HHD Mice를 이용한 대장암세포유래 펩타이드 특이적 CD8⁺ T 세포의 입양전이

정헌순,안인숙,도형기,도명술,Jung, Hun-Soon,Ahn, In-Sook,Do, Hyung-Ki,Lemonnier, Francois A.,Tirosh, Boaz,Tzehoval, Esther,Vadai, Ezra,Eisenbach, Lea,Do, Myoung-Sool 대한면역학회 2004 Immune Network Vol.4 No.1

Background: 1-8D gene is a member of human 1-8 interferon inducible gene family and is shown to be overexpressed in fresh colon cancer tissues. Three peptides 1-6, 3-5 and 3-7 derived from 1-8D gene were shown to have immunogenicity against colon cancer. Methods: To study tumor immunotherapy of these peptides we established an adoptive transfer model. $D^{b-/-}{\times}{\beta}2$ microglobulin (${\beta}2m$) null mice transgenic for a chimeric HLA-A2.1/$D^b-{\beta}2m$ single chain (HHD mice) were immunized with irradiated peptide-loaded RMA-S/HHD/B7.1 transfectants. Spleens were removed after last immunization, and splenocytes were re-stimulated in vitro. Lymphocytes from vaccinated HHD mice were transferred together with IL-2 to the tumor bearing nude mice that were challenged S.C. with the HCT/HHD/B7 colon carcinoma cell line that was found to grow in these mice. Results: Peptide 3-5 was found to be highly effective in CTL activity. Adoptively transferred anti-peptide 3-5 cytolytic T lymphocytes caused significant retardation in tumor growth. Conclusion: This study shows that peptide 3-5 can be the most effective candidate for the vaccine of adoptive immunotherapy against colon cancer.

Competitive Indirect ELISA를 이용한 Bovine Myosin의 동결 변성도 측정

김성배(Seong-Bae Kim),이주운(Ju-Woon Lee),박종흠(Jong-Heum Park),도형기(Hyung-Ki Do),현창기(Chang-Kee Hyun),신현길(Heuyn-Kil Shin) 한국식품과학회 1998 한국식품과학회지 Vol.30 No.4

Bacterial Community Structure Shift Driven by Salinity

Bo Ram Beck(백보람),Wilhelm Holzapfel,Cher Won Hwang(황철원),Hyung Ki Do(도형기) 한국생명과학회 2013 생명과학회지 Vol.23 No.3

강의 하류지역에서 미생물의 군집이 점진적인 염도의 증가에 따라 변한다는 것을 실험적으로 보기 위하여, 경북 형산강의 하류에서부터 연안해역으로 유입되는 곳까지 약 2.91 km 간격으로 0.02%, 1.48%, 2.63%, 3.62%의 염분을 포함하는 물 시료를 얻어 denaturing gradient gel electrophoresis (DGGE)를 수행하였다. 계통분석, 계통수 및 각 시료간의 연관성을 조사한 결과, 미생물 군집의 변화가 염분의 증가에 따라 점진적으로 변화하는 것을 확인하였으며, 이는 염분의 농도가 미생물 군집에 큰 영향을 미치는 요소임을 제시한다. 덧붙여 연안 지역이나 다른 수계환경에 비해 하류지역은 염분의 점진적인 변화로 인해 좁은 면적에 비하여 미생물 다양성이 크고, 이는 곧 특이하고 새로운 종을 찾기에 좋은 장소임을 시사하였다. The influence of a gradual increase in salinity on the diversity of aquatic bacterial in rivers was demonstrated. The denaturing gradient gel electrophoresis (DGGE) was used to analyze the bacterial community shift downstream in the Hyeongsan River until it joins the open ocean. Four water samples were taken from the river showing the salinity gradients of 0.02%, 1.48%, 2.63%, and 3.62%. The samples were collected from four arbitrary stations in 2.91 km intervals on average, and a DGGE analysis was performed. Based on the results of this analysis, phylogenetic similarity identification, tree analysis, and a comparison of each station were performed. The results strongly suggested that the response of the bacterial community response was concomitant to gradual changes in salinity, which implies that salt concentration is a major factor in shifting the microbiota in aquatic habitats. The results also imply a huge diversity in a relatively small area upstream from the river mouth, compared to that in open oceans or coastal regions. Therefore, areas downstream towards a river mouth or delta are could be good starting points in the search for new bacterial species and strains (“biotypes”).

