• Title/Summary/Keyword: vaccine preparation

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Surface Mmodification of Poly(DL-lactide-co-glycolide) Nanoparticle (Poly(DL-lactide-co-glycolide) 나노입자의 표면 수식)

  • Oh, Yu-Mi;Jung, Taek-Kyu;Chi, Sang-Cheol;Shin, Byung-Cheol
    • Journal of the Korean Chemical Society
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    • v.47 no.6
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    • pp.601-607
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    • 2003
  • We studied on preparation of nanoparticles modified surface using biodegradable polymer, poly(DL-lactide-co-glycolide) (PLGA). Two kinds of PLGA nanoparticles were prepared by a spontaneous emulsification solvent diffusion (SESD) method using cetyltrimethylammonium chloride (CTAC) and tetradecyltrimethylammonium bromide (TTAB) as a cationic surfactant and polyethylene glycol-block-polypropylene glycol copolymer (Lutrol F68) as a nonionic surfactant. Model protein was coated on the surface of nanoparticles by the ionic complexation. The model protein was that influenza vaccine ($H_3N_2,\;H_1N_1$, B strain) labeled with NHS-fluorescein. The sizes of cationic nanoparticles were 140-160 nm and the surface charges were 50-60 mV. The sizes of nonionic nanoprticles were 80-90 nm and the surface charge was -10 mV. After coating vaccine on the surface of nanoparticles, the sizes of cationic nanoparticles were increased to 380-400 nm and the size of nonionic nanoparticles was not increased. The amount of coated vaccine on the cationic nanoparticles was 22.73 ${\mu}g$/mg.

Leading Study for the Restoration of Pediatric Immunization Systems in North Korea following Reunification of Korean Peninsula (통일 후 북한지역 영·유아 대상 예방접종체계 수복을 위한 선행연구)

  • Song, Seulki;Bang, Joon Seok
    • Korean Journal of Clinical Pharmacy
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    • v.25 no.4
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    • pp.223-230
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    • 2015
  • Background: 8,000 children in North Korea died before they reached the 1 year after births in 2013. The high mortality rate of children under five years of age is mainly caused by infectious diseases and malnutrition. The need for national pediatric immunization and supply of vaccines will be vital when the abrupt reunification occurs. Objective: The purpose of this study is to scrutinize the pediatric immunization coverage of North Korea. Additionally it is to estimate the amount and the costs needed to vaccinate. Methods: The target population is the children of North Korea. The method is based on a pre-survey and an interview of North Korea defectors. The target interviewees searched for are as follows; doctors, teachers, and others. The interview includes questions on mortality rates and immunization coverage. The analysis is largely based on the statements of the health care providers within the selected group. Results: The interviewees are 8; 7 female and 1 male. The birth years range from 1956 to 1982. 3 out of 8 are former health care providers; a doctor, a pharmacist, and a nurse. The morbidity rate of infectious diseases exceeds the data from WHO. The immunization coverage is nearly 0% after 1980s. In order to ensure the welfare of North Korean children, at least 8,234,000 vaccine doses, requiring over 105 million U.S. dollars, are needed. Conclusion: The morbidity rate of infectious disease in North Korea is conspicuous. The preparation for supply and expenditure of vaccines is vital.

Immunogenicity and Protective Efficacy of Solubilized Merozoite-enriched Theileria sergenti Immunogens I: Protection against Homologous Stabilate Challenge (Theileriu sergenti merozoite 수용성 항원의 항원성과 면역성)

  • 백병걸;최인혁
    • Parasites, Hosts and Diseases
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    • v.30 no.2
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    • pp.133-140
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    • 1992
  • Theileria sergenti were isolated from infected erythrocytes by hypotonic Iysis, and soluble meroBoite antigens were purified by sonication and differential centrifugation. The preparation contained 29, 34, 35 and 105 kD immuno-dominant polypeptides. The soluble antigens (0.5 mg/ml) were prepared and fortified with Freund's adjuvant. Five month old naive Korean calves were subcutaneously inoculated with the preparation and a booster dose was administered 4 weeks later Nine weeks after the booster dose, vaccinates and controls were challenged with a homologous stabilate (5.6×106 RBC/dose, 40% Parasitemia). All animals were monitored for hematocrit, total erythrocyte count, parasitemia and for the specific antibody by Western immuno- blot (WB) and indirect immuno-auorescent antibody(IFA) test. By 18 weeks after vaccination (6 weeks after the challenge), vaccinated cattle had an average IFA titer of 1 : 10,240 compared with 1 : 1,280 of the controls. The vaccinates showed ne91igib1e change in hematocrit and total RBC count whereas control animals showed significant (P<0.05) hematological chanties and associated anemia. After vaccination and challenge, the antibody responses demonstrated that vaccination had induced significant production of antibody to the 29 and 35 kD polypeptides. The latter polypeptide was much more strongly recognized by the vaccinated animals, and thus it may be a potential candidate for the vaccine.

