• IMMUNE NETWORK
• /
• v.3 no.3
• /
• pp.219-226
• /
• 2003

Background: Phage display is the most widely used technique among display methods to produce monoclonal antibody fragment with a specific binding activity. Having a large library for efficient antibody display/selection is quite laborious process to have more than $10^9$ members of transformants. To overcome these limitations, several in vitro selection approaches have been reported. Ribosome display that links phenotypes, proteins, directly to genotype, mRNA, is one of the in vitro display methods. Ribosome display can reach the size of scFv library up to $10^{14}$ molecules and it can be further diversified during PCR steps. To select the high affinity scFv from one pot library, we established ribosome display technique by modifying the previously reported eukaryotic translation system. Methods: To establish the antibody selection system by ribosome display, we used 3D8, anti-DNA antibody. A 3D8 scFv was synthesized in vitro by an in vitro transcription-translation system. The translated 3D8 scFv and the encoding 3D8 mRNA are connected to the ribosome. These scFv-ribosome-mRNA complexes were selected by binding to their specific antigens. The eluted mRNAs from the complexes are reverse transcribed and re-amplified by PCR. To apply this system, antibody library from immunized mouse with terminal protein (TP)-peptide of hepatitis B virus DNA polymerase TP domain was also used. This TP-peptide encompasses the 57~80 amino acid residues of TP. These mRNA/ribosome/scFv complexes by our system were panned three times against TP-peptide. The enrichment of antibody from library was determined by radioimmunoassay. Results: We specifically selected 3D8, anti-DNA antibody, against ssDNA as a model system. The selected 3D8 RNAs sequences from translation complexes were recovered by RT-PCR. By applying this model system, we enriched TP-peptide-specific scFv pools through three cycles of panning from immunized library. Conclusion: We show that our translating ribosome complexes are well maintained and we can enrich the TP-specific scFv pools. This system can be applied to select specific antibody from an antibody library.

• Journal of Life Science
• /
• v.25 no.11
• /
• pp.1324-1330
• /
• 2015

The present study was performed to analyze and review the physical and immune responses to overtraining syndrome in humans. Overreaching refers to the initial phage of overtraining syndrome and has been known as a physical fatigue which is mainly from metabolic imbalance. It has been known that overtraining also results in a loss of adaptability which may lead to an attenuation of exercise performance, sleeping disorder, central fatigue, neurohormonal changes, difficulty recovery to physical stress, and immunological changes. Additionally, overtraining syndrome is characterized by persistent fatigue, poor performance in sport due to the prolonged and strenuous physical training. Also, previous studies reported that endurance athletes experienced a high incidence of URTI during intense training and the post training. And also, high-performance athletes reported that suppression of cell mediated and anti-body mediated immune function. NK cell numbers were also reduced in the period of overtraining syndrome. Major components of prevention and treatment for the overtraining syndrome are screening, education, and detraining. Furthermore, the combination of these prevention and treatment strategies will be much helpful. Therefore, the current review will be helpful for athletes and individuals who are at the risk of overtraining syndrome.

• Proceedings of the Korean Journal of Food and Nutrition Conference
• /
• 2000.05a
• /
• pp.5-7
• /
• 2000