해파리 분말의 상토 첨가물로서의 효과 및 상토의 미생물 군집 변화에 대한 연구

백보람(Bo-Ram Beck),최재호(Jae-Ho Choi),김영록(Young-Rok Kim),차하은(Ha-Eun Cha),도형기(Hyung-Ki Do),황철원(Cher-Won Hwang) 한국토양비료학회 2012 한국토양비료학회지 Vol.45 No.2

해파리 분말을 추가함으로써 나타나는 화분 내의 수분흡착력 및 유지력은 일반 상토보다 월등하고 이는 작물을 관리함에 있어 안정성과 건기 또는 단수로 인한 수분고갈을 예방할 수 있을 것으로 생각된다. 미생물의 경우 DGGE pattern과 염기서열 분석을 통해 확연한 군집변화를 유도하였으며, 식물과 공생관계에서 중요한 Bacteroidetes들의 다양성을 증가시키는 효과를 보았다. 이러한 긍정적인 면이 있지만, 해파리를 분말화하는 공정에 있어서 Potassium alum을 사용 한다는 것이 문제점으로 나타났다. 이 문제를 해결해야만 상품으로써의 가치가 더욱 높아지는데, 이는 해파리가 상토 첨가제로써의 역할도 충분히 할 수 있기 위해서는 phosphate가 inorganic phosphate으로 변환되는 것을 방지해야 하기 때문이다. 요약하자면, 본 실험을 통해 해파리의 잠재적인 자원력 및 고부가 가치화의 가능성은 충분히 나타났다고 생각한다. 이것이 가지는 의미는 식용으로 사용되는 해파리는 제한적이며 독성 해파리들은 거의 사용되지 않기 때문에, 산업적 측면에서 가용성 자원이 늘어난다는 것이다. 또한 환경개선적 측면에서, 독성 해파리률 포함한 해파리들은 여름철 (약 7~8월경)에 해수의 온도 상승과 관련하여 개체수가 급증하여 각 종 어장과 해수욕장에 피해를 주게 된다. 그렇기에 고부가 가치화가 이루어진다면 해파리의 출몰 시기와 이동경로를 예측하여 독성 및 식용불가 해파리들을 사전에 어획함으로써 해양관련 산업에 피해를 줄이고 어민들의 어획종이 늘어남과 동시에 해파리를 이용한 상품을 개발하는 데에 필요한 원자재 확보에도 도움이 될 것이라 생각한다. Recently, the population of toxic and/or unusable jellyfish is increasing during summer along the east coast of Korea, causing massive economical and ecological damage to fisheries, nuclear power plant and marine environment. To solve this problem, this study was carried out using jellyfish as a potential soil additive for horticulture. The jellyfish was solidified and homogenized, then mixed with a commercial bed soil. Allium tuberosum ROTH was planted to control bed soil (BS) and jellyfish powder mixed bed soil groups (Mixed bed soil, MBS), and following parameters were measured during five weeks: water content, electrical conductivity and growth of leaves. At the end of the experiment, bacterial community structures of each pot were analyzed by DGGE. The relative water adsorption of jellyfish powder was about 2.5 times greater compared to its dry weight. The water content of MBS group was significantly higher than BS group 6.5 to 14.2%, and the electric conductivity of MBS group was measured around 2.8 dS/m where BS group was resulted average of 1.8 dS/m. However, the leaves of BS group were grown 30% longer compared to MBS group. DGGE analysis of MBS group was shown in high number of phylum Bacteroidetes and increased diversity of Sphingobacteriia compared to BS group. Jellyfish powder as a soil additive surely will be a good candidate as humectant and microbiota stimulator, although there are several obstacles such as high electrical conductivity and residual alum salt which used for solidification of jellyfish.

천연물 성분을 이용한 환경 유해미생물의 biofilm 생성 저해능 비교에 관한 연구

강은진 ( Eun-jin Kang ),박지헌 ( Ji Hun Park ),진슬 ( Seul Jin ),김영록 ( Young-rok Kim ),도형기 ( Hyung-ki Do ),양웅석 ( Woong-suk Yang ),이재용 ( Jae-yong Lee ),황철원 ( Cher-won Hwang ) 한국환경과학회 2019 한국환경과학회지 Vol.28 No.2

In this study, we investigated the in vitro anti-biofilm activities of plant extracts of chives (Allium tuberosum), garlic (Allium sativum), and radish (Raphanus sativus L.) against environment harmful bacteria (gram-positive Staphylococcus aureus and, gram-negative Salmonella typhimurium and Escherichia coli O157:H7). In the paper disc assay, garlic extracts exhibited the highest anti-biofilm activity. The Minimal Inhibitory Concentration (MIC) of all plant extracts was generally higher for gram-negative bacteria than it was for gram-positive bacteria. Gram-negative bacteria were more resistant to plant extracts. The tetrazolium dye (XTT) assay revealed that, each plant extract exhibited a different anti-biofilm activity at the MIC value depending on the pathogen involved. Among the plant extracts tested, garlic extracts (fresh juice and powder) effectively reduced the metabolic activity of the cells of food-poisoning bacteria in biofilms. These anti-biofilm activities were consistent with the results obtained through light microscopic observation. Though the garlic extract reduced biofilm formation for all pathogens tested, to elucidate whether this reduction was due to antimicrobial effects or anti-biofilm effects, we counted the colony forming units of pathogens in the presence of the garlic extract and a control antimicrobial drug. The garlic extract inhibited the E. coli O157:H7 biofilm effectively compared to the control antimicrobial drug ciprofloxacin; however, it did not inhibit S. aureus biofilm significantly compared to ciprofloxacin. In conclusion, garlic extracts could be used as natural food preservatives to prevent the growth of foodborne pathogens and elongater the shelf life of processed foods.