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The effects of adjuvants and vaccine against edwardsiellosis in tilapia, Oreochromis nioticus (틸라피아의 에드와드병에 대한 백신과 Adjuvant의 효과)

  • Lee, Joo-Seok;Park, Soo-Il
    • Journal of fish pathology
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    • v.5 no.1
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    • pp.19-27
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    • 1992
  • A formalized bacterin(FKC) of Edwardsiella tarda, conjugated with/without Freund's complete adjuvant(FCA), Freund's incomplete adjuvant(FIA) and potassium aluminium sulfate(PAS), was administered by intraperitoneal injection to tilapia respectively. The tilapia were reared under indoor recirculating filter system. There were agglutinin titer increment in the tilapia which were administered not only FKC bacterin but also adjuvanted vaccines. But the producing time of the highest level of antibody in the immunized tilapia was different among vaccine preparation, i. e. FKC+FCA group and PAS+FKC group are between the 2nd and the 4th week, and FKC alone group is the 2nd week. In the challenge experiment with $2.5{\times}10^7\;CFU/ml$ or $2.5{\times}10^8\;CFU/ml$ of E. tarda T1123 to the tilapia immunized with vaccines, RPS above sixty was recorded both challenge dose in the tilapia which were 3 weeks after immunization with FKC+FCA, PAS+FKC and FKC alone, and the former dose 8 weeks after immunization with FCA+FKC and PAS+FKC. There were some resistance enhancement against E. tarda in the tilapia which were injected with adjuvant alone than that of control. As the results, the FCA or PAS adjuvanted vaccine is effective to sustain the defensible period against edwardsiellosis.

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Various Types and Manufacturing Techniques of Nano and Micro Capsules for Nanofood

  • Kim, Dong-Myong
    • Journal of Dairy Science and Biotechnology
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    • v.24 no.1
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    • pp.53-63
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    • 2006
  • Nano and micro capsulation (NM capsulation) involve the incorporation for nanofood materials, enzymes, cells or other materials in small capsules. Since Kim D. M. (2001) showed that a new type of food called firstly the name of nanofood, which means nanotechnology for food, and the encapsulated materials can be protected from moisture, heat or other extreme conditions, thus enhancing their stability and maintaining viability applications for this nanofood technique have increased in the food. NM capsules for nanofood is also utilized to mask odours or tastes. Various techniques are employed to form the capsules, including spray drying, spray chilling or spray cooling, extrusion coating, fluidized bed coating, liposome entrapment, coacervation, inclusion complexation, centrifugal extrusion and rotational suspension separation. Each of these techniques is discussed in this review. A wide variety of nanofood is NM capsulated - flavouring agents, acids, bases, artificial sweeteners, colourants, preservatives, leavening agents, antioxidants, agents with undesirable flavours, odours and nutrients, among others. The use of NM capsulation for sweeteners such as aspartame and flavors in chewing gum is well known. Fats, starches, dextrins, alginates, protein and lipid materials can be employed as encapsulating materials. Various methods exist to release the ingredients from the capsules. Release can be site-specific, stage-specific or signaled by changes in pH, temperature, irradiation or osmotic shock. NM capsulation for the nanofood, the most common method is by solvent-activated release. The addition of water to dry beverages or cake mixes is an example. Liposomes have been applied in cheese-making, and its use in the preparation of nanofood emulsions such as spreads, margarine and mayonnaise is a developing area. Most recent developments include the NM capsulation for nanofood in the areas of controlled release, carrier materials, preparation methods and sweetener immobilization. New markets are being developed and current research is underway to reduce the high production costs and lack of food-grade materials.