국내에서의 식이섬유의 이용은 대부분 저분자량의 수용성 식이섬유를 이용한 음료형태가 주류를 이루고 있으며, 밀기울, 귀리, 옥수수 껍질 등으로 부터 얻는 불용성 식이섬유는 제과 및 제빵등에 이용되어 오고 있다. 수용성 식이섬유는 혈중 콜레스테롤을 저하시키며, 불용성 식이섬유는 변비예방과 장기능 증진효과가 탁월한 것으로 알려져 있다. 그러므로, 대장암과 변비같은 장질환 환자가 날로 증가하고 있는 상황에서 국내산 미이용 자원으로부터 불용성 식이섬유의 기능성 향상과 활용기술을 개발하는 것은 식품산업적 가치가 크다고 할 수 있다. 또한, 국내에서 소비되고 있는 폴리덱스트로스와 올리고당과 같은 기능성 당류의 대부분이 외국에서 수입하여 국내에 보급되고 있기 때문에 큰 외화지출이 되고 있다. 따라서 본고에서는 국내산 미곡 부산물인 밀기울 식이섬유 세포벽의 불용성 구조를 수용성으로 변화시킬 수 있는 변형기술과 얻어진 수용성 식이섬유 소재의 특성을 소개하고자 한다. 즉, 밀기울 세포벽의 수용화를 위한 가공처리는 압출성형 공정에 의하여 실시하였으며, 수용화가 수반되는 압출성형 조건에 따른 식이섬유 소재의 이화학적 특성과 수용화가 일어난 세포벽 성분의 분해기작에 대한 연구결과를 정리하였다. 그리고, 현행 올리고당과 식이섬유의 산업적 생산공정에서 문제점으로 대두되고 있는 효소처리, 산성하에서의 감압가열, 고점도 원료물질의 이송, 생물공학적 공정제어, 반응시간, 잔여 효소의 불활성화 및 재활용, 제조원가 상승 등과 같은 것을 해결하거나 개선할 수 있는 효율적인 압출반응공정의 조건과 생산된 소재 glucooligosaccharide, fructooligosaccharide, galactooligosaccharide 등의 특성에 대하여 논하여 보기로 한다.경우가 많지 않다. 또한 그 발생 경위나 원인요소에 대해서는 일반에게 공개되지 않고 있어, 우리가 실천하여 식중독을 예방할 수 있는 정보가 함께 제시되지 못하는 형편이다. 따라서 본 고에서는 최근 수년간 외국에서 학교급식을 비롯한 집단급식 등에서 발생한 식중독 사례를 중심으로 고찰, 분석하였다. 이로부터 식중독 발생에 기여한 주요 원인을 찾아내고 여기서 얻어지는 교훈을 토대로 식중독 발생을 예방 및 최소화할 수 있는 실천적 대처 방안을 모색해 보고자 한다.중의 E. coli O157:H7이 연관되어있다는 보고도 있다. 목장 방문시 원유를 마신 어린이에서 HUS가 유발하였고, 균분리 결과, 농장의 소와 어린이에서 유사한 E. coli O157:H7이 분리된 바 있다. 본 연자들은 베로톡신 유형, plasmid profiles, RFLP 분석, phage type을 기초로하여 소에서 분리된 E. coli O157:H7이 사람유래주와 매우 유사함을 확인하였다.mm 및 2-6mm 난포에서의 101개(67.8%) 및 47개(50.0%) 보다 매우 낮게 나타났다. 전체 회수 난포란수도 4등급이 59.1%(149/252)로써 1, 2, 3등급의 0.4% (1/252), 7.6%(19/252) 및 32.9%(83/252)보다 높게 나타났다. 1회 평균 회수 난포란은 $\leq$2mm 난포에서 4.8$\pm$3.7개로써 2-6mm(3.0$\pm$3.4개) 및 $\geq$6mm (0.3$\pm$0.6개)보다 높았으며, 1회당 평균 8.1$\pm$5.1개의 난포란을 회수하였다. investigation can be separated into sa

• Korean Journal of Microbiology
• /
• v.17 no.4
• /
• pp.165-178
• /
• 1979

Phages of Lactobaciilus casei (PLC) isolated from plant drainage were classified and characterized. The results are as follows : 1. On the basis of host range pattern, phages could be divided into 2 groups (PLC-B and PLC-C). PLC-B group phages could be further divided into 5 sub-groups $(B_1, \;B_2, \;B_3, \;B_4, \;and\;B_5)$. Although PLC-C group phages had the same host range, they could be also divided into 2 sub-groups $(C_1\;and\;C_2)$ by morphlogical type. 2. It was $B_3$ group phages that represented a major proportion (44.4%) of phages tested. However, $B_1$ group phages were shown to have the widest host range. 3. Electron micrographs revealed that the phages fell into three different morphological types. $(B_1, \;B_2, \;and\;B_3)$ group phages hd a hexagonal head (52nm in diameter) and a sheathless noncontractile (245 nm in length). $B_4\;and\;C_2$ group phages had a hexagonal head (56 nm) and a short flexible tail (169nm) having no sheath. $B_5\;and\;C_1$ group phages were shown to have a hexagonal head (81 nm) and a contractile tail (140 nm) having a sheath, a base plate and tail fibers. 4. The inactivation of the phages by antisera indicated that serological relationships correlated completely with morphological types. 5. $B_1, \;C_1\;and\;C_2$ group phages produced a large (1, 2 mm in diameter) plaque with a clear ring. The morphology of plaques of $B_3\;and\;B_5$ group phages was the same as those produced by the above, but the average plaque sizes for $B_3\;and\;B_5$ were 0.8 mm abd 0.5 mm, respectively. $B_2\;and\;B_4$ group phages produced a small (0.5 mm) turbid plaque with an irregular edge. 6. The latent period and the average burst size of $B_1\;and\;B_3$ group phages were 90 min and 100, respectively. These phages reuqired calcium ions for their miltiplication. 7. $B_3$ group phages could not be absrobed to R-variant $KC_1$. 8. The order of resistance of phages to heat was $B_2\;>\;B_1, B_4\;and\;B_5\;>\;B_3\;and\;C_2, \;B_5$ group phages were more stable than $B_3$ in various pH values. $C_2$ group phages were more sensitive to UV irradiation than $B_1\;and\;B_3$ group phages. 9. Strains YIT9018 and IAM 1043 were induced by mitomycin C treatment. Phage particles detected in the lysates had a hexagonal head (38 and 49 nm, respectively), but no tail. Any sensitive indicator strain could not be isolated in spite of repaeated trials.