Production of γ-amino Butyric Acid by Lactic Acid Bacteria in Skim Milk

Jin Young Cha(차진영),Young Rok Kim(김영록),Bo Ram Beck(백보람),Ji Hun Park(박지헌),Cher Won Hwang(황철원),Hyung Ki Do(도형기) 한국생명과학회 2018 생명과학회지 Vol.28 No.2

동해안 지역 수산발효식품과 수산물로부터 다양한 종류의 유산균들을 분리하여 감마아미노낙산(GABA) 활성을 위해 분석을 하였다. 박층크로마토그래피(TLC)를 이용하여 GABA를 생성하는 4개의 균주를 확보하였으며, 16S rRNA sequencing 분석 결과를 통해 FSFL0004, FSFL0005, FSFL0036 균주는 Lactobacillus (Lb.) brevis, 그리고 FGL0007은 Lactococcus (Lc.) lactis에 가장 유사한 것으로 확인하였다. Lb. brevis FSFL0004와 FSFL0005는 발효된 아귀로부터 분리되었고, Lb. brevis FSFL0036는 갈치 젓갈로부터 분리되었으며, Lc. lactis FGL0007 균주는 참가자미의 내장으로부터 분리되었다. 고속액체크로마토그래피(HPLC)를 사용한 정량분석결과를 보면, FSFL0004, FSFL0005, FSFL0036과 FGL0007에서 각각 10,754.37 μg/ml, 13,082.79 μg/ml, 12,290.19 μg/ml, 45.07 μg/ml의 GABA가 생성되었다. GABA가 풍부한 낙농제품의 발효 종균으로서 상용화 실험을 위해 1% MSG를 포함한 탈지방우유에 4개의 균주를 각각 접종하였다. TLC 결과를 보면 4개의 균주 모두가 GABA 생성을 보였다. HPLC 분석 결과를 보면, 4균주 중 Lc. lactis FGL0007이 가장 높은 GABA 생성(431.42 μg/ml)을 보였다. 본 연구의 내용은 GABA가 함유된 유제품 개발의 기반이 될 수 있을 것으로 생각한다. Lactic acid bacteria were isolated from a variety of fermented seafoods and sea creatures from the East Sea Rim, Korea and were screened for γ-amino butyric acid-producing (GABA) activity. Through a 16S rRNA sequence analysis, the bacteria of interest, which were GABA-positive on the thin-layer chromatography analysis, were recognized as three isolates of Lactobacillus (Lb.) brevis and one isolate of Lactococcus (Lc.) lactis. Lb. brevis FSFL0004 and FSFL0005 were isolated from fermented anglerfish and Lb. brevis FSFL0036 was derived from salted cutlass fish. The Lc. lactis strain FGL0007 was isolated from the gut of a brown sole flounder. According to HPLC analysis, the GABA contents produced by FSFL0004, FSFL0005, FSFL0036 and FGL0007 were equivalent to 10,754.37 μg/ml, 13,082.79 μg/ml, 12,290.19 μg/ml, and 45.07 μg/ml respectively in 1% monosodium glutamate-supplemented methionyltRNA synthetase (MRS) broth. The four strains were inoculated in skim milk with 1% monosodium glutamate to commercialize the strains as starter cultures for GABA-enriched dairy products, and TLC results displayed the production of γ-amino butyric acid by all four strains in the adaptation media. Lc. lactis FGL0007 demonstrated the greatest GABA production (431.42 μg/ml) by HPLC analysis. The GABA production by lactic acid bacteria strains in the skim milk demonstrated in the present study may be helpful for the production of GABA-enriched dairy products.

Bioluminescence 방법에 의한 우유 및 유제품에 존재하는 세균수 측정에 관한 연구

이승배,도형기 건국대학교 1986 論文集 Vol.22 No.1

Determination of bacterial ATP based on luciferin-luciferase bioluminescence reaction was applied to the measurement of bacteria in raw milk and liquid yoghurt as in alternative rapid method. The results are as follows : 1. The light yield was proportional to the concentration of ATP giving a straight light within a range of 10-10 to 10-6M. 2. The light yield related with the ATP content was increased as the number or bacteria in E. coli and L. bulgaricus was increased. When ATP content of bacteria were determined by bioluminescence reaction and compared. 3. The patterns of growth curve for E. coli look very similar, when they were monitored by ATP content assayed by bioluminescence reaction and total colony count. 4. The bacterial ATP of each sample in raw milk and liquid yoghurt was proportional to the number of bacteria, Coefficient of correction was 0.74 and 0.58 respectively. Thus, this new rapid method based on determination of bacterial ATP monitored by luciferin-luciferase bioluminescence reaction can be used for the indirect counting of bacteria, respectively.