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Recombination and Expression of eaeA Gene in Enterohemorrhagic Escherichia coli O157:H7

  • Kim, Hong;Kim, Jong-Bae
    • Biomedical Science Letters
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    • v.8 no.3
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    • pp.107-113
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    • 2002
  • Enterohemorrhagic Escherichia coli (EHEC) strains of serotype O157:H7 have been shown to colonize the intestinal epithelial cell by the attaching and effacing (AE) mechanism. The AE lesion is mediated by an intimin, of which production and expression are controlled by a 3-Kb eaeA gene located EHEC chromosomal DNA. If the eaeA gene is mutated, EHEC O157:H7 strains lose capacity of adhesion to intestinal epithelial cells. In this study, a 891 bp of the 3'-end region of a gamma intimin was amplified by polymerase chain reaction (PCR). The PCR product was inserted into pSTBlue-1 cloning vector and transformed into DE3 (BL21) competent cell. After plasmid mini-preparation and restriction enzyme digestion of eaeA/891-pSTBlue-1 vector, target eaeA gene was re-inserted into pET-28a expression vector and was transformed. Then the expression of recombinant eaeA/891 (891 bp) gene was induced by isopropyl-$\beta$-D-thiogalactopyranoside (IPTG). The expression of the 40-KDa recombinant protein was identified in SDS-PAGE and confirmed by immunoblotting using the His.Tag$^{\circledR}$ and T$_{7}$.Tag$^{\circledR}$ monoclonal antibody. This recombinant protein expressed by eaeA gene could be applied in further studies on the mechanisms of E. coli O157:H7 infection and the development of recombinant vaccine.

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SWATH-based Comparative Proteomic Analysis of the Mycobacterium bovis BCG-Korea Strain

  • Lee, Won-Kyu;Baek, Je-Hyun;Ryoo, Sung Weon;Yu, Yeon Gyu
    • Bulletin of the Korean Chemical Society
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    • v.35 no.3
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    • pp.933-937
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    • 2014
  • A derivative of Mycobacterium bovis Bacillus Calmette-Guerin (BCG) has been used for the preparation of tuberculosis vaccines. To establish a Korean tuberculosis vaccine derived from BCG-Pasteur $1173P_2$, genome sequencing of a BCG-Korea strain was completed by Joung and coworkers. A comparison analysis of the genome sequences of the BCG-Pasteur $1173P_2$ and BCG-Korea strains showed marginal increases in the total genome length (~0.05%) and the number of genes (~4%) in the BCG-Korea genome. However, how the genomic changes affect the BCG-Korea protein expression levels remains unknown. Here, we provide evidence of the proteomic alterations in the BCG-Korea strain by using a SWATH-based mass spectrometric approach (Sequential Window Acquisition of all THeoretical mass spectra). Twenty BCG proteins were selected by top-rank identification in the BCG proteome analysis and the proteins were quantified by the SWATH method. Thirteen of 20 proteins showing significant changes were enough to discriminate between the two BCG proteomes. The SWATH method is very straightforward and provides a promising approach owing to its strong reliability and reproducibility during the proteomic analysis.

Expression and diagnostic application of p12 protein of African swine fever virus by recombinant baculovirus (재조합 baculovirus에 의한 아프리카 돼지콜레라바이러스 p12 단백질의 발현과 진단적 적용)

  • Choi, Kang-Seuk;Choi, Cheong-up;Kim, Yong-Joo
    • Korean Journal of Veterinary Research
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    • v.45 no.1
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    • pp.63-70
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    • 2005
  • African swine fever (ASF) is an infectious disease of domestic and wild pigs for which there is no vaccine in the world. A proper surveillance of viral activity and a timely response to ASF outbreaks depend upon the rapid diagnosis of ASF viral infection. Internationally prescribed enzyme-linked immunosorbent assay (ELISA) is a fast, sensitive test routinely used in the diagnosis of the ASF. However, inactivated whole ASF virus antigen used in this test is a tedious to prepare and has a risk of outside exposure of infectious virus by laboratory accident during the preparation. An ASF virus noninfectious recombinant antigen is a safe and easily produced alternative antigen for use in diagnostic assay. We have cloned the ORF O61R gene of the ASF virus to generate a recombinant baculovirus producing the p12 protein in insect cells under control of the polyhedrin promoter as non-fusion protein. When used in an indirect ELISA, the p12 antigen showed reactivity with all known ASF positive pig sera but not with negative pig sera. Our results indicated that the p12 protein would be one of alternative antigens for diagnosis of the ASF.