• Journal of Radiopharmaceuticals and Molecular Probes
• /
• v.1 no.1
• /
• pp.38-45
• /
• 2015

Most tumors are believed to overexpress several receptors, and small peptides targeting these receptors were developed for diagnosis and tumor therapy during past decade. Here we report that fibroadenoma can be visualized by bladder cancer specific peptide. A 9-mer bladder cancer specific peptide, which was discovered from the phage display method, was synthesized by peptide synthesizer, and additional tyrosine was conjugated at the N-terminal for radioiodination (Y-BP). Y-BP was radiolabeled with $^{131/124}I$ using Iodogen tube. The rat treated with N-butyl-N-(4-hydroxybutyl)nitrosamine for 8 weeks was allowed to grow until large size tumor was developed under axilla. The tumor model was microPET imaged sequentially using [$^{18}F$]FDG and radioiodinated $^{124}I-Y-BP$. The tumor was excised and examined by immunostaining studies. Radioiodinated $^{124}I-Y-BP$ was purified using fast protein liquid chromatography (FPLC) in > 90% radiochemical purity. The whole tumor was well visualized by [$^{18}F$]FDG with several intense focal uptake within tumor. The tumor was also clearly seen with $^{124}I-Y-BP$ at 4 h post-injection, and to our surprise the tumor uptake of $^{124}I-Y-BP$ lasted up to three days. The tumor was diagnosed histologically as a fibroadenoma derived from mammary gland. In conclusion, the bladder cancer specific peptide showed the good potential as a new radiotracer for the detection of breast fibroadenoma.

• The Journal of the Korean Society for Microbiology
• /
• v.21 no.1
• /
• pp.73-95
• /
• 1986

A total of 5,462 isolates suspicious of Salmonella, Shigella and E. coli which were isolated during 1983 to 1985 by 12 City Hygine Laboratories and General Hospital Laboratories were received and identified at the National Salmonella Center, Seoul, Korea. The result of identification of these strains were summarized as follows: 1. It was confirmed that the total organisms broke down into 2,014 strains of Salmonella 1,294 of which were S. typhi, 887 strains of Shigella and 2,561 strains of E. coli. 2. For seasonal distribution of enteric pathogens, July was the month with the highest out breaks of salmonellosis, May was the month of Shigellosis, and April was of the highest month it in the case of E. coli. 3. Salmonella typhi with the highest incidence of isolation was shown to belong to various phage types, especially with the strains detected in Seoul. M1 type was widely distributed all over the country, but the majority was E1 type in 1983. 4. For age distribution of patients, the 20-29 age group had the highest incidence of salmonellosis whileas the 1 to 9 age group had the highest incidence of Shigellosis. 5. For sexuly distribution of Salmonella and Shigella infections seemed to be relatively higher in the female than in the male. However, E. coli. had no relationship to both sex. 6. The antibiotic sensitivity patterns of S. typhi cultures showed a tendancy to be resistant to colistin, gentamycin, neomycin, tetracycline and streptomycin. 7. The isolates of S. paratyphi-A, S. typhimurium and S. enteritidis seemed to have a tendency of multiple drug resistance. 8. 93.9 percent of 1,568 E. coli strains showed negative reactions to the antisera of enteropathogenic E. coli and 15.6 percent of them produced a heat-labile enterotoxin, but positive reaction to the antisera was 6.1 percent and 11.6 percent of them producled the enterotoxin.

• Journal of Life Science
• /
• v.17 no.2 s.82
• /
• pp.272-278
• /
• 2007

Proteases are enzymes that break peptide bonds between amino acids of other proteins and occupy a crucial position with respect to their applications in both physiological and commercial fields. In order to screen new source of protease, bacteria producing extracellular proteases at low temperature were isolated from deep sea water of East Sea, Korea. A bacterium showing the best growth rate and production of an extracellular protease at low temperature was designated HJ 47. The DNA sequence analysis of the 16S rRNA gene, phenotypic tests and morphology led to the placement of this organism in the genus Pseudoalteromonas. Although maximal growth was observed at $37^{\circ}C$, enzyme production per culture time was maximum at $20^{\circ}C$. At this temperature, extracellluar protease production was detected from the end of the exponential phage to stationary phase, and maximal at 15 hours after initial production. The optimum temperature and pH of the protease were found to be $35^{\circ}C$ and 8.

• Journal of Ginseng Research
• /
• v.41 no.2
• /
• pp.134-143
• /
• 2017

Background: The prevalence of allergic inflammatory diseases such as atopic dermatitis (AD), asthma, and allergic rhinitis worldwide has increased and complete recovery is difficult. Korean Red Ginseng, which is the heat-processed root of Panax ginseng Meyer, is widely and frequently used as a traditional medicine in East Asia. In this study, we investigated whether Korean Red Ginseng water extract (RGE) regulates the expression of proinflammatory cytokines and chemokines via the mitogen-activated protein kinases (MAPKs)/nuclear factor kappa B ($NF-{\kappa}B$) pathway in allergic inflammation. Methods: Compound 48/80-induced anaphylactic shock and 1-fluoro-2,4-dinitrobenzene (DNFB)-induced AD-like skin lesion mice models were used to investigate the antiallergic effects of RGE. Human keratinocytes (HaCaT cells) and human mast cells (HMC-1) were also used to clarify the effects of RGE on the expression of proinflammatory cytokines and chemokines. Results: Anaphylactic shock and DNFB-induced AD-like skin lesions were attenuated by RGE administration through reduction of serum immunoglobulin E (IgE) and interleukin (IL)-6 levels in mouse models. RGE also reduced the production of proinflammatory cytokines including $IL-1{\beta}$, IL-6, and IL-8, and expression of chemokines such as IL-8, thymus and activation-regulated chemokine (TARC), and macrophage-derived chemokine (MDC) in HaCaT cells. Additionally, RGE decreased the release of tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), $IL-1{\beta}$, IL-6, and IL-8 as well as expressions of chemokines including macro-phage inflammatory protein $(MIP)-1{\alpha}$, $MIP-1{\beta}$, regulated on activation, normal T cell expressed and secreted (RANTES), monocyte chemoattractant protein (MCP)-1, and IL-8 in HMC-1 cells. Furthermore, our data demonstrated that these inhibitory effects occurred through blockage of the MAPK and $NF-{\kappa}B$ pathway. Conclusion: RGE may be a useful therapeutic agent for the treatment of allergic inflammatory diseases such as AD-like dermatitis.

• Proceedings of the Korean Journal of Food and Nutrition Conference
• /
• 1999.06a
• /
• pp.39-57
• /
• 1999

Chitin은 새우나 게 등을 비롯한 해산 갑각류의 가공 폐기물로 버려지는 부패, 악취발생 등의 환경공해를 일으키며 처리비용 등 여러 가지 문제를 야기하던 물질이지만 최근에는 부가가치가 높은 chitosan 및 chitooligosaccarides로 가공되기 때문에 폐기물의 자원화와 미이용 자원의 개발 등 여러 가지 측면에서 기대가 모아지고 있는 다당류이다. 더욱이 chitin의 탈아세틸화로 얻어지는 chitosan은 polycation의 성질을 가지고 있어서 현재 응집제로서 각종 공업 분야에 널리 이용되고 있으며 최근에는 그 외에도 다양한 생리적 기능성을 가지고 있다는 사실이 밝혀지면서 의료, 생호학, 화장품, 식품, 화학 공업 및 농업용 등 다양한 분야에 응용 가능한 신소재로서 기대가 높아지고 있다. 그런, chitosan은 생리적 기능은 우수하지만 물에 용해되지 않고 점도가 높으며 떫은맛이 나기 때문에 식품을 비롯한 기타 응용분야에 이용이 제한되고 있다. 따라서 많은 연구자들은 이와 같은 물리적인 장애를 개선함으로써 chitosan 자체가 갖는 우수한 생리적 기능을 이용할 뿐만 아니라 항종양성을 비롯한 새로운 생리적 기능을 갖는 chitosan 유도체의 개발을 시도하고 있다. Chitosan 유도체 중에서도 chitooligosaccharides는 chitosan의 가수분해로 얻어지는 저분자 화합물이므로 물에 잘 용해도며 점도가 낮고 용액이 단맛을 낼뿐만 아니라 chitosan이 갖지 않는 새로운 생리적 기능성을 갖는 것으로 알려지면서 관심이 집중되기 시작하였다. 본고에서는 chitosan과 chitooligosaccarides의 생리 활성 기능에 대한 지금까지의 연구결과를 소개하고자 한다.에 대한 면밀한 사안이 밝혀지는 경우가 많지 않다. 또한 그 발생 경위나 원인요소에 대해서는 일반에게 공개되지 않고 있어, 우리가 실천하여 식중독을 예방할 수 있는 정보가 함께 제시되지 못하는 형편이다. 따라서 본 고에서는 최근 수년간 외국에서 학교급식을 비롯한 집단급식 등에서 발생한 식중독 사례를 중심으로 고찰, 분석하였다. 이로부터 식중독 발생에 기여한 주요 원인을 찾아내고 여기서 얻어지는 교훈을 토대로 식중독 발생을 예방 및 최소화할 수 있는 실천적 대처 방안을 모색해 보고자 한다.중의 E. coli O157:H7이 연관되어있다는 보고도 있다. 목장 방문시 원유를 마신 어린이에서 HUS가 유발하였고, 균분리 결과, 농장의 소와 어린이에서 유사한 E. coli O157:H7이 분리된 바 있다. 본 연자들은 베로톡신 유형, plasmid profiles, RFLP 분석, phage type을 기초로하여 소에서 분리된 E. coli O157:H7이 사람유래주와 매우 유사함을 확인하였다.mm 및 2-6mm 난포에서의 101개(67.8%) 및 47개(50.0%) 보다 매우 낮게 나타났다. 전체 회수 난포란수도 4등급이 59.1%(149/252)로써 1, 2, 3등급의 0.4% (1/252), 7.6%(19/252) 및 32.9%(83/252)보다 높게 나타났다. 1회 평균 회수 난포란은 $\leq$2mm 난포에서 4.8$\pm$3.7개로써 2-6mm(3.0$\pm$3.4개) 및 $\geq$6mm (0.3$\pm$0.6개)보다 높았으며, 1회당 평균 8.1$\pm$5.1개의 난포란을 회수하였다. in

• Biomedical Science Letters
• /
• v.5 no.2
• /
• pp.135-145
• /
• 1999

A total of 45 Staphylococcus aureus strains from clinical samples were tested for the biochemical test and antibiotic susceptibility test. Forty-five S. aureus strains were subjected to the molecular epidemiological study by susceptiblity test, antibiogram, bacteriophage typing, polymerase chain reaction and mec-associated hypervariable region gene in order to detect of mecA gene which was one of the structural gene related to antibiotic resistant expression factors. Three of 15 mecA-negative S. aureus isolates were classified as oxacillin resistant despite borderline minimal inhibitory concentration values. Methicillin susceptiblities were completely consistent with PCR results for these strains. On the other hand, 4 of 30 mecA-positive isolates yielded results in the oxacillin and methicillin susceptibility tests which were discrepant from those of PCR analysis. Except for SA6, the methicillin resistant S. aureus strains tested were highly resistant to penicillin, oxacillin, gentamicin, and chloramphenicol. In the phage typing, 27 strains were typable. The Iytic group III was as many as 12 strains, and 7 of 12 were 75/83A/84 type. In the PCR of specific mecA gene probe with chromosomal DNA of 30 methicillin resistant S. aureus, the amplified DNA band of 533 bp was confirmed in 30 strains and not in methicillin sensitive S. aureus. The single amplified band of hypervariable region related to mec was investigated in all of 30 methicillin resistant S. aureus, but in methicillin sensitive S. aureus it was amplified. The size of PCR products was between 200 bp and 600 Up. Four units was directly repeated.