-wide SSR markers and 94 F2 plants derived from 'Suweon 542'/'Milyang 23' showed that markers on chromosome 5 explained a large portion of the variation in floury grains percentage (FGP). Further analysis with an increased number of SSR markers revealed that the floury endosperm of 'Suweon 542' was directed by a major recessive locus, flo7(t), located in the 19.33-19.86 Mbp region of chromosome 5, with RM18639 explaining 92.2% of FGP variation in the F2 population. Through further physical mapping, a co-segregate and co-dominant DNA marker with the locus, flo7(t) was successfully developed, by which, thereby, breeding efficiency of rice cultivars having proper dry milling adaptability with high yield potential or useful functional materials would be improved. 'Suweon 542' maintained the early maturity of the wild type, Namil, which can be used in rice-wheat double cropping systems in Korea not only for improved arable land but also for sharing flour production facilities. In addition to the high susceptibility against major rice diseases, nevertheless, another possible drawback of 'Suweon 542' is the high rate of viviparous under prolonged rainfall during the harvesting season. To overcome susceptibility and vivipary of 'Suweon 542', the progeny lines, derived from the crosses 'Suweon 542' and 'Jopyeong', an early maturing rice cultivar with multiple resistance against rice blast, bacterial blight, and rice strip virus, and 'Heugjinju', a anthocyanin pigment containing black rice cultivar, were intensively evaluated. As the outputs, three dry milling suitable rice elite lines, 'Jeonju614', 'Jeonju615', and 'Jeonju616' were developed.

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굴과 상추에서 노로바이러스의 대체모델 feline calicivirus의 효율적 검출법 개발 (Development of Protocol for the Effective Detection of Feline Calicivirus as Norovirus Surrogate in Oyster and Lettuce)

  • 이수연;장금일;우건조;곽효선;김광엽
    • 한국식품과학회지
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    • 제39권1호
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    • pp.71-76
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    • 2007
  • 본 연구에서는 노로바이러스의 대체모델인 feline calicivirus(FCV)를 이용하여 식품 중에 존재하는 식중독 바이러스의 신속검출방법을 개발하였다. 일반적으로 식품 중에 존재하는 식중독 바이러스를 검출하기 위해서는 식품으로부터 바이러스를 분리시키고, 분리된 바이러스를 농축한 뒤 RT-PCR 방법을 이용하여 검출하는데, 각각의 과정을 수행하는 데 있어서 다양한 문제점이 존재하고 있다. 첫째, 식품으로부터 바이러스를 분리하고 농축하는 과정에서 시간이 많이 걸린다는 것과 둘째, 농축하여 추출된 바이러스 RNA로 RT-PCR 방법을 수행하는데 있어서 잔존하는 식품 성분이 PCR 반응을 저해하여 검출효율이 낮아지는 문제점이 있다. 본 연구에서는 0.2 ${\mu}m$ syringe filter를 사용하여 바이러스 분리과정에서 PCR 저해물질인 식품성분을 추가적으로 제거시켰으며, 농축과정에서는 분쇄된 얼음에 방치하는 방법을 사용하여 overnight하는 과정을 3시간으로 단축시켰다. 농축된 바이러스의 RNA 추출물에 잔존하는 PCR 저해물질을 희석함으로서 보다 높은 검출효율을 얻을 수 있었다. 본 연구를 통해 개발된 신속검출법은 굴과 상추에서 노로바이러스의 신속검출을 위한 방법으로 적용이 가능할 것으로 판단되며, 또한 다양한 식품에서 식중독 바이러스의 검출 효율을 향상시키는데 기여할 것으로 생각된다.

로타바이러스 감염성 설사에 대한 항-로타바이러스 난황항체의 치료 효과 (Therapeutic Effect of Anti-Rotavirus Chicken Egg Yolk Immunoglobulin(IgY) on Diarrhea by Infection of Rotavirus)

  • 임인석;이호석;김원용;최응상;정동혁;정후길;윤승섭;전호남
    • Clinical and Experimental Pediatrics
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    • 제48권12호
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    • pp.1354-1361
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    • 2005
  • 목 적 : 로타바이러스는 세계적으로 5세 이하 영유아 설사증의 1/3 이상을 차지하는 주요 원인체이다. 이 로타바이러스에 의한 급성위장관염의 치료는 탈수교정을 위한 경구 및 비경구 수액요법 외엔 특이한 치료법이 없는 상태로서 로타바이러스 감염에 대한 새로운 치료법이 필요한 상태이다. 이에 본 연구에서는 로타바이러스에 대한 난황 항체를 투여하여 환아에서의 로타바이러스 억제 효과 및 임상증상의 치료효과를 조사하였다. 방 법 : 본 연구는 두 부분으로 나누어 첫째 로타바이러스 위장관염 환아의 분변에서 바이러스 입자 감소 정도를 보았으며 둘째 환아에게 난황황체를 환아에게 직접 투여하여 임상증상이 개선되는 지를 보았다. 첫째로, 2001년 12월부터 2002년 9월까지 중앙대학교 용산병원 소아과에 급성 장관염을 주요 증상으로 내원한 환아의 분변을 대상으로 역수동 적혈구 응집반응으로 로타바이러스 항원의 존재 유무를 조사하였고 이중 양성인 13명의 환아의 3일간에 각각 걸친 분변시료를 재료로 정량 PCR을 시행하여 바이러스 양의 감소를 측정하였다. 13명 중 7명에게는 난황항체가 투여되었고 6명에게는 위약이 투여되었다. 둘째로는 같은 기간 로타바이러스 감염으로 진단된 36명을 대상으로 하여 18명에는 항-로타바이러스 난황항체가 함유된 난황 2.5 g을 5 mL의 물에 부유하여 하루 2회(아침, 저녁) 3일 동안 투여하였고 위약군 18명에게는 난황항체가 함유되지 않은 난황을 같은 기간 동안 같은 양을 투여한 이후 양군에서 치료 전후의 발열기간, 설사 횟수, 구토 횟수, 전해질 및 혈청 생화학치, 입원기간, 경구 섭취량, 소변 횟수를 비교 관찰하였다. 결 과 : 첫 번째 실험에서, 정량 PCR을 이용하여 환아 분변에서 바이러스 입자의 감소를 측정한 결과 위약군에서는 지속적으로 증가를 보였으나 투여군에서는 지속적인 바이러스 입자의 감소를 보였다. 두 번째 실험에서는 환아의 임상증상 치료효과 면에서 투여군에서 위약군에 비해 3일째에 설사 증상의 개선을 보였다. 결 론 : 항로타바이러스 난황항체의 로타바이러스 감염에 대한 치료효과를 본 결과 환아의 분변에서 바이러스 입자량의 감소와 함께 설사 증상의 개선의 치료 효과를 보이기 때문에 항로타바이러스 난황항체는 로타바이러스 감염증의 치료제가 될 수 있을 것으로 생각된다.