Immune Responses of BALB/c Mice Administrated via Oral Route to a Combined Salmonella Typhimurium Ghost Vaccine (복합 살모넬라 타이피무리움 고스트 백신의 마우스 구강 투여에 의한 면역 응답)

  • Kim, Pan Gil;Ha, Yeon Jo;Lee, Su Man;Kim, Sam Woong;Gal, Sang Wan
    • Journal of Life Science
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    • v.25 no.11
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    • pp.1197-1203
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    • 2015
  • Salmonella Typhimurium (ST) JOL389 and χ3339 are strong virulent strains against mouse. ST χ8554 is derived by deletion of the asd gene from ST χ3339. Plasmid pMMP184 carrying a ghost cassette was transformed into ST χ8554, and ST χ8554 ghost cells were prepared and administrated via the oral route to BALB/c mice. Change in the amount of total IgG was not elicited to boosting of single ST χ8554 ghost cells, but the content was increased from 6 weeks after the 3rd administration. However, when the ST JOL389 ghost cells is administered together with ST χ8554 ghost cells, the content of total IgG was increased in 2 weeks post primary administration. It was found that the content of total IgG of the group mixed with ST JOL389 ghost cells showed an increased value of 8 times or more at 10 weeks when compared with the group of ST χ8554 ghost cells. The content of IgG1, IgG2a, and sIgA in both groups increased from 4 weeks postprimary administration. As a challenge test of virulent ST χ3339, χ8554 (pMMP184) and χ8554 (pMMP184)/JOL389 ghost cell groups showed protection of 50% or more when compared to the control group. These results suggest that the preparation of combined ghost cells from a strong virulent ST increases immunity more than a single strain.

Genome characterization and mutation analysis of human influenza A virus in Thailand

  • Rattanaburi, Somruthai;Sawaswong, Vorthon;Nimsamer, Pattaraporn;Mayuramart, Oraphan;Sivapornnukul, Pavaret;Khamwut, Ariya;Chanchaem, Prangwalai;Kongnomnan, Kritsada;Suntronwong, Nungruthai;Poovorawan, Yong;Payungporn, Sunchai
    • Genomics & Informatics
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    • v.20 no.2
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    • pp.21.1-21.14
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    • 2022
  • The influenza A viruses have high mutation rates and cause a serious health problem worldwide. Therefore, this study focused on genome characterization of the viruses isolated from Thai patients based on the next-generation sequencing technology. The nasal swabs were collected from patients with influenza-like illness in Thailand during 2017-2018. Then, the influenza A viruses were detected by reverse transcription-quantitative polymerase chain reaction and isolated by MDCK cells. The viral genomes were amplified and sequenced by Illumina MiSeq platform. Whole genome sequences were used for characterization, phylogenetic construction, mutation analysis and nucleotide diversity of the viruses. The result revealed that 90 samples were positive for the viruses including 44 of A/H1N1 and 46 of A/H3N2. Among these, 43 samples were successfully isolated and then the viral genomes of 25 samples were completely amplified. Finally, 17 whole genomes of the viruses (A/H1N1, n=12 and A/H3N2, n=5) were successfully sequenced with an average of 232,578 mapped reads and 1,720 genome coverage per sample. Phylogenetic analysis demonstrated that the A/H1N1 viruses were distinguishable from the recommended vaccine strains. However, the A/H3N2 viruses from this study were closely related to the recommended vaccine strains. The nonsynonymous mutations were found in all genes of both viruses, especially in hemagglutinin (HA) and neuraminidase (NA) genes. The nucleotide diversity analysis revealed negative selection in the PB1, PA, HA, and NA genes of the A/H1N1 viruses. High-throughput data in this study allow for genetic characterization of circulating influenza viruses which would be crucial for preparation against pandemic and epidemic outbreaks in the